A Decade of EU-funded Animal Health

KI-31-11-002-EN-C
A decade of EU-funded
This publication gathers information on initiatives
funded by the European Commission over the last
decade regarding animal health challenges. There are
Animal Health
A decade of EU-funded Animal Health Research

some general actions which have been developed for
the harmonisation of European funding schemes, for
the prioritization of research topics or for implementing
improved surveillance systems on emerging and re-
emerging diseases. Several projects concern notifiable
diseases, which are often highly epidemic and are
Research
frequently regulated by the culling of infected animals.
The goal pursued is to achieve the capacity to react more
quickly by using fast and reliable diagnostics tools, but
also to develop alternative disease eradication protocols
through new generation vaccines. Regarding endemic
diseases, which decrease the efficiency and profitability
of the livestock sector and are
difficult to phase out, the aim is to progressively reduce
their incidence. Finally, there is a chapter which gives
information on projects targeting the influenza viruses,
for controlling animal infection and improving our
knowledge on cross-species infection within a one
health vision.
Research and
Innovation
How to obtain EU publications
Free publications:
This publication is dedicated to the memory of Isabel Minguez
via EU Bookshop (http://bookshop.europa.eu);
Tudela who made a major contribution to the Animal Health at the European Unions representations or delegations. You can obtain their contact details on the
research sector over many years in the Commission services, but Internet (http://ec.europa.eu) or by sending a fax to +352 2929-42758.
who sadly passed away on 16 April 2011.
Priced publications:
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EUROPEAN COMMISSION
Directorate-General for Research and Innovation

Directorate E Biotechnologies, Agriculture, Food
Unit E4 Agriculture, Forest, Fisheries, Aquaculture
E-mail: RTD-FP7-KBBE-Activity1@ec.europa.eu
Contact: Anne-Sophie Lequarre
European Commission
Office SDME 08/018
B-1049 Brussels
Tel. (32-2) 29-80068

Fax (32-2) 29-63029
E-mail: Anne-Sophie.Lequarre@ec.europa.eu
EUROPEAN COMMISSION
Animal Health Research
FP7 Cooperation Work Programme

Theme: Food, Agriculture and Fisheries, and Biotechnology

2012 Biotechnologies, Agriculture, Food EN
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5
Contents
FOREWORD 9
CHAPTER 1. NETWORK, SURVEILLANCE 11
[EMIDA] Coordination of European research on emerging

and major infectious diseases of livestock 12
[STAR-IDAZ] Global Strategic Alliances for the Coordination of Research

on the Major Infectious Diseases of Animals and Zoonoses 17
[EPIZONE] Network of Excellence

for Epizootic Disease Diagnosis and Control 21
[DISCONTOOLS] Development of the most effective tools

to control infectious animal diseases 24
[Arbo-Zoonet] Network for Capacity-building

for the Control of Emerging Viral Vector-borne Zoonotic Diseases 27
[WildTech] Novel Technologies for Surveillance of Emerging

and Re-emerging Infections of Wildlife 31
[ICONZ] Integrated control of neglected zoonoses:

improving human health and animal production
through scientific innovation and public engagement 36
CHAPTER 2. EPIDEMIC DISEASES OF LIVESTOCK 41
European policies for animal health sustained

by EC-funded research project
Introduction by Jim Scudamore and Alberto Laddomada 42
2.1 Foot-and-mouth disease 44
[FMD_ImproCon] Improvement of foot-and-mouth disease control

by ethically acceptable methods based on scientifically validated assays
and new knowledge on FMD vaccines, including the impact of vaccination 44
[CA FMD/CSF] Foot-and-mouth disease (FMD)

and classical swine fever (CSF) coordination action 50
[FMD-Disconvac] Development, enhancement and complementation

of animal-sparing, foot-and-mouth disease vaccine-based control strategies
for free and endemic regions 54
6 A D E C A D E O F E U - F U N D E D A N I M A L H E A L T H R E S E A R C H
2.2. Classic swine fever 59
[CSFVACCINE & WILD BOAR] Epidemiology and control

of classical swine fever (CSF) in wild boar and potential use
of a newly developed live marker vaccine 59
[CSF-GoDIVA] Improve tools and strategies for the prevention

and control of classical swine fever 66
2.3. African swine fever 71
[ASFRISK] Evaluating and controlling the risk

of African swine fever in the EU 71
2.4. Orbivirus 77
[BTVAC] Improved vaccines for bluetongue disease 77
[Medreonet] Surveillance network of reoviruses, bluetongue

and African horse sickness, in the Mediterranean basin and Europe 82
[ORBIVAC] Development of vaccines for bluetongue virus (BTV),

epizootoc hemorrhagic disease virus (EHDV) and African horse sickness virus (AHSV) 89
CHAPTER 3. ENDEMIC DISEASES 95
The economic impact and research synergy resulting

from EC-funded projects
Introduction by Alex Morrow and Declan OBrien 96
3.1. Mycobacterial diseases 101
[VENoMYC] Veterinary network of laboratories researching

into improved diagnosis and epidemiology of mycobacterial diseases 101
[TB-STEP] Strategies for the eradication of bovine tuberculosis 107
3.2. Parasites 112
[DELIVER] Design of effective and sustainable control strategies

for liver fluke in Europe 112
[PARASOL] Novel solutions for the sustainable control

of nematodes in ruminants 117
[PARAVAC] Vaccines against helminth parasites of livestock

of economic and/or public health significance 121
3.3. Tick-borne diseases 125
[PiroVac] Improvement of current and development of new vaccines

for theileriosis and babesiosis of small ruminants 125
7
3.4. Porcine circovirus diseases 129
[PCVD] Control of porcine circovirus diseases (PCVD):

towards improved food quality and safety 129
[NMSACC-PCVD] PCVD: Towards improved food quality and safety

within EU new Member States (NMS) and associate candidate countries (ACC) 136
3.5. Porcine reproductive and respiratory syndrome 141
[PoRRSCon] New tools and approaches to control Porcine Reproductive

and Respiratory Syndrome (PRRS) in the EU and Asia 141
CHAPTER 4. INFLUENZA 147
Introduction by Ilaria Capua 148
4.1. Avian flu 150
[AIV-VACC] Vaccine, diagnostic test development

and immunology aspects of avian influenza 150
[FLUAID] Generation of information and tools to support the management

of the avian influenza crisis in poultry 155
[Novaduck] Novel avian influenza (AI) DIVA recombinant vaccines for duck 158
[Healthy Poultry] Development of new integrated strategies

for prevention, control, monitoring of epizootic poultry diseases 163
[FLUPATH] Avian influenza: Impact of virus-host interactions

on pathogenesis and ecology 166
[New-Flubird] Network for early warning of influenza viruses

in migratory Birds in Europe 172
[FLURESIST] Avian influenza virus survival in poultry commodities,

poultry manure and the environment 179
[Rivers] Resistance of influenza viruses in environmental reservoirs

and systems 182
[FLUTEST] Improved diagnosis and early warning systems

for avian influenza outbreak management 184
[FLU-LAB-NET] Development and enhancement

of laboratory networks for avian influenza 189
[FLUTRAIN] Training and technology transfer

of avian influenza diagnostics and disease management skills 195
[ConFluTech] Capacity-building for the control of avian influenza

through technology transfer and training 198
4.2. Swine flu 202
[ESNIP 2] European Surveillance Network for Influenza in Pigs 2 202
[ESNIP 3] European Surveillance Network for Influenza in Pigs 3 204
[FLUPIG] Pathogenesis and transmission of influenza in Pigs 211

9
Foreword
As we move towards a predicted global outbreak of foot and mouth disease in

population of 9 billion by 2050, with an the UK in 2001 may have been close to
expected 70% increase in world food 8 billion as a result of disruption in trade
demand and meat consumption projected and impacts on industries such as tour-
to double to 465 million tonnes, food secu- ism. Bovine Spongiform Encephalopathy is
rity and animal health are more important estimated to have cost the UK 8.5 billion
than ever. and more recently bluetongue in the Neth-
erlands was estimated to have cost 155
In February I announced the Commissions million. The costs related to endemic condi-
proposal for a European Bioeconomy Strat- tions such as mastitis or lameness, are also
egy, a key component of which strives to reaching several billion Euros per year for Mire GEOGHEGAN-QUINN
ensure that the importance of the primary the European dairy industry. Commissioner for
Research, Innovation
production sector is clearly recognised, and Science
reconciling demands for agriculture and The need to develop innovative and bet-
European Commission
food security with the sustainable use of ter tools to control disease is now more
resources, while ensuring environmental critical than ever. The lessons learnt from
protection. a decade of EU research are shaping the
new research and innovation programme
While primary food production is intensi- (Horizon 2020), which follows the cur-
fied to meet increasing demand, the result- rent programme in 2014, as food secu-
ing increased international trade leads to rity is covered by the societal challenge
greater movement of animals and animal food security, sustainable agriculture,
products and higher risk for animal health. marine and maritime research and the
Infectious diseases directly affect live- bio-economy.
stock production with consequences for
food security and food safety, trade, rural This catalogue brings together the fruits
development, and the environment, while of European research efforts targeting
also affecting the livelihood of farmers. animal health; it includes forty projects
Climate change is expected to influence with a total budget close to 100 mil-
the spread of certain pathogens; we have lion euros. The scope of the work is vast,
recently experienced the rapid appear- ranging from new vaccine development,
ance across Europe of new diseases such enhanced epidemiological models, better
as those caused by blue tongue or Schmal- surveillance activities and improved diag-
lenberg viruses. In addition animal health is nostics methods. These research results
also important in preventing the spread of help the development of European policies
certain potentially dangerous diseases from to better manage disease surveillance,
animals to humans. prevention, control and eradication. There
have been several outstanding successes
Animal disease can have devastating socio- particularly in developing tools for the
economic effects: in Europe for example effective control of swine diseases such
the total cost to the rural economy of the as porcine coronavirus disease or classic
swine fever as well as enhancing European

expertise on the avian and swine influenza
viruses, knowledge which has been widely
deployed by industry.
This publication highlights the impact of our

research funding in guaranteeing the over-
all health and welfare of European livestock
as well as contributing to the high quality
and performance of this sector. I am con-
vinced that new innovation opportunities
will arise from sharing this knowledge more
widely.
CHAPTER 1.
Network,
surveillance
[EMIDA]
Coordination of
European research on
emerging and major infectious
diseases of livestock
Acronym: Summary activities and working towards a common
EMIDA The disease threats to the livestock industry research agenda and mutual research
have increased steadily over the past decades funding activities.
Project number:
219235 as a result of globalisation, evolving patho-
gens and climate change. Responding to ani- The aim of the EMIDA ERA-NET is to build on
EC contribution:
mal disease threats relies heavily on science; and accelerate the work of the SCAR CWG
EUR 997 218
research makes a significant contribution to in the field of animal health. The scope of
Duration: the development of disease control policy the project includes emerging and major
45 months
and the translation of policy, and other driv- infectious diseases of production animals,
Start date: ers for improving animal health, into practical including fish and bees and including those
1 April 2008 effect. Although the legislation that underpins conditions which pose a threat to human
Instrument: policy for the control of statutory diseases is health but excluding food safety issues
Coordination determined at the EU level, the research that relating to the handling of livestock products
and support action supports policy development and implemen- and diseases of wildlife except where they
tation is primarily carried out at the national act as reservoirs of infection for humans or
level and is largely uncoordinated as is the production animals.
research on other major infectious diseases
currently affecting livestock production. The objectives of the ERA-NET are being
delivered through the following four work
Improved coordination and collaboration packages: WP1: Project coordination, man-
of this research activity is therefore vital agement, communication and dissemin
to ensure the efficient and effective under- ation; WP2: Mapping and analysis of existing
pinning of EU and national policy and the research and current needs and information
sustainability of the European livestock on the commissioning and management of
and animal health industries and the ani- joint programmes; WP3: Develop, test, evalu-
mal health science capacity. A Collaborative ate and refine instruments (Pilots) and WP4:
Working Group (CWG) on animal health and Developing a strategic transnational animal
welfare under the Standing Committee of health research agenda.
Agriculture Research (SCAR) was developed
to address this gap, with the objectives of
developing a durable, focused network of Problem
national research funders in the EU Member Most of the funding for research on animal
States and associated countries for the pur- health in Europe comes from the EU Member
pose of sharing information, coordinating States and associated countries, involving
C H A P T E R 1 . N E T W O R K , S U R V E I L L A N C E 13
government ministries (agriculture and for both EMIDA and the CWG. Details of
health) or research councils. All these fund- over 2 000 projects have been uploaded
ing bodies have their own agendas/priorities to the project database by the project
that are set independently of each other thus partners. To complete the mapping of the
resulting in fragmentation and potentially research landscape, three other databases
duplication of effort. of supporting information in the form of
publications over the past four years, inter-
national animal health-related patents and
Aim EC-funded animal health-related projects
To build on, and accelerate the work of, the have also been developed using data from
SCAR CWG in developing a durable, focused international scientific databases. These
network of national research funders in the databases, the methodology behind them
EU Member States and associated coun- and the associated reports on research
tries for the purpose of sharing information, outputs can be accessed on the project
coordinating activities and working towards website. A questionnaire survey on cur-
a common research agenda and mutual rent management practices relating to the
research funding activities in the field of research programmes of the project part-
animal health. ners and their perceived needs and priority
topics of interest for inclusion in a common
call was conducted and the resulting report
Project activities is also available on the project website.
EMIDA started on 1 April 2008 and cur-
rently has 29 partners from 19 countries Development and pilot of
(15 Member States, Israel, Norway, Switzer- instruments for common calls
land and Turkey) with a combined annual Through the aforementioned question-
research budget for work on animal health naire, a large number of topics were sug-
in the region of EUR 270 million. There are gested as possible subjects for a common
also four associated partners. The project research call. A matrix approach based on
partners consist of ministries of agriculture, disease groups and technologies was used
ministries of health/public health, ministries to rationalise these and from this, four
of education/research councils and agen- broad topics were identified and agreed at
cies under the aforementioned. The varied the second Project Consortium meeting.
background of the organisations involved, Analysis of partners programme manage-
each with their own agendas/priorities, ment practices provided the basis for those
encompasses the funding of research practices and instruments employed in the
across the spectrum from basic to strategic EMIDA common call. It was agreed that a
and applied science. virtual common pot funding mechanism
would be utilised. The pilot common call,
Systematic exchange with a budget in the region of EUR 20 mil-
of information lion, based on the following four topics was
A project website has been established opened in early September 2009.
which is linked to the CWG website and
associated project database. The EMIDA Vector-borne diseases Development
website contains a password-protected dis- of underpinning knowledge and tools for
cussion forum allowing exchange of infor- early warning, detection and monitoring
mation between project partners. A frame- and novel control strategies.
work was established under the CWG for Zoonoses and antimicrobial resistance,
the capture of research project information excluding microbial safety of prod-
and a database developed which is serving ucts Development of underpinning
the needs for the collection of information knowledge and tools for early warning,
detection and monitoring and novel Project outputs achieved so far

control strategies. The major project outputs to date are as
Major infectious diseases affecting pro- follows (links):
duction Development of underpinning
knowledge and tools for early warning, Project website: http://www.emida-era.net
detection and monitoring and novel Methodology For EMIDA publications
control strategies, including genetics of database
resistance. report on the mapping of European
Aquaculture Development of under- research publications on infectious
pinning knowledge and tools for early diseases of livestock
warning, detection and monitoring and Repor t on mapped and analysed
novel control strategies, particularly data and information from National
vaccine-based approaches. Programmes FPU Terms of Reference
Overview of foresight studies
Twelve projects to a total value of over Pilot Research Call with an online sub-
EUR21 million have been funded, two mission system launched on 7 Septem-
on Topic 1 (Vector-borne diseases), two ber 2009 http://www.submission-emida-
on Topic 2 (Zoonoses and antimicrobial era.net
resistance, seven on Topic 3 (Major infec- Second Common Call with a budget of
tious diseases affecting production) and EUR 20 million launched on 7 March 2011
one on Topic 4 (Diseases of fish in aqua-
culture). Details of these projects can be Expected final results and their
found online (http://www.emida-era.net/ potential applications
index.php?page=content&id=17). A second and impact
common call was launched in March 2011 A sustainable structure will be established
with a budget in the region of EUR 20 mil- for the collaborative working group with
lion with the proposal expected to address sharing of information at all levels, and tak-
one of seven activity lines or the associated ing forward a common research agenda
specific topics. developed by the Foresight and Program-
ming Unit using procurement procedures
Strategic research agenda developed and piloted under EMIDA. This will
development reduce duplication of effort and focus the
A Foresight and Programming Unit (FPU) was available funding on addressing the research
established which identified and reviewed needs for improved control of emerging and
all the relevant foresight studies, identifying major infectious diseases of livestock.
issues and drivers and a report has been pro-
duced which is available on the project web-
site. This work was further developed and Project website
refined in a Delphi study, for which partici- http://www.emida-era.net
pants from a broad range of disciplines were
engaged and, following a consensus work- Keywords
shop, a draft long-term strategic research infectious diseases of animals, research
agenda was produced. Terms of reference for coordination, animal health
the future of the FPU beyond the end of the
EMIDA ERA-NET have been developed and Coordinator
are available on the project website. These Dr Alex Morrow
will be incorporated into the collaborative Department for the Environment,
agreement which is currently being devel- Food and Rural Affairs (Defra)
oped, outlining a sustainable structure for UNITED KINGDOM
the collaborative working group. Alex.Morrow@defra.gsi.gov.uk
Partners
ORGANISATION COUNTRY CONTACT E-MAIL
Institut national FRANCE Dr Thierry Pineau Thierry.Pineau@toulouse.inra.fr
de la recherche
agronomique (INRA)
Federal Ministry GERMANY Dr Claudia Herok Claudia.Herok@bmbf.bund.de
of Research and
Education (BMBF)
Forschungszentrum GERMANY Dr Petra Schulte Petra.Schulte@fz-juelich.de
Jlich, Project
Management Jlich
(FZJ-PTJ)
Ministry of Economic NETHERLANDS Dr Albert Meijering A.Meijering@minlnv.nl
Affairs, Agriculture
and Innovation
(EL&I)
Food and Consumer NETHERLANDS Dr Wim Ooms Wim.Ooms@vwa.nl
Product Safety
Authority (VWA)
Ministero del Lavoro, ITALY Dr Marina Bagni Marina.Bagni@sanita.it
della Salute e delle
Politiche Sociali
(HM-DVPHNFS)
Ministry of Agri- ITALY Dr Alberto Masci A.Masci@politicheagricole.it
cultural Food and
Forestry Policies
(MiPAAF)
Ministry of Food, DENMARK Dr Michael Knudsen Mikn@dffe.dk
Agriculture and Fish- (DFIA)
eries, Danish Food
Industry Agency
(DFIA)
Ministry of Agricul- CZECH Dr Milan Podsednicek Milan.Podsednicek@mze.cz
ture, Department of REPUBLIC
Research, Education
and Advisory Services
(MZE)
Federal Ministry of AUSTRIA Dr Hermann Hermann.Schobesberger@
Health (BMG) Schobesberger vetmeduni.ac.at
(AGES)
Belgian Federal BELGIUM Dr Dominique Dominique.Vandekerchove@
Public Service of Vandekerchove health.fgov.be
Health, Food Chain
Safety and Environ-
ment, Service of
Contractual Research
(FPS-CR)
Federal Agency for BELGIUM Dr Xavier Van Huffel Xavier.VanHuffel@favv.be
the Safety of the
Food Chain, Belgium
(FASFC)

Veterinary and Agro- BELGIUM Dr Hein Imberechts Hein.Imberechts@var.fgov.be
chemical Research
Centre (CODA-CERVA)
Ministry of Agricul- FINLAND Dr Katri Levonen Katri.Levonen@mmm.fi
ture and Forestry
(MMM)
Department of Agri- IRELAND Dr John Egan John.Egan@agriculture.gov.ie
culture, Fisheries and
Food (DAFF)
Research Council of NORWAY Dr ystein Rnning oro@forskningsradet.no
Norway (RCN)
Swedish Research SWEDEN Dr Johanna Dernfalk Johanna.Dernfalk@formas.se
Council for Environ-
ment, Agricultural
Sciences and Spatial
Planning (Formas)
Biotechnology and UNITED Dr Sadhana Sharma Sadhana.Sharma@bbsrc.ac.uk
Biological Sciences KINGDOM
Research Council
(BBSRC)
Swiss Federal SWITZERLAND Dr Irene Schiller Irene.Schiller@bvet.admin.ch
Veterinary Office
(SFVO)
Scottish Government UNITED Dr Sarah Gledhill Sarah.Gledhill@scotland.gsi.
(SG) KINGDOM gov.uk
Ministry of Agri- CYPRUS Dr Penelope pstylianou@vs.moa.gov.cy
culture, Natural Stylianou
Resources and Envir
onment of Cyprus
(Veterinary Services)
(VS)
The Ministry of Agri- LITHUANIA Prof. Antanas antanas@lva.lt
culture of Lithuania Sederevicius
(MAL)
Ministry of Agri- ISRAEL Dr Boris Yakobson dir-kimron@moag.gov.il
culture and Rural
Development (Israeli
Veterinary Services
and Animal Health)
(IVSAH)
Ministry of Agricul- TURKEY Dr Erhan Bilge ebilge@tagem.gov.tr
ture and Rural Affairs
(General Director
ate of Agricultural
Research) (TAGEM)
National Institute SPAIN Ms Anabel de la Pea Anaisabel.delapena@inia.es
for Agriculture and
Food Research and
Technology (INIA)
French National FRANCE Dr Serawit Bruck Serawit.Bruck@agencere-
Research Agency cherche.fr
(ANR)

Federal Agency for GERMANY Dr Elke Saggau Elke.Saggau@ble.de
Agriculture and Food
(BLE)
Federal Ministry of GERMANY Dr Hartmut Stalb Hartmut.Stalb@bmelv.bund.de
Food, Agriculture and
Consumer Protection
(BMELV)
[STAR-IDAZ]
Global Strategic Alliances for

the Coordination of Research on
the Major Infectious Diseases
of Animals and Zoonoses
Summary groundwork established by the SCAR Col- Acronym:
Animal diseases can cause serious social, laborative working group on animal health STAR-IDAZ
economic and environmental damage and welfare research, the EMIDA ERA-NET
Project number:
and, in some cases, also threaten human project and specific INCO-NETs involving 265919
health. An increasing number of the major partner countries. The scope of the project
EC contribution:
disease problems or threats faced by the includes coordination of research relevant
EUR 999 130
livestock industry and zoonoses are of a to emerging and major infectious diseases
global nature. The overall aim of the global of livestock, including fish and managed Duration:
48 months
strategic alliances for the coordination of bees, and those infections of livestock
research on the major infectious diseases that may carry the risk of disease threat Start date:
of animals is to improve coordination of to human health. Diseases of wildlife will 1 February 2011
research activities on the major infectious also be considered where they are identi- Instrument:
diseases of livestock and zoonoses so as fied as reservoirs of infection with emerging Coordination
and support action
to hasten the delivery of improved control and major infectious diseases of humans or
methods. This will be achieved through the production animals.
establishment of an international forum of
R & D programme owners/managers and These objectives will be delivered through
international organisations for the purpose the following five work packages: WP1:
of sharing information, improving collabo- Project coordination, management, com-
ration on research activities and working munication and dissemination; WP2: Shar-
towards common research agendas and ing information on existing research pro-
coordinated research funding on the major grammes; WP3: Analysis of and responding
animal diseases affecting livestock produc- to global, regional and industry sector
tion and/or human health. It will build on the priorities; WP4: Networking of ongoing
research activities on major issues; and develop durable procedures for a better
WP5: Developing a strategic transnational coordinated, rapid response to urgent
animal health research agenda. Regional research needs;
networks for (i) the Americas and (ii) Asia identify unique regions with localised
and Australasia are being established and diseases and improve access to research
it is hoped that aregional network covering in those areas;
the Middle East and Africa will follow. improve access to, and the use of
research results across all partner
organisations;
Problem facilitate the establishment of research
An increasing number of the major disease management capacit y and pro -
problems or threats faced by the livestock grammes in those partner countries
industry and zoonoses are of a global wishing to develop research activities
nature. Recent disease threats, such as the in this area.
global threat from H5N1 avian influenza,
H1N1 swine influenza and the spread of Expected results
bluetongue in Europe, highlight the need Specific project outputs should include a web-
for rapid coordinated research to provide based hub for information exchange, a web-
the evidence needed for the development based discussion forum, a long-term common
of effective control policies. Lack of coord strategic research agenda, collaboration on
ination between the funding bodies interna- the research activities relating to a number
tionally can result in duplication of effort in of priority diseases, including networking the
some areas and insufficient attention and research communities concerned, and coordi-
funding being given to other areas. This nation of new research requirements.
necessitates seeking wider coordination
and collaboration, if value for money and The major expected outcome is a long-term
rapid progress on disease control is to be sustainable network allowing exchange of
achieved. information and coordination of research
activities so as to hasten the development
of improved control methods.
Aim
The specific objectives of the global Potential benefits
network are to: Efficient deployment of national funds
for both national and transnational (joint)
strengthen the linkages between, and research, including research procurement
reduce the duplication of, global research in response to emergency situations
efforts on high priority infectious dis- Improved cost-effectiveness of commis-
eases of animals (including zoonoses); sioned research, by creating a consen-
maximise the efficient use of expertise sus on the level of funding that should
and resources; and accelerate coordi- be directed at given priorities of both
nated development of control methods; nationally and internationally funded
identify and coordinate the response to programmes
gaps in research activities for targeted Improved coordination of research pri-
diseases; orities suitable for future programme
create the necessary critical mass and funding
capacity to address emerging infectious Improved availability of validated and
disease threats; relevant research data for animal health
improve the cost-effectiveness and policymakers, and the animal health
added value to network partners of and livestock industries (including
current research programmes; aquaculture)
Improved availability of information on Keywords

national research capacity, including infectious diseases of animals, zoonoses,
expertise, in the various areas research coordination, animal health, global
Identification of the needs for building network
capacity and capability in animal health
research Coordinator
Improved consultation with other inter- Dr Alex Morrow
national policymakers and animal health Department for the Environment,
organisations (e.g. EFSA, ECDC, OIE, FAO, Food and Rural Affairs (Defra)
and WHO) UNITED KINGDOM
Alex.Morrow@defra.gsi.gov.uk
Project website
http://www.star-idaz.net
Partners
Chinese Academy of CHINA Dr Yi Zhang zhangyi@mail.caas.net.cn
Agricultural Sciences
(CAAS)
Institut national FRANCE Dr Thierry Pineau Thierry.Pineau@toulouse.inra.fr
de la recherche
agronomique (INRA)
Empresa Brasileira BRAZIL Janice Ciacci Zanella Janice@cnpsa.embrapa.br
de Pesquisa
Agropecuria
(Embrapa)
Ministry of Science NEW ZEALAND Dr Prue Williams Prue.Williams@msi.govt.nz
and Innovation (MSI)
Ministerio de Ciencia, ARGENTINA Dr Eduardo Trigo ejtrigo@gmail.com
Tecnologa e Inno-
vacin Productiva
(MINCYT)
United States UNITED Dr Cyril Gay Cyril.Gay@ars.usda.gov
Department STATES
of Agriculture
(USDA-ARS)
Consejo Tcnico MEXICO Prof. Juan Garza jgarza@unam.mx
Consultivo Nacional
De Sanidad Animal
(CONASA)
Ministry of Food, DENMARK Dr Michael Knudsen Mikn@dffe.dk
Agriculture and
Fisheries, Danish
Food Industry Agency
(DFIA)
Forschungszentrum GERMANY Dr Petra Schulte Petra.Schulte@fz-juelich.de
Jlich, Project
Management Jlich
(FZJ-PTJ)

Canadian Food CANADA Dr Primal Silva Primal.Silva@inspection.gc.ca
Inspection Agency
(CFIA)
Moscow State Acad- RUSSIAN Prof. Larisa lag22004@mail.ru
emy of Veterinary FEDERATION Gnezdilova
Medicine and Bio-
technology named
after K.I. Skryabin
(MGAVM and B)
International Centre RUSSIAN Irina Kuklina kuklina@mniop.ru
for Innovations in FEDERATION
Science, Technol-
ogy and Education
(ICISTE)
Ministry of Economic NETHERLANDS Dr Albert Meijering A.Meijering@minlnv.nl
Affairs, Agriculture
and Innovation
(EL&I)
Ministero della ITALY Dr Marina.Bagni Marina.Bagni@sanita.it
Salute (DVPHNFS)
National Institute SPAIN Dr Nuria Duran Vila Nuria.Duran@inia.es
for Agriculture and
Food Research and
Technology (INIA)
Pfizer International FRANCE Dr Theo Kanellos Theo.Kanellos@pfizer.com
Operations (PAH)
Department of Agri- AUSTRALIA Dr Mike Nunn Mike.Nunn@daff.gov.au
culture, Fisheries and
Forestry (AG DAFF)
Ministry of Science INDIA Dr Shailja Vaidya shailja@dbt.nic.in
and Technology Gupta
(DBT)
Merial SAS (MAH) FRANCE Dr Jules Minke Jules.Minke@merial.com
Biotechnology and UNITED Dr Sadhana Sharma Sadhana.Sharma@bbsrc.ac.uk
Biological Sciences KINGDOM
Research Council
(BBSRC)
International Fed- BELGIUM Barbara Freischem B.Freischem@ifahsec.org
eration for Animal
Health (IFAH)
Associated partners
Wellcome Trust UNITED Dr Lara Bethke L.Bethke@wellcome.ac.uk
KINGDOM
The African Union AFRICAN Dr Baba Soumare Baba.Soumare@au-ibar.org
Interafrican Bureau UNION
for Animal Resources
(AU-IBAR)

Agriculture, Forestry JAPAN Dr Satoshi Shiku Satoshi_Shiku@nm.maff.go.jp
and Fisheries
Research Council
Secretariat (AFFRC)
European Food EUROPEAN Dr Per Have Per.Have@efsa.europa.eu
Safety Authority UNION
(EFSA)
World Organisation INTERNATIONAL Dr Elisabeth E.Erlacher-Vindel@oie.int
for Animal Health Erlacher-Vindel
(OIE)
[EPIZONE]
Network of Excellence
for Epizootic Disease
Diagnosis and Control
Summary improvement of excellence through Acronym:
EPIZONE brings together expert scientists cooperation. EPIZONE
to combine and harmonise efforts on the
Project number:
development of new strategies and tools EPIZONE has developed a network of more 016236
to combat future epizootic animal diseases. than 350 key scientists with an inter
EC contribution:
Epizootic diseases in food-producing animals national reputation, complementary exper-
EUR 14 000 000
including aquaculture constitute a major risk tise and skills working together within
for food safety and food security and there- Europe and worldwide. Through increased Duration:
60 months
fore have become a concern for both animal excellence by collaboration of veterinary
and human health. Such diseases spread very institutes from Europe, China and Turkey, Start date:
fast in high densities of susceptible animals the economic and social impact and the 1 June 2006
through animals, vectors or animal products. public health risk of future outbreaks of Instrument:
Outbreaks in Europe have had enormous foot-and-mouth disease, classical swine Network
social and economic impact, and need to fever, avian influenza and other relevant of Excellence
be addressed across the whole production epizootic disease like bluetongue and Afri-
chain of animal-related food. To develop new can swine fever, will be reduced.
methods for the prevention and control of
epizootic diseases, collaborations of scientific EPIZONE has been developed for the integra-
institutes are indispensable and international tion of scientists in health and production of
harmonisation and standardisation of proce- animals at the European level. The benefits
dures is urgently needed. of EPIZONE primarily concern consumers
and stakeholders throughout the food sup-
The objective of EPIZONE is to improve ply chain but also agriculture administra-
research on preparedness, prevention, tions and biotechnology companies. EPIZONE
detection and control of epizootics by includes 18 institutes from 12 countries.
Partners maintain networks worldwide, detection, and control of epizootic animal

linked to EPIZONE, and most have (inter) diseases through cooperation, with extra
national reference lab-based tasks for con- attention for new and emerging epizootic
trol of epizootics. The management structure animal diseases including these which may
of EPIZONE generates durable interactions have zoonotic potential. This includes the
among partners. EPIZONE includes the FAO goal to reduce the economic and social
as a world-oriented organisation, and also an impact of future outbreaks of foot-and-
enterprise (SME) specialised in dissemination mouth disease, classical swine fever, avian
of knowledge via the Internet. Within organ influenza (AI) and other relevant epizootic
isational work packages integration activi- diseases such as bluetongue and African
ties, including communications, meetings swine fever, through increased excellence
and trainings have been developed. Within by collaboration.
scientific work packages, joint research is
executed covering four thematic areas: diag-
nostics, intervention strategies, surveillance, Results
epidemiology and risk assessment. Given the EPIZONE has established an international
network structure, the technical resources network for sharing knowledge and expert
and the scientific excellence, EPIZONE ise on epizootic animal diseases and mater
assures strategically driven state-of-the-art ials and reagents. Increased excellence by
research of world-renowned quality. collaboration generated by the project will
help to combat future epizootic animal
disease more efficiently and more cost-
Problem effectively. Improved knowledge on epizo-
In recent years, much time and money has otic disease diagnosis, epidemiology, risks
been devoted to fighting and controlling out- and intervention strategies, generated by
breaks of well-known major animal diseases, EPIZONE will contribute to the prevention
such as avian influenza, classical swine fever and control of epizootic animal diseases on
and foot-and-mouth disease. The threat a global level. In this way, European experi
of new emerging and re-emerging epizo- ence, knowledge and scientific achieve-
otic animal diseases seems to increase in ments will gain visibility and will be used
all parts of the world. Epizootic diseases to support decision-making and future
constitute a major risk to food production. research.
Such diseases spread very quickly through
animals, vectors or animal products espe- EPIZONE has achieved the following.
cially where susceptible animals are kept
at a high density. Outbreaks showed enor- Created strong collaboration between
mous social and economic impact, and need veterinary institutes and with non-EU
to be addressed across the whole produc- partners through staff exchanges, sci-
tion chain of animal-related food. Animal entific missions, workshops, training and
disease control methods involving the mass technology transfer, and also bringing
culling of livestock are no longer acceptable together next-generation scientists. After
for the international society. Innovative and its EU-funded period, which ends 1 Janu-
rapid prevention and control strategies will ary 2012, EPIZONE will continue as the
be needed and international cooperation EPIZONE European Research Group.
becomes more and more important. Established online databases on refer-
ence materials, diagnostic protocols, cell
cultures, viruses, and disease outbreaks.
Aim Accommodated the development, val
The mission of EPIZONE is: To improve idation and harmonisation of detection
research on preparedness, prevention, methods including new molecular
techniques, pathogenic virus-specific Project website

diagnostics and pen-side tests, using http://www.epizone-eu.net
test panels generated by partner
institutes. Keywords
Provided important data on epidemi- epizootic diseases, animal diseases control,
ology, surveillance and risks of recent network of excellence
epizootic animal disease outbreaks in
the EU including bluetongue and avian Coordinator
influenza. Dr Wim H. M. van der Poel
Produced novel reagents and new Central Veterinary Institute
methods for development of new vac- Lelystad
cine technologies. Department of Virology
Generated scientific publications in P.O. Box 2004
international peer-reviewed journals. 8203 AA Lelystad
NETHERLANDS
wim.vanderpoel@wur.nl
Partners
ORGANISATION COUNTRY CONTACT
Friedrich-Loeffler-Institut GERMANY Dr Martin Beer
Greifswald-Insel Riems
Institute for Animal Health UNITED KINGDOM Dr Linda Dixon
Pirbright
Veterinary Laboratories Agency, UNITED KINGDOM Dr Anthony Fooks
Addlestone
Agence Franaise de Scurit Sanitaire FRANCE Dr Philippe Vannier
des Aliments
Maisons-Alfort
Danish Institute for Food DENMARK Prof. Sren Alexandersen
and Veterinary Research
Kalvehave
Statens Veterinarmedicinska Anstalt SWEDEN Dr Ulla Carlsson
Uppsala
Centre de coopration internationale FRANCE Dr Emmanuel Albina
en recherche agronomique pour
le dveloppement
Montpellier
Center for Animal Health SPAIN Dr Marisa Arias
Valdeolmos
Istituto Zooprofilattico Sperimentale ITALY Dr Ilaria Capua
delle Venezie
Legnaro
Lanzhou Veterinary Research CHINA Dr Yin Hong
Institute CAAS
Gansu
National Veterinary Research Institute POLAND Prof. Zygmunt Pejsak
Pulawy al Partyzantow
FMD Institute Ankara TURKEY Dr Fuat zyrk
Ankara
ORGANISATION COUNTRY CONTACT

Centrum voor onderzoek in BELGIUM Dr Frank Koenen
Diergeneeskunde en Agrochemie
Ukkel
University of Veterinary Medicine GERMANY Prof. Volker Moennig
Hannover
Istituto Zooprofilattico Sperimentale ITALY Dr Sylvia Bellini
della Lombardia e dellEmilia Romagna
Brescia
Harbin Veterinary Research CHINA Dr Hualan Chen
Institute CAAS
Harbin
Food and Agriculture Organisation ITALY Dr Keith Sumption
of the United Nations
Rome
Digital Value Internet Professionals NETHERLANDS Peter Oude Ophuis
Wageningen
[DISCONTOOLS]
Development of the most

effective tools to control
infectious animal diseases
Acronym: Summary will explore how new technologies can be
DISCONTOOLS DISCONTOOLS, an ongoing EU-funded pro- deployed more efficiently in the animal
ject, has three objectives. Firstly, to develop health research area.
Project number:
211316 a disease prioritisation methodology ena-
bling the prioritisation of research in order
EC contribution:
EUR 978 660
to stimulate the delivery of new or improved Problem
diagnostics, vaccines or pharmaceuticals. The concept for this proposal arose from
Duration: This will help to improve our ability to effec- of the work of the European Technology
48 months
tively control animal diseases which is a Platform for Global Animal Health (ETP-
Start date: key input into meeting the challenges of GAH) which was launched in December
1 March 2008 future food supplies. Once this methodol- 2004. Since then, the ETPGAH has devel-
Instrument: ogy is agreed with stakeholders, the objec- oped a vision, a strategic research agenda
FP7 Support tive is to establish a reference database (SRA) and an action plan to implement the
actions ensuring a clear focus on priority research recommendations in the SRA.
areas leading to more rapid breakthroughs
in technology development. Secondly, to Recent disease outbreaks have highlighted
develop a gap analysis for each of the pri- the necessity for not only producing new
oritised diseases to identify where research vaccines but also for improving existing
is needed. Thirdly, the DISCONTOOLS project vaccines and providing marker vaccines.
Effective tools for controlling animal dis- to ensure ma ximum returns for the
eases of major social and economic import investment in research.
ance are vital not only for Europe but also
for the rest of the world. The use of vac-
cines and diagnostic tests are a key compo- Aim
nent as they have the potential to support DISCONTOOLS will be carried out over four
control and eradication and to be highly years. It has three complementary strands
cost-effective. At present, there are no anti- for addressing the main objectives of the
viral medicines for use against the major FP7-KBBE-2007-1-3-03 call. These strands
viral diseases of animals. Consequently, all contribute to the primary objective of
vaccines and diagnostic tools are often the the call which is to enable research to be
only solution available for control. optimised by public and private funders
in a more effective manner to enable
New and improved vaccines are required new and improved tools to be developed
for a range of major animal diseases. In and delivered for the control of the major
addition, improved diagnostic tests must infectious diseases of animals including
be developed to enable the early diagno- zoonoses.
sis and detection of outbreaks along with
tests to demonstrate the effectiveness of
control programmes. The development of Expected results
new pharmacological or biocidal solutions The first strand will provide a validated
to the containment and control of disease database and peer-reviewed methodology
outbreaks also needs to be considered. in order to prioritise infectious animal dis-
eases. Gap analysis is the second strand
All these factors underline the need for a and will be carried out to identify those
coordinated, transparent and multidiscip areas where information and knowledge of
linary R & D effort from basic sciences the disease is deficient and where current
through to the emerging technologies and tools are lacking, inadequate or could be
on to product development, production, improved. Information will be collected in
authorisation and distribution. There is an a standard format for validation and entry
urgent need to boost research with effective into a specific disease database. A detailed
funding so that new or improved veterinary analysis will then be carried out for each of
medicines vaccines, pharmaceuticals the priority diseases to identify gaps in key
and diagnostic tests can be delivered. areas.
It is important to develop through pub- The third strand is to identify current and
lic and private partnerships an overview new technological tools that may be used
of current research and identify the gaps. to improve the ability to control infectious
Programmes can then be developed to fill animal diseases. The work will include
these gaps whilst at the same time devel- review of existing arrangements by stake-
oping research collaboration and syner- holders and the development of methodolo
gies to avoid duplication of research effort. gies to identify and evaluate new technol-
Within the EU, the lack of a formal mechan ogy. Effective identification and technology
ism to identify research gaps increases transfer is essential if new tools for disease
the reliance placed on scientific communi- control are to be developed.
ties, panels and workshops to assess these
needs. Assessments are limited and need One of the main features of the project is
continuous updating. It is equally important the involvement of a wide range of stake-
to adopt a global approach to ensure that holders who will actively participate in
research is coordinated and rationalised the governance of the project. This will
ensure that the stakeholders involved from of animal diseases, driven by stakehold-
research through to delivery of new con- ers, that will facilitate public and private
trol tools will be able to contribute to the funders in determining the most efficient
project. Dissemination of information from way of deploying research funding.
all three strands of work will be essential
if the project is to be successful. This will
be achieved through the communication References/publications
strategy which will include interactive web Brochure (http://www.discontools.eu/
systems and databases as an integral part documents/1380_DISCONTOOLSbro-
of the project. chure.pdf).
DISCONTOOLS methodology is available
To date, 12 sets of disease information on the website under Working Groups/
have been placed on the public website Documents of interest.
(http://www.discontools.eu). Expert groups DISCONTOOLS interactive disease data-
have completed their work in relation to base (http://www.discontools.eu/home/
an additional 18 diseases and this infor- disease_home).
mation is going through the approval pro-
cess. It is anticipated that the majority of
expert groups will have completed their Project website
work by the end of June 2011. In examin- http://www.discontools.eu
ing the results, it has become clear that the
information is quite complex and so we are Coordinator
developing an interpretation guide that will Declan OBrien
be placed on the website to clarify the use IFAH-Europe
of the results. In addition, each disease will Rue Defacqz 1
be accompanied by a short summary of 1000 Brussels
the main findings assisting all concerned in BELGIUM
interpreting the data. d.obrien@ifahsec.org
As the database is populated with the 51

diseases and as we receive input through
the public website, the data on the web-
site will become quite definitive in terms
of the state of play concerning research
prioritisation focused on delivering new
and improved diagnostics, vaccines and
pharmaceuticals. In addition, as the model
matures, we can then move on to a new
phase of adding further diseases. Via the
ongoing consultation process, the model
will remain up to date and prioritisation
will change over time as gaps are filled
and as new information becomes available
including on emerging and re-emerging
diseases.
Potential applications
Our wish is to create a definitive source of
data on prioritisation of research in the field
[Arbo-Zoonet]
Network for Capacity-building

for the Control of Emerging
Viral Vector-Borne Zoonotic
Diseases
Summary manner that includes vector control, vac-
Acronym:
Arboviruses are arthropod-borne viruses, cination programmes, improved therapy
Arbo-Zoonet
which include West Nile fever virus (WNFV), strategies, diagnostic tools and surveil-
a mosquito-borne virus, Rift Valley fever lance, public awareness, capacity-build- Project number:
211757
virus (RVFV), a mosquito-borne virus, and ing and improvement of infrastructure in
Crimean-Congo hemorrhagic fever virus endemic regions. EC contribution:
(CCHFV), a tick-borne virus. These arthro- EUR 998 470
pod-borne viruses can cause disease in Duration:
different domestic and wild animals and Problem 36 months
in humans, posing a threat to public health The virgin soil-epidemic raises the threat Start date:
because of their epidemic and zoonotic of expansion into other parts of Asia and 1 May 2009
potential. In recent decades, the geo- Europe. The general public concern regard- Instrument:
graphical distribution of these diseases ing emerging zoonotic disease has gained coordination
has expanded. Outbreaks of WNFV have interest and relevance in light of global and support action
already occurred in Europe, especially in warming. This is especially true regarding
the Mediterranean basin. Moreover, CCHFV the spread of the arboviruses such as RVFV,
is endemic in many European countries and CCHFV and WNFV, which are transmit-
serious outbreaks have occurred, particu- ted by mosquitoes or ticks. It is therefore
larly in the Balkans, Turkey and Southern imperative to work out integrated control
Federal Districts of Russia. In 2000, RVFV measures that include vector control, and
was reported for the first time outside the vaccination programmes, which improve
African continent, with cases being con- therapeutic strategies, as well as diagnos-
firmed in Saudi Arabia and Yemen. This tic tools and surveillance, public awareness,
spread was probably caused by ruminant capacity-building and the infrastructures in
trade and highlights that there is a threat endemic regions.
of expansion of the virus into other parts
of Asia and Europe. In the light of global
warming and globalisation of trade and Aim
travel, public interest in emerging zoonotic The Arbo-Zoonet project aims at creat-
diseases has increased. This is especially ing common knowledge of these diseases,
evident regarding the geographical spread as well as sharing and exchanging data,
of vector-borne diseases. A multidisci- expertise, experiences and scientific infor-
plinary approach is now imperative, and mation. The surveillance systems will be
groups need to collaborate in an integrated maintained and expanded, monitoring
disease occurrence and vaccine use. The established information systems, data
disease detection and control tools will be analysis capabilities, geographic dis-
introduced, distributed and harmonised. The tribution of virus strains, vector com-
consortium will also disseminate knowledge petence studies, entomological exper-
and train staff of relevant third countries. tise and surveillance methods applied,
The project partners think it is also impor- protocols for vaccine use) to serve as
tant to interlink different scientific disci- aframework for shared data sets.
plines which approach the problems from (d) Moreover, working groups will be estab-
different angles. lished to assess data focused on vector
control, vaccination and therapy. The
project will act as a platform to bring
Work plan together those participants who are
The work plan of Arbo-Zoonet foresees actively involved in molecular vaccine
anumber of interrelated tasks, with meas- development. Emphasis will be given
urable deliverables and milestones. The fol- to integrated vaccine strategies using
lowing are the specific aims of the work plan. vaccines based on pathogen and vec-
tor components and development of
(a) Identifying risk areas and undertak- appropriate delivery systems. In this
ing the necessary preparatory work context, studies on molecular charac-
for updated risk maps on RVFV, WNFV terisation the interaction between host,
and CCHFV introduction and/or spread vector and pathogen will be promoted,
throughout the EU territory. Efforts primarily through scientific exchange
will focus on understanding the ecol- visits. Therapeutic options will also be
ogy of host, vectors and disease res- examined. This will be done either by
ervoir. Moreover, this task will produce working on existing pharmaceuticals or
maps and estimate the numbers of by developing new ones.
vectors in order to prepare models for (e) The principal focus of the project is the
policymakers. transfer of knowledge and technology
(b) Create a pathogen database open to the between the members of the consor-
scientific community that will contain tium, which includes partners from rel-
information on where live samples of a evant countries outside the EU. In this
given pathogen are available. This data- context, links will be established to the
base will include other biological mater national and international organisa-
ial such as serum and genetic material tions (WHO, FAO, OIE, or the Interna-
from different geographical areas where tional Regional Organisation for Plant
the relevant diseases are endemic. and Animal Health (OIRSA)), institutions
(c) S urveillance networks will be estab- and laboratories located in the differ-
lished for the collection of global data ent areas in order to disseminate and
on the occurrence of RVFV, WNFV and transfer technologies needed to develop
CCHFV. An essential task is the reporting strategies for integrated control meas-
on the analysis of the RVFV, WNFV and ures in endemic regions such as diag-
CCHFV surveillance systems for the EU nostics, epidemiology and economic
and for affected areas in countries out- dimension of a number of endemic as
side the EU. These analyses will be used well as epizootic animal diseases.
to establish adequate georeferenced (f) Arbo-Zoonet will play a coordinating
data and to derive spatial conclusions. role a within the EUs animal health
The assessment will address significant strategy by bringing together interested
aspects of the surveillance and con- members of other EU consortia that
trol activities (monitoring approaches, share the focus on zoonoses caused by
diagnostic methods and capabilities, vector-borne arboviruses, such as the
Emerging Diseases in a changing Euro- Dissemination activities

pean eNvironment (EDEN) project, the The dissemination activities of the Arbo-
network of excellence for epizootic dis- Zoonet project comprised:
ease diagnosis and control (EPIZONE),
and the Environmental Vulnerability Publication and distribution of a bian-
Assessment (EVA) project. nual newsletter: Arbo-Zoonet news
Publication of an article in Eurosurveil-
lance, Vol. 14, Issue 12, 26March2009
Expected results Distribution of CDs containing presenta-
The coordinated research programme tions and protocols of the meetings and
comprising key laboratories in Europe and workshops conducted
neighbouring countries will address Maintenance of a specific website for
questions of joint interest, thus enabling this purpose (http://www.arbo-zoo.net)
the development of effective control meas-
ures that will improve the EUs response to
disease outbreaks. Keywords
Crimean-Congo hemorrhagic fever, Rift
Valley fever, West Nile fever, arbovirus,
Potential applications hemorrhagic fever, encephalitis, mosquito,
The Arbo-Zoonet consortium hopes this tick
network will be continued beyond the time
frame of the project, and will be extended Coordinator
to address other public and animal health Dr Michle Bouloy
problems. Institut Pasteur
2528 rue du Dr Roux
75724 Paris
Activities FRANCE
A number of activities such as scientific mbouloy@pasteur.fr
meetings and technical workshops both on
national and regional levels were organised.
In summary, a total of six scientific meet-
ings and technical workshops for epidemi-
ology and diagnostic tools were held in the
following countries: Algeria, France, Ger-
many, South Africa and Turkey.
In the context of collaboration, the Arbo-

Zoonet project established the following
links:
FAO (North Africa)

EPIZONE
International Consortium on Ticks and
Tick-Borne Diseases (ICTTD-3)
ConFluTech
ASEMDialog
Society for Tropical Veterinary Medicine
European Meeting on Viral Zoonoses,
Saint Raphal, France, September 2009
Partners
Research Center Borstel GERMANY J. Ahmed jahmed@fz-borstel.de
Borstel
Veterinary Laboratories UNITED T. Fooks t.fooks@vla.defra.gsi.gov.uk
Agency KINGDOM
Weybridge
Centre de coopration FRANCE V. Chevalier veronique.chevalier@cirad.fr
internationale
en recherche agronomique
pour le dveloppement
Montpellier
University of Bonn GERMANY C. Drosten drosten@bni-hamburg.de
Bonn
Swedish Institute for SWEDEN A. Mirazimi ali.mirazimi@smi.ki.se
Infectious Disease Control
Solna Municipality
Istituto Zooprofilattico ITALY P. Calistri p.calistri@izs.it
Sperimentale dellAbruzzo
e del Molise G. Caporale
Teramo
University Ljubljana SLOVENIA T. Avsic-Zupanc tatjana.avsic@MF.UNI-LJ.SI
Institute of Microbiology
and Immunology
Ljubljana
National Health Laboratory SOUTH AFRICA J. Paweska januszp@nicd.ac.za
Services
Johannesburg
Health Protection Agency UNITED J. Medlock jolyon.medlock@hpa.org.uk
Porton Down KINGDOM
Lanzhou Veterinary CHINA H. Yin yinhong@public.lz.gs.cn
Research Institute
Lanzhou
Pasteur Institute of Iran IRAN C. Sadegh chinikar@pasteur.ac.ir
Tehran
Robert Koch Institute GERMANY M. Niedrig NiedrigM@rki.de
Berlin
Veterinary and Agrochemical BELGIUM K. De Clercq krdec@var.fgov.be
Research Centre
Brussels
Stichting Dienst NETHERLANDS R. Moorman Rob.Moormann@wur.nl
Landbouwkundig Onderzoek
Wageningen
Institut Pasteur de Dakar SENEGAL A. Sall asall@pasteur.sn
Dakar
Universidade Nova de Lisboa PORTUGAL P. Almeida palmeida@ihmt.unl.pt
Lisbon
Instituto Nacional de SPAIN A. Brun brun@inia.es
Investigacin y Tecnologa
Agraria y Alimentaria
Madrid

Aristotle University GREECE A. Papa-Konidari annap@med.auth.gr
of Thessaloniki
Thessaloniki
Istituto Superiore di Sanit ITALY F.M. Ruggeri franco.ruggeri@iss.it
Rome
University of Zaragoza SPAIN A. Estrada-Pea aestrada@unizar.es
Zaragoza
University Court of the UNITED R.M. Elliott rme1@st-andrews.ac.uk
University of St Andrews KINGDOM
St Andrews
Marmara University TURKEY O. Ergonul onderergonul@yahoo.com
Istanbul
Kafkas University TURKEY Z. Vatansever zativet@gmail.com
Kars
[WildTech]
Novel Technologies for

Surveillance of Emerging
and Re-emerging
Infections of Wildlife
Summary The project combines
Acronym:
For many reasons, the health of wildlife is of technological development to enable
WildTech
major concern throughout the world. Apart high throughput nucleic acid- and
from important influences on the health of peptide-based array screening of Project number:
222633
many wildlife species, infectious diseases samples from a wide variety of wild
of wildlife have significant impacts on pub- animals; EC contribution:
lic health and health of livestock. Effective surveillance of terrestrial, aerial and EUR 5 996 822
disease surveillance is essential in order to marine wild animal species within Duration:
inform control strategies and this depends Europe and from countries which 48 months
critically on the development and applica- actasportals of disease entry into Start date:
tion of methods of disease diagnosis which theEU; 1 July 2009
are both accurate and rapid. The WildTech epidemiological analysis and risk assess- Instrument:
project has been established specifically ment using data generated duringthe Collaborative project
to address these problems and to set up a project and from other sources;
technology centre that may be exploited in development and proposal of a model
Europe and elsewhere as a basis for high framework for disease surveillance
throughput disease diagnosis in wildlife. within Europe.
The project will place the EU at the centre of wildlife origin. It is therefore clear that
of wildlife disease surveillance and ena- the surveillance of disease in wildlife not
bles the translation of high throughput only impacts on communities that rely on
array-based technologies to human and healthy domestic animals but is also an
veterinary medicine. essential tool for the protection of human
health. Despite this alarming situation, sur-
veillance for infectious diseases in wildlife
is far from satisfactory. Until now, there has
been no coordinated effort to monitor the
spread of infection within and between dif-
ferent countries in the EU. Surveillance of
wildlife infectious disease has been largely
passive in structure rather than a proactive
attempt to predict and manage future dis-
ease threats across Europe.
Aim
The key objectives of WildTech are as
follows.
The application of microarray technology

Problem for the detection of known infectious
WildTech addresses the problem of the agents in wildlife populations.
increasing prevalence of new and emerg- The application of microarray technol-
ing diseases arising from wildlife which ogy to the detection and identification of
has clear implications for disease spread to novel and unknown infectious agents in
domestic animals and humans both across wildlife populations.
Europe and globally. The reasons for this The application of microarray technol-
alarming trend are multifactorial and have ogy to the development of high through-
been well documented in the literature. In put serological screening of wildlife
brief, the continued increase in the human populations for infectious disease.
population results in habitat fragmentation The utilisation of these technologies
caused by factors such as deforestation and toassess the spread of selected diseases
increasing levels of pollutants. These issues (proof of concept) using historical samples
inevitably impact on host-pathogen rela- and those collected during the project. We
tionships and the spread of pathogens into will monitor and model patterns of wild-
geographical areas previously unaffected. life disease spread and the risks associ-
Alterations in land use and livestock rearing ated with it. Ultimately, this epidemiology
practices and the rapid global movement framework will be used to reduce the risk
of humans, animals and other organisms of further potential epidemics bypro-
are also key factors. Finally, the evolution ducing ageneric action plan in case of
of viral pathogens adds another factor into emerging epizootics among wildlife.
this alarming and complex equation. The development of a state-of-the-art
wildlife disease data management sys-
WildTech is focused on wildlife as a reser- tem with mapping capability for use in
voir of disease. It is reported that 61 % of Europe and beyond.
known pathogens infect multiple animal The establishment of a framework for
species and 75 % of all diseases which pan-European surveillance of wildlife
have emerged in the last two decades are diseases.
Expected results under the aegis of the OIE will make a

Effective and validated high throughput substantial contribution to preventing
microarray technology, both generic and major outbreaks of infectious disease
adapted to a commercial platform, for in Europe.
the detection of nucleic acid of a focused
list of up to 20 infectious agents (viruses, 2. The benefits to the European economy
bacteria and parasites) from wild animal also include short-term direct eco-
samples. We will, in addition, develop nomic benefits from the exploitation
generic arrays for 200 infectious agents and application of the technologies to
which will be incompletely validated. be developed during this project. The
Effective and validated high through- exploitation of the high throughput
put serological array technology, both microarray technology will enable a
generic and partially adapted to com- European company to be a major focus
mercial platform for detection of spe- for high throughput disease-screening
cific antibodies in serum/blood against technology. This microarray technology
approximately 20infectious agents from will expand the current income stream
selected wild animal hosts, in addition to from within and outside Europe.
incompletely validated tests for further
infectious agents. 3. The exploitation of the serology array
Information on the spatial and temporal will enable expansion of the exist-
distribution of a focused list of up to 20 ing market in serological detection of
infectious agents in wild animal species food-borne zoonoses and will facili-
in selected European countries/regions tate the development of new markets
and countries outside Europe that rep- in surveillance of wildlife and other
resent potential sources of introduction animal and human diseases. The size
into Europe. of this market is impossible to predict
Information on the risk to human and and will depend on the extent to which
domestic animal health from the pres- large-scale surveillance of diseases
ence and evolution of infectious agents in wildlife, man and domestic and
in selected wild animal populations. companion animals becomes a real-
Established management systems for ity within Europe and beyond. Initial
wildlife disease information, which are markets are likely to be in Europe and
accessible to national and international North America.
animal and human health organisa-
tions, the international wildlife disease
community and policymakers. Social impact
Proposal for surveillance system for wild- 1. The enormous financial losses from
life diseases in Europe, which will con- emerging animal diseases hide the
tribute to protecting European wildlife, intense personal impact that such dis-
domestic animal and human health. eases have on rural and other com-
munities, both through the destruction
Potential applications of animals and livelihoods and, in the
Benefits to the European economy include case of zoonoses, through the potential
the following. for large-scale human morbidity and
mortality with the associated economic
1. The technology proposed for this pro- and personal damage inflicted on the
ject together with the strengthened continent. The proposed combination of
European wildlife community net new generation technologies together
workworking in collaboration with with an improved framework for moni-
international reference laboratories toring and surveillance are central to
ensuring that major outbreaks do not through wildlife are likely sources of
occur with the potentially devastat- zoonotic infection. By improving our
ing effects on the European and world detection of these pathogens, this
economy and to developing a mecha- would enable a rapid and effective
nism that can effectively identify and response to an emerging infection,
respond to the threat presented by new which would minimise the impact on the
pathogens. human population.
2. The European effects will be mirrored

by reduced mortality and morbidity Project website
and associated improved welfare in http://www.wildtechproject.com
domestic animals beyond Europe. In
poorer third world countries, outbreaks Keywords
of infectious disease in domestic ani- wildlife disease surveillance, high through-
mals can have far-reaching conse- put array technologies, epidemiology, data
quences for the well-being of entire management systems
human communities. Thus, improved
surveillance and subsequent inter- Coordinator
vention strategies will have dramatic University Park Nottingham
effects on the quality of human life in NG7 2RD
more deprived areas within and out- UNITED KINGDOM
side Europe. duncan.hannant@nottingham.ac.uk
3. The project results will also have indi-

rect impact on human health as dis-
eases coming either directly or indirectly
Partners
ORGANISATION ADDRESS CONTACT E-MAIL
The University
of Nottingham
School of Vet- School of Veterinary Medicine Duncan Hannant duncan.hannant@not-
erinary Medicine and Science tingham.ac.uk
and Science University of Nottingham
(Coordinator) Sutton Bonington Campus
Loughborough,
LE12 5RD
UNITED KINGDOM
School of Intelligent Modelling Uwe Aickelin uwe.aickelin@notting-
Computer Science & Analysis Research Group ham.ac.uk
(IMA)
School of Computer Science
The University of Nottingham
Jubilee Campus
Wollaton Road
Nottingham,
NG8 1BB
UNITED KINGDOM

Institute for Animal Ash Road, Peter Mertens peter.mertens@bbsrc.
Health, Pirbright Pirbright, Surrey ac.uk
GU24 0NF
UNITED KINGDOM
Friedrich-Loeffler- Naumburger Str. 96a Konrad Sachse konrad.sachse@fli.
Institut, Jena 07743 Jena bund.de
GERMANY
Central Veterinary P.O. Box 65 Alex Bossers Alex.Bossers@wur.nl
Institute of Wage- 8200 AB Lelystad
ningen University NETHERLANDS
and Research
Centre
Veterinary Labora Woodham Lane Liljana Petrovska l.petrovska@vla.defra.
tories Agency, New Haw, Addlestone, Surrey gsi.gov.uk
Weybridge KT15 3NB
UNITED KINGDOM
Alere Technologies Loebstedter Str. 103105 Ralf Ehricht ralf@clondiag.com
GmbH 07749 Jena
GERMANY
Center for Papanastasiou str. 51 Charalambos billinis@vet.uth.gr
Research and Tech- 41222 Larissa Billinis
nology Thessaly GREECE
National SE-751 89 Uppsala Dolores dolores.gavier-widen@
Veterinary SWEDEN Gavier-Widn sva.se
Institute
VetAgro Sup: 1 Avenue Bourgelat Marc Artois m.artois@vetagro-
Institute for Higher 69280 Marcy LEtoile sup.fr
Education and FRANCE
Research in Food,
Animal Health,
Agronomic and
Environmental
Sciences
Scottish Kings Buildings, West Mains Mike Hutchings mike.hutchings@sac.
Agricultural Road ac.uk
College Edinburgh
EH9 3JG
UNITED KINGDOM
University of 52 Campus Drive Ted Leighton ted.leighton@usask.ca
Saskatchewan Saskatoon
SK S7N 5B4
CANADA
Twycross Zoo Burton Road Suzanne suzanne.i.boardman@
East Midland Zoo- Atherstone Boardman twycrosszoo.org
logical Society Warwickshire,
CV9 3PX
UNITED KINGDOM
[ICONZ]
Integrated control of neglected

zoonoses: improving human
health and animal production
through scientific innovation
and public engagement
Acronym: Summary trypa nosomiasis. Through a series of
ICONZ ICONZ is a five-year, 21-partner EU FP7 complementary actions, ICONZ aims to
Project number: project undertaking studies to evalu- improve human health and animal pro-
221948 ate the epidemiology, socioeconomic and duction to alleviate poverty and contrib-
EC contribution: policy implications of eight neglected ute to the Millennium Development Goals
EUR 6 million zoonotic diseases across seven African (MDGs). New technologies and research
countries. outcomes are being used to quantify
Duration:
5 years burdens of neglected zoonoses on com-
munities in Africa through a series of
Start date:
April 2009
Problem case studies in the International Part-
Neglected zoonoses are major causes of ner Cooperation Countries (ICPCs); Mali,
Instrument: ill health in many developing countries Morocco, Mozambique, Nigeria, Tanzania,
Collaborative project
worldwide. Both production and compan- Uganda and Zambia. It is expected that
ion animals can act as significant reser- through these field-based case stud-
voirs for transmission of zoonotic disease ies, improved control and awareness of
to man. This societal burden of disease is neglected zoonotic disease at both com-
often compounded by the effect of zoonotic munity and national/regional policy levels
disease on livestock productivity, hence will occur through the development and
inflicting a further burden on affected com- utilisation of novel intervention strategies.
munities who depend on their livestock as It is expected that intervention strate-
asource of food and income. gies will be communicated through a well
researched dissemination strategy to both
communities and policymakers in affected
Aim regions.
ICONZ is a collaborative project involv-
ing 21 European and African partners,
with the integrated control packages of Expected results
eight key neglected zoonoses, namely It is widely accepted that current estimates
anthrax, bovine tuberculosis, brucellosis, of the prevalence and burden of neglected
cystic echinococcus, porcine cysticerco- zoonotic diseases are inadequate. ICONZ
sis, leishmaniasis, rabies and zoonotic anticipates and predicts a major impact
on the understanding, implementation and for improved inputs. The development of

efficacy of control strategies for neglected cost-effective diagnostic and control tools
zoonoses in developing countries, through which are field-friendly and easily avail-
(i) increased understanding of their burden able cannot be underestimated: effective
and significance and (ii) through the test- tools employed within simple strategies
ing and refinement of practical, cost-effec- may preclude the expense of emergent
tive strategies for control. To support this, tools, vaccines and improved veterinary
ICONZ is currently in the process of under- medicines.
taking surveys across seven countries in
Africa, in order to increase understand-
ing of the societal burden of neglected Project website
zoonoses, and help develop informed http://www.iconzafrica.org/
control strategies, taking into considera-
tion the socioeconomic status of affected Keywords
communities, and the wider national and neglected zoonoses, one health, livestock
regional policy agendas that govern these productivity, human health
communities.
Logo
The ongoing overview of current worldwide
research activities aims to reduce duplica-
tion of effort, leading to more effective use
of limited resources and funds, whilst at the Coordinator
same time identifying major gaps within Prof. Sue Welburn
international research agendas. ICONZ is Univesity of Edinburgh
currently also in the process of catalogu- Chancellors Building
ing current tools for surveillance, diagno- 49 Little France Crescent
sis and treatment of neglected zoonoses, Edinburgh EH16 4SB
which will help identify areas where such UNITED KINGDOM
tools are inadequate or unavailable, and sue.welburn@ed.ac.uk
subsequently lead to recommendations
Partners
ORGANISATION ADDRESS CONTACT EMAIL
Institute of Tropical Nationalestraat 155 Pierre Dorny pdorny@itg.be
Medicine 2000 Antwerpern
BELGIUM
University of 10 Noerregade Maria Vang Johansen mvj@life.ku.dk
Copenhagen 1017 Koebenhavn K
DENMARK
Agence nationale de 2731 Avenue du Franck Boue Franck.BOUE@anses.fr
scurit sanitaire Gnral Leclerc
de lalimentation, de 94701 Maisons-Alfort
lenvironment et du FRANCE
travail

Universit Claude 43 boulevard du 11 Anne-Francoise petavy@univ-lyon1.fr
Bernard Lyon 1 Novembre 1918, Petavy
69622 Villeurbanne
FRANCE
Friedrich-Loeffler- Sudufer 10 Manfred Tanner manfred.tanner@fli.
Institut, Federal 17493 Greifswald- bund.de
Research Institute for Insel
Animal Health Riems
GERMANY
University of Minho Largo Do Paco Margarida mcorreianeves@
4704-553 Braga Correida-Neves ecsaude.uminho.pt
PORTUGAL
University of Navarra Carrer Logrono S/N Ignacio Moriyon imoriyon@unav.es
31080 Pamplona
SPAIN
Karolinska Institutet Nobels vag 5 Gunilla Kallenius Gunilla.kallenius@ki.se
SE-171 77 Stockholm
SWEDEN
Swiss Tropical and Socinstrasse 57 Jakob Zinsstag jakob.zinsstag@
Public Health Institute Case Postale unibas.ch
4002 Ble
SWITZERLAND
University of Liverpool The Foundation Jim Scudamore j.scudamore@liv.ac.uk
Building
Brownlow Hill
Liverpool L69 7ZX
UNITED KINGDOM
Laboratoire Central Km8 Route de Saidou Tembely stembely@yahoo.com
Vtrinaire Koulikoro
Bamako 2295
MALI
Institut Agronomique Avenue Allal el Fassi Allal Dakkak a.dakkak@iav.ac.ma
et Vtrinaire Madinat Al,
Hassan II 10101 Rabat
MOROCCO
Eduardo Mondlane 257 Raitoria da Sonia Afonso santanafonso@yahoo.
University Universidade co.uk
Praca 25 de Junho
Maputo
MOZAMBIQUE
National Veterinary Vwang Road Reuben Ocholi Rbocholi@yahoo.com
Research Institute, 930010 Vom
Vom NIGERIA
Makerere University PO Box 7062 Charles Waiswa cwaiswa@vetmed.
Kampala, UGANDA mak.ac.ug
SACEMA, Stellenbosch Private bag x1 John Hargrove jhargrove@sun.ac.za
University Matieland
7602 Stellenbosch
SOUTH AFRICA

Sokoine University Sokoine University Rudovick Kazwala Kazwala@suanet.ac.tz
of Agriculture of Agriculture
Morogoro
TANZANIA
University of Zambia Great East Road Chummy Sikasunge chumsika@yahoo.co.uk
Campus
10101 Lusaka
ZAMBIA
AVIA-GIS 33 Risschotlei Guy Hendrickx ghendrickx@avia-gis.
2980 Zoersel be
BELGIUM
International Old Naivasha Road John McDermott j.mcdermott@cgiar.org
Livestock Research 00100 Nairobi
Institute KENYA
41
CHAPTER 2.
Epidemic diseases of
livestock
European policies for animal

health sustained by EC-
funded research projects
Research to support animal health is crit currently underway requires an interdis-
ical for the development of evidence-based ciplinary and collaborative approach. It is
policies and the introduction of legislative no longer the preserve of one discipline
measures to govern disease surveillance, and a wide range of multi- and interdis-
prevention, control and eradication. These ciplinary research activities are required
policies must be based on the best scientific to solve complex problems. In addition to
evidence, some of which is provided from researchers and scientists, it is increasingly
the portfolio of projects funded through important to imply the whole spectrum of
the EU seventh framework programme for expertise involved in animal health includ-
research (200713). ing producers, advisors, veterinarians and
scientists. This is encouraged by a number
Animal diseases of public importance of EC-funded projects which will generate
remain a priority area but research funding durable interactions among partners with
is not unlimited. The DISCONTOOLS project complementary expertise and skills.
provides a mechanism to prioritise diseases
and to identify the gaps in current know Research is needed to ensure that the facil
ledge and available control tools such as ities and expertise necessary for the investi-
vaccine, diagnostic tests and pharmaceut gation and identification of new and emerg-
icals. The coordination of research funding ing diseases is available. WILDTECH develops
between the EC projects and those funded more effective surveillance methods for
by the Member States is essential to obtain emerging diseases from wildlife. Another
maximum benefits, from the available forward-looking project, ARBO-ZOONET,
funding. The EMIDA and STAR-IDAZ projects builds capacity so that Europe would have the
will both make important contributions by expertise and facilities to deal with the poten-
coordinating European research and devel- tial risks of disease outbreaks as a result of
oping EU-wide and global networks of fund- emerging viral vector-borne diseases.
ing organisations.
It is also quite important to support
Some overarching projects seek to develop research in neighbouring countries and
networks to maximise the use of expertise, developing countries especially for exotic
increase excellence through collaboration diseases of high prevalence which have the
and ensure that Europe has the capacity potential to enter the EU. There are many
to deal with emerging problems. Networks benefits from the range of EC-funded pro-
and collaboration are vital as it is no longer jects which involve research institutes from
feasible for one group alone to undertake countries outside the EU.
complex and expensive research.
New or improved tools for the prevention
Even for targeted diseases such as foot- and control of animal diseases are an essen-
and-mouth (FMD), much of the research tial component of EC research programmes.
C H A P T E R 2 . E P I D E M I C D I S E A S E S O F L I V E S T O C K 43
Diagnostic tests with high sensitivity, speci- Member States and in non-EU countries
ficity and reproducibility are requested for engaged in trading of livestock and products
use on the front line, local laboratories and of animal origin. Evidence from EU research
in abattoirs. New techn ologies enable the is often used by international bodies such as
development of pen-side tests for a range of the World Organisation for Animal Health
diseases with one sample along with remote (OIE) to develop international standards for
transfer of data. Projects such as CSFVAC- disease control, animal welfare and trade.
CINE & WILD BOAR, CSF-GoDIVA or FMD-
Disconvac have resulted in the development Much of the research into animal diseases
of diagnostic tests which could also dis- is carried out in support of policies by pro-
criminate between vaccinated and infected viding the scientific evidence and technolo-
animals. This is critical information for the gies to those responsible for policy devel-
development of new control policies. opment. This is a particularly important
pathway in animal health and welfare and
The development of new or improved vac- has been used very effectively over the
cines for a wide range of epidemic and past decade. The EC-funded research pro-
endemic diseases was the topic of a num- jects have made major contributions to
ber of research projects. In many cases, the European policies for animal health with a
research aimed to understand the immune comprehensive portfolio in relation to policy
response, to develop improved vaccines needs. The programme has also been char-
which would provide protection against a acterised by the anticipation of research
number of serotypes and to use third gen- needs in advance of these becoming clear
eration technology to develop marker vacin policy circles. This is a major achievement
cines. Research projects were funded to and demonstrates the benefit of long-term
develop vaccines for bluetongue (BT-VAC strategic portfolio management by an indi-
ORBIVA), classical swine fever (CSFVAC- vidual Dr Isabel Minguez-Tudela who had
CINE & WILD BOAR and CSF-GoDIVA), FMD the foresight to identify future research
(FMD Disconvac) and African horse sickness needs and to ensure continuity of research.
(ORBIVAC). In a number of these projects,
the potential for control measures using Jim Scudamore
vaccines was also studied in order to reduce Professor of Livestock
the need to slaughter animals during an and Veterinary Public Health
outbreak of an epidemic disease. School of Veterinary Science,
University of Liverpool
Projects such as those for African swine
fever, avian influenza, bluetongue, classi- Alberto Laddomada
cal swine fever and FMD have significant Deputy Head of Unit Animal Health
impacts. The scientific outputs often result and Standing Committees
in improved measures being adopted in EU DG Sanco, European Commission
2.1. Foot-and-mouth disease

[FMD_ImproCon]
Improvement of foot-and-
mouth disease control by
ethically acceptable methods
based on scientifically validated
assays and new knowledge
on FMD vaccines, including
the impact of vaccination
Acronym: Summary to discriminate unequivocally between
FMD_ImproCon There is a strong desire to reduce reliance infected and vaccinated animals, in order
on large-scale culling of animals to con- to allow the implementation of the new
Project number:
503603 trol future outbreaks of FMD in EU Mem- policy in the immediate term. Validation
ber States. As an alternative, it is proposed of existing and new NSP tests as con-
EC contribution:
EUR 2 399 907
to use emergency vaccination and then to firmatory tests will be a major output
screen for residual infection using tests of this project. The experimental design
Duration: for antibodies to the non-structural pro- will also provide expected outputs in the
60 months
teins of FMD virus. It is intended to amend field of the impact of vaccination on the
Start date: the policy on FMD control to enable such carrier state and on virus dissemination,
1 January 2004
an approach to be used in the very near the onset of vaccinal protection, vaccine
Instrument: future. In reality, this means that current potency in relation to emergency use,
Specific Targeted contingencies must be based on the use vaccine strain selection and new marker
Research
of existing vaccines. Therefore, this pro- vaccines. This project focuses on marker
or Innovation Project
(STREP) ject seeks to address the specific gaps in vaccines to induce durable protection
our knowledge and technological ability against FMD. Conventional and marker
with respect to the implementation of a vaccines will be targeted to dendritic cells
vaccinate-to-live policy. The availabil- with particular attention to promote den-
ity of adequate discriminatory diagnos- dritic cell mucosal homing (from parental
tic tests is the keystone of the new EU immunisation), because mucosal immunity
FMD control policy. The project is focused can prevent FMD virus establishing local
on the validation of NSP-based tests infection and the carrier status.
Problem Knowledge (advantages and limitations)

As there is a strong desire to reduce reli- on their sensitivity and specificity, as well
ance on large-scale culling of animals as on application of such tests in outbreak
to control future outbreaks of foot-and- and post-outbreak situations, is essential in
mouth disease (FMD) in EU Member States, light of the requirements laid down in the
the European Commission (EC) amended new Council Directive on Community meas-
its policy and has changed its directive on ures for the control of foot-and-mouth dis-
Community measures for the control of ease (Council Directive 2003/85/EC) and
foot-and-mouth disease (Council Directive their application to substantiate freedom
2003/85/EC), making the use of emergency from FMD has been discussed.
vaccination easier when combined with
screening for residual infection using tests A tool for validating NSP tests without Gold
for antibodies to non-structural proteins Standard was provided by: (i) the develop-
(NSPs). In reality, this means that current ment of at least four tests based on differ-
contingencies must be based on the use ent detection methods for the virus antigen
of existing vaccines. Therefore, this project (for instance an antigen-detection ELISA,
addressed the specific gaps in our knowl- virus isolation, PCR and a RCA-ELISA (see
edge and technological ability with respect below)); or by (ii) the use of state-of-the-
to the implementation of a vaccinate-to- art statistical methods such as Latent Class
live policy. analysis or Bayesian analysis.
WP2: Development and validation

Aim of confirmatory and new NSP tests
The aim of the project was to support the The work performed in WP2 was aimed at
EC policy to reduce reliance on large-scale developing and validating new NSP assays
culling of animals for controlling future as confirmatory tests or as alternative pri-
FMD outbreaks in EU Member States and mary assays, in order to improve on the
to increase the possibility for implementing diagnostic performance of the NSP ELISAs
the vaccinate-to-live policy through: validated under WP1 and to create kit pro-
duction opportunities of NSP assays with
v a l id at io n a n d d e vel o p m e n t of a view to facilitate the distribution of the
(confirmatory) NSP tests; product to all interested countries within
improved FMD virus detection; the European Union and the rest of the
better knowledge on the impact of vac- world. Some of the developed prototype
cination on FMD virus dissemination and assays have already been taken up by kit
the carrier state; manufacturers and others might do this in
improved vaccine strain selection; the future.
the development and evaluation of FMD
marker vaccines; WP3: Improved FMD virus detection
better knowledge on mucosal immune Improving on existing FMD detection meth-
responses and dendritic cell targeting of ods in WP3 has sometimes proven diffi-
the FMD vaccines. cult, but by better understanding the basic
principles of FMD growth on cell culture,
Results progress has been made.
WP1: Validation of existing NSP
tests A real-time RT-PCR system consisting of
WP1 of the project has contributed sig- two independent FMDV PCR protocols (plus
nificantly to the availability of validated one backup protocol) and two independent
assays able to discriminate unequivocally SVD PCR protocols were established and
between vaccinated and infected animals. validated.
Furthermore, a new goat cell line has been interferon-gamma responses and antigen
identified to enhance FMD virus detection payload).
that also enables a quicker diagnosis.
Mouse monoclonal Antibody (MAb) anti-
WP4: Impact of vaccination on virus genic profiling data proved to be a very
dissemination and the carrier state effective tool for monitoring how effec-
WP4 provided new insights in virus trans- tively a vaccine strain protects against
mission and the impact of vaccination, field viruses. However, mapping data com-
which will prove crucial when faced with bined with full capsid sequences of field
new FMD outbreaks. The value of vaccin strains would provide more information
ation in reducing virus transmission has which helps to resolve the crucial epitopes
been demonstrated in different FMD sus- and develop vaccine strategies focus-
ceptible species (cattle, sheep and pigs) ing on these epitopes. Moreover, further
with different FMDV strains and serotypes. research is required to better understand
However, the results of WP4 also clearly the contribution of different viral epitopes
show that vaccination will not be able to to cross-protection and thereby to improve
prevent infection when it occurs within a predictive methods of vaccine efficacy.
week following the administration of the
vaccine. Nevertheless, these experiments It is well known that neutralising antibody
were successful in measuring the efficacy titres are important in protecting against
of vaccines, the dynamics of NSP serocon- FMDV infection. However, it has often been
version and its relationship to virus replica- shown that the humoral antibody titre is not
tion, persistence and clinical signs. Math- always fully predictive of vaccine-induced
ematical analysis of the experimental data protection against FMD. Therefore, a cor-
has been used to predict carrier numbers in relation between cell-mediated immune
cattle herds and sheep flocks after emer- responses, humoral immune responses and
gency vaccination, as an aid to determining post-vaccination protection against FMDV
the feasibility of using NSP serosurveillance infection was investigated. It is concluded
to substantiate FMD freedom. that T cell stimulation assays such as the
whole blood IFN- assay along with VNT
WP5: Improved vaccine strain are potential candidates for vaccine evalu-
selection ation and could reduce the need for in vivo
The data provided on vaccine strain selec- challenge in the future.
tion in WP5 will help decision-makers in
their difficult choice of vaccine use and in WP6: Development of a marker
identifying relevant strains for inclusion vaccine
in FMDV antigen banks. The advantage Progress has been made in WP6 to enhance
of using high potency vaccines when no mucosal immunity. Moreover, a new gen-
matching vaccines are readily available eration vaccine based on a serotype A
in antigen reserves has been shown. High chimera vaccine in which the GH-loop
potency vaccines enhance the probabil- region was replaced with that of another
ity of achieving adequate levels of cross- serotype proved fully protective in cattle
protection even when faced with low r-val- against challenge from the unsubstituted
ues and sequence homology. Furthermore, parental serotype A virus. Furthermore, the
the limits of the existing in vivo FMD vac- presence of a heterologous GH-loop could
cine potency tests have been demonstrated be exploited to discriminate serologically
(high level of between test variability) which between vaccinated and vaccinated-and-
point towards more international accept- challenged cattle. Subsequently, a sponta-
ance of alternative in vitro approaches neous loop-deleted mutant vaccine virus
avoiding live viral challenge (e.g. serology, was discovered and shown to elicit an
antibody response that was similarly cross- continents. Workshops have been organ-
reactive compared to antibodies elicited by ised in collaboration with the Directorate-
a loop-undeleted vaccine virus. Follow-up General for Research and Innovation and
funding has been granted to further pursue the Directorate-General for Health and
the marker vaccine potential of this virus. Consumers for the EU reference labo-
ratories, in collaboration with other FP6
Dendritic cells (DC), essential for induc- projects (EPIZONE and CA FMD/CSF), FAO-
ing and regulating immune defences and EUFMD, OIE and TAIEX for candidate Mem-
responses (systemic and mucosal), repre- ber States. Results will further be discussed
sent the critical target for vaccines against with the Directorate-General for Research
pathogens such as FMDV. The interaction of and Innovation and the Directorate-General
FMDV vaccine antigen with DC was stud- for Health and Consumers. If results are
ied and showed that following internalisa- deemed appropriate to change the control
tion of FMDV antigen, these DC were effi- policy, they could be presented to the EC
cient antigen presenting cells, observed in Scientific Committee. Through close col-
terms of their ability to stimulate specific laboration between some of the partners
lymphocyte proliferation and virus-specific and the EU, FAO-EUFMD and the OIE, possi-
antibody production. These results are ble changes can be communicated to these
advantageous for conventional FMD vac- organisations.
cines, which will be composed cell-culture
adapted HS-binding variants of FMDV. WP1 has contributed to the availability of
Furthermore, immuno-modulatory factors validated assays to be applied in the sub-
targeting DC for promotion of antibody and stantiation of freedom from FMD when vac-
mucosal immune responses, like the vita- cination is performed and clear advice for
min A derivative all-trans-retinoic acid and the control policymakers on the application
the E. coli-derived heat-labile enterotoxin of NSP tests has been expressed.
(LT), were identified. Moreover, the use of
TLR ligands as additional immunostimu- Some of the prototype assays developed
lating molecules to promote systemic and within WP2 have already been taken up by
mucosal immune responses was charac- kit manufacturers and others might be in
terised in vitro and in vivo. Nevertheless, the future.
caution is required when translating find-
ings from mouse models to a natural host A new goat cell line has been identified
of FMD. within WP3, to enhance FMD virus detection
that also enables a quicker diagnosis.
Applications The animal experiments performed in WP4

New diagnostic experience and new and WP5 provided indispensable serum
knowledge about the epidemiology and sample collections from vaccinated and/or
virus properties were transferred in dif- infected animals that enabled validation
ferent ways. Results open to the public and development of existing and new DIVA
were placed on the website (http://www. (differentiation of infected from vaccinated
fmdimprocon.org) and presented not only animals) tests. These sera can readily be
at scientific meetings of the project group, shared with other institutes and organisa-
but also at the annual meeting of the EU tions, especially since reference serum pan-
reference laboratories for vesicular dis- els have been developed for the purpose of
eases and at the open sessions of the FAO- assessing FMD DIVA tests.
EUFMD RG that brings together, every two
years, 32 European countries and reference The data provided on vaccine strain selec-
laboratory representatives from all other tion in WP5 will help decision-makers in
their difficult choice of vaccine use and in virus transmission among dairy cows
identifying relevant strains for inclusion Vaccine, 25(2): 32735.
in FMDV antigen banks. Furthermore, the
limits of the existing in vivo FMD vaccine Parida, S., Fleming, L., Oh, Y., Mahapatra,
potency tests have been demonstrated M., Hamblin, P., Gloster, J., Doel, C., Gub-
(high level of between test variability) which bins, S., Paton, D.J. (2007), Reduction of
point towards more international accept- foot-and-mouth disease (FMD) virus load
ance of alternative in vitro approaches in nasal excretions, saliva and exhaled air
avoiding live viral challenge (e.g. serology, of vaccinated pigs following direct contact
interferon-gamma responses and antigen challenge, Vaccine 25(45): 780617
payload).
Progress has been made in WP6 to enhance Keywords

mucosal immunity. Moreover, a new gen- foot-and-mouth disease, non-struc-
eration vaccine based on a serotype A chi- tural proteins, vaccine, cross-protection,
mera vaccine in which the GH-loop region validation, mass culling
was replaced with that of another serotype
proved fully protective in cattle against Project website
challenge from the unsubstituted parental http://www.fmdimprocon.org
serotype A virus.
Coordinator
Dr Kris De Clercq
References/publications Centrum voor Onderzoek in Dierge-
Brehm, K.E., Kumar, N., Thulke, H.-H., Haas, neeskunde en Agrochemie
B. (2008), High potency vaccines induce CODA
protection against heterologous challenge Groeselenberg 99
with foot-and-mouth disease, Vaccine, 26: 1180 Ukkel
16811687. BELGIUM
krdec@var.fgov.be
Brocchi, E., Bergmann, I., Dekker, A., Paton,
D.J., Sammin, D.J., Greiner, M., Grazioli, S.,
De Simone, F., Yadin, H., Haas, B., Bulut, N.,
Malirat, V., Neitzert, E., Goris, N., Parida, S.,
Sorensen, K., De Clercq, K. (2006), Com
parative evaluation of six ELISAs for the
detection of antibodies to the non-struc-
tural proteins of foot-and-mouth disease
virus, Vaccine, 24(4748):69666979
Goris, N., Merkelbach-Peters, P., Diev, V.I.,

Verloo, D., Zakharov, V.M., Kraft, H.-P., De
Clercq, K. (2007), European Pharmaco-
poeia foot-and-mouth disease vaccine
potency testing in cattle: Between test vari-
ability and its consequences, Vaccine, 25:
33733379.
Orsel, K., de Jong, M.C.M., Bouma, A., Stege-

man, J.A., Dekker, A. (2007), The effect of
vaccination on foot-and-mouth disease
Partners
Institute for Ash Road Dr David Paton david.paton@bbsrc.
Animal Health (IAH) Pirbright, Surrey, ac.uk
GU24 ONF
UNITED KINGDOM
Central Veterinary P.O. Box 65 Dr Aldo Dekker aldo.dekker@wur.nl
Institute of 8200 AB Lelystad
Wageningen UR (CVI) NETHERLANDS
Danmarks Lindholm Dr Laurids Siig LSI@food.dtu.dk
Fodevareforskning 4471 Kalvehave Christensen
(DFVF) DENMARK
Friedrich-Loeffler- Sdufer 10 Dr Bernd Haas bernd.haas@fli.bund.
Institut 17493 Greifswald- de
Bundesforschungs- Insel Riems
institut fr Tierge- GERMANY
sundheit (FLI)
National Institute Carretera de Algete Dr Esther Blanco blanco@inia.es
for Agriculture and a El Casar de
Food Research and Talamanca
Technology (INIA) 28130 Valdeolmos
SPAIN
Instituto Via Bianchi 7/9 Dr Emiliana Brocchi emiliana.brocchi@
Zooprofilattico Speri- 25124 Brescia bs.izs.it
mentale della Lom- ITALY
bardia e dellEmilia
Romagna (IZSLER)
Sap Institute (Sap) PO Box 714 Dr Fuat Ozyoruk fuato@sap.gov.tr
06044 Ulnus-Ankara
TURKEY
Agence 22 rue Pierre Curie Dr Stphan Zientara szientara@vet-alfort.fr
Franaise de Scurit 94703 Maisons-Alfort
(AFSSA) FRANCE
Institute Sensemattstrasse Dr Kenneth kenneth.mccullough@
of Virology and 293 McCullough ivi.admin.ch
Immunoprophylaxis 3147 Mittelhusern
(IVI) SWITZERLAND
[CA FMD/CSF]
Foot-and-mouth disease (FMD)

and classical swine fever
(CSF) coordination action
Acronym: Summary reappraisal of the stamping out approach,
CA FMD/CSF This coordination action gathered and a control method that is increasingly abhor-
shared information relevant to the control rent to the public. In the case of FMD, a new
Project number:
513755 of two of the most important OIE Notifiable EU directive places a greater emphasis on
diseases foot-and-mouth disease (FMD) the use of vaccination rather than slaughter
EC contribution:
and classical swine fever (CSF), both of to control future outbreaks, but this poses
EUR 1 400 000
which pose serious threats to our livestock veterinary and social problems.
Duration: industries. The project has strengthened
42 months
the networks of national reference labora- Ideally, better vaccines and better diagnos-
Start date: tories (RL) for these diseases by providing tic tests are needed, more information is
1 January 2005 coordination, web-based network resources, required on many aspects of vaccine per-
Instrument: disease management manuals and mech formance, better global surveillance and
Coordination action anisms. The involvement of the EC, OIE and risk analysis would help us to choose which
EUFMD/FAO has enhanced and expanded vaccines to stockpile, and new modelling
European networks with inputs from experts tools could help with the difficult decisions
in other countries. The project focused on as to which variant of a particular control
the coordination of research, global disease policy to use. Ultimately, the best way of
surveillance, risk analysis, vaccine reserves, protecting Europe against FMD and CSF is
diagnostics, laboratory preparedness, con- to control them elsewhere at their endemic
trol policies including vaccination and wild sources. In Turkey, where FMD is still
boar issues. It has initiated and strenthened endemic, vaccination is being used to help
the EU and International scientific networks bring about the eradication of the disease.
of FMD reference laboratories, vaccine There is pressure to stop CSF vaccination
banks, the Community Reference Labora- campaigns in several new Member States
tories for both FMD and CSF, harmonised as they are about to join the EU, but the dis-
disease control tools, fostered trust and ease is still endemic in the wild boar popu-
facilitated collaboration through informa- lations of some of these countries. Without
tion exchange and provided information to a more global approach, it is most unlikely
the wider stakeholder community. that these diseases can be controlled, let
alone eradicated. A global approach would
benefit both Europe and other countries and
Problem would build a framework for the control of
FMD and CSF are the subject of continuous other significant livestock diseases.
eradication efforts in Europe and other parts
of the world where fully susceptible, densely The critical mass of scientific effort
populated and highly mobile livestock popu- dedicated to improving the control of FMD
lation exist. This difficulty has led to the and CSF is small and highly fragmented
and, consequently, scientific expertise is reference laboratories that provides

highly dispersed. Each nation tends to sup- better coordination, cooperation and
port its own limited laboratory facilities collaboration across a variety of surveil-
without sufficient international interaction. lance and diagnostic activities between
Further afield, the same uncoordinated, and the European Commission and national
often shallow, effort may be duplicated in reference laboratories, and international
other parts of the world, whilst cutting- animal health |organisations.
edge research activities may be almost The formation and continuation of the
completely lacking or ignored by the official International Foot-and-Mouth Disease
bodies. Where expertise already exists on Vaccine Strategic Reserves Network that
these diseases, it is not readily accessible. provides a platform for a coordinated
It is obvious that this is not an efficient approach to antigen/vaccine bank activ
model for internationally effective use of ities around the world facilitating the
resources with regard to research to sup- harmonisation of standards for vaccine
port EU policy. Improved coordination of bank antigens, ensuring better control
FMD and CSF research and other activities of FMD in the event of an outbreak, and
is therefore urgently needed. reducing some of the costs attributing to
the maintenance of such reserves with a
concept of sharing resources.
Aim The project has helped to overcome
To strengthen and coordinate existing long-lasting problems in coordination of
initiatives for collaborative actions involving: CSF research as well as in harmonisa-
tion of diagnostic methods and disease
the Community Reference Laboratory/ control tools. It supported strategic plan-
National Reference Laboratories network; ning in the development of new vaccines
the Research Group of FAOs European and cooperation with another EU pro-
Commission for the Control of FMD; gramme, EPIZONE, the more effective
the network of reference laboratories of use of available resources.
the Office International des Epizooties; The project activated and boosted the
other international stakeholders involved collaboration between CSF reference
in FMD and CSF research. laboratories and promoted research
cooperation. It helped to develop
To initiate new collaborative action on: improved methods and techniques for
diagnostic. CSF diagnostics and vac-
research activities and needs; cine performance were assessed and
global sur veillance and risk published in a scientific article. In add
management/research; ition a workshop on Classical swine
diagnostic harmonisation and laboratory fever Assessment of control tools
preparedness; and research gap was held in Hannover
vaccine/antigen banks; in cooperation with the American Agri-
the problem of CSF in wild boar; cultural Research Service, United States.
refinement of disease management and As a result, the report Future needs and
control options. goals of CSF research was provided to
funding bodies.
Results An agreement was reached between the
The following are the key achievements of CSF reference laboratories on contem-
the project. porary CSF diagnostic techniques and
standards. The results were reviewed
The formation and strengthening of in the Diagnostic Manual establishing
the global network of OIE and FAO FMD diagnostic procedures, sampling
methods and criteria for evaluation of References/publications

the laboratory tests for the confirm Barnett, P.V., Bashiruddin, J.B., Hammond,
ation of classical swine fever. To ensure J.M., Geale, D., Paton, D.J. (2010), Toward
proper and effective diagnostic in cases a Global FMD Vaccine Bank Network, OIE
of emergency like an outbreak of CSF in Review 29, 593602.
fully susceptible and densely populated
livestock, guidelines for laboratory con- Blome, S., Meindl-Bhmer, A., Loeffen, W.,
tingency planning and real-time exer- Thuer, B., Moennig, V. (2006), Assessment
cises were established. They will help of classical swine fever (CSF) diagnostics
laboratories to develop their own con- and vaccine performance, Rev. sci. tech. Off.
tingency plans and facilitate real-time int. Epiz., 25, 10251038.
exercises.
The partners developed guidelines for Koenen, F., Uttenthal, A., Meindl-Bhmer,
the harmonisation and improvement of A. (2007), Real-time laboratory exercises
international comparison testing. There- to test contingency plans for classical
upon an advisory board for comparison swine fever: experiences from two national
tests was established. laboratories, Rev. sci. tech. Off. int. Epiz., 26,
The collaboration of researchers from 629638.
different countries reviewed control
tools, control strategies and risk ana Marshall, M.J., Roger, P.A., Bashiruddin, J.B.
lyses for CSF in pigs and wild boar and (2006), Making better use of technological
distributed their results in several scien- advances to meet stakeholder needs, Rev.
tific publications. During the workshop sci. tech. Off. int. Epiz., 25, 233251.
Refinement of disease management
and control options, experiences were Project website
exchanged between scientists, policy- http://www.foot-and-mouth.org/fmd-csf-ca
makers, representatives of the OIE and
FAO, retail organisations, and agrarian Keywords
associations on the pros and cons of foot-and-mouth disease, classical swine
the use of currently available (marker) fever, FMD, CSF, network, coordination,
vaccines in the eradication of CSF, more collaboration, cooperation
specifically the options for trade of vac-
cinated animals and their products. The Coordinator
conclusions and recommendations of Dr David J. Paton
this meeting were summarised and for- Institute for Animal Health
warded to the EU commission. Ash Road
The provision of a publicly open and Woking
interactive web-based disease aware- Surrey GU24 0NF
ness, communication and education UNITED KINGDOM
facility for FMD scientists and stake- E-mail: david.paton@bbsrc.ac.uk
holders (http://www.foot-and-mouth.
org/) and a restricted but interactive CSF
database for scientists (http://viro08.
tiho-hanover.de/eg/csf).
Open publications of guides and tem-
plates for the preparation of Laboratory
Contingency Planning documents for
FMD and CSF (available at Laboratory
Preparedness, http://www.foot-and-
mouth.org/fmd-csf-ca).
Partners
Institute of Virology Bunteweg 17 Prof. Dr Volker Volker.Moennig@tiho-
School of Veterinary 30559 Hannover Moennig hannover.de
Medicine GERMANY
Office International 12 rue de Prony Dr Gideon Bruckner g.bruckner@oie.int
des Epizooties 75017 Paris
FRANCE
Food and Agriculture Viale delle Terme di Dr Keith Sumption keith.sumption@fao.
Organisation Caracalla org
Animal Health Service 00100 Rome
Animal Production ITALY
and Health Division
Danish Technical 4771 Kalvehave Dr se Uttenthal AAU@vet.dtu.dk
University DENMARK
Section Epizootic Groeselenberg 99 Dr Kris De Clercq krdec@var.fgov.be
Diseases 1180 Ukkel
CODA-CERVA-VAR BELGIUM
Veterinary New Haw Dr Trevor Drew t.w.drew@vla.defra.gsi.
Laboratories Addlestone gov.uk
Agency Surrey KT15 3NB
UNITED KINGDOM
Institute of Sensemattstrasse 293 Dr Barbara Thuer Barbara.Thuer@ivi.
Virology and 3147 Mittelhaeusern admin.ch
Immunoprophylaxis SWITZERLAND
Centraal Instituut Houtribweg 39 Dr Willie Loeffen willie.loeffen@wur.nl
Dierziekte Controle 8203 AA Lelystad
Lelystad NETHERLANDS
Friedrich-Loeffler- Boddenblick 5a Dr Bernd Haas bernd.haas@rie.bfav.
Institute 17493 Greifswald- de
National Reference Insel Riems
Laboratory for FMD GERMANY
Agence Francaise de 22 rue Pierre Curie Dr Stphan Zientara s.zientara@afssa.fr
Securite Sanitaire des 94703 Maisons-Alfort
Aliments Alfort FRANCE
[FMD-Disconvac]
Development, enhancement
and complementation of
animal-sparing, foot-and-
mouth disease vaccine-based
control strategies for free
and endemic regions
Acronym: Summary generation FMD vaccines and diagnostics
FMD-Disconvac Foot-and-mouth disease (FMD) is one of by applying cutting-edge technologies; and
the most infectious diseases of livestock (iv) the enhancement of our knowledge on
Project number:
226556 and continues to pose a significant threat FMD spread and transmission following the
to endemic and free regions alike. The use of high-potency monovalent or multi-
EC contribution:
impact of FMD on society and international valent vaccines. The role of wildlife (buffalo,
EUR 2 998 863
trade is high, thereby demanding stringent gazelles and wild boar) in FMDV mainten
Duration: prevention, surveillance and control plans ance and transmission will also be inves-
39 months
taken up in crisis preparedness plans. On tigated. The project consists of seven dif-
Start date: the other hand, there is a global increased ferent, yet interlinked, work packages (WP)
1 April 2009 demand for animal welfare and ethical con- each addressing one of the items listed in
Instrument: siderations necessitating a decreased reli- the work programme topic KBBE-2008-1-3-
Collaborative project ance on eradication of animals to control 02, and led by renowned WP leaders with
FMD virus (FMV) spread, and on the use of years of relevant experience in the field of
animals for the regulatory testing of veter FMD. As such, significant progress towards
inary products. The project seeks to balance the objectives of the EUs Animal Health
these apparently contrasting viewpoints by Strategy (200713), the European Technol-
addressing specific gaps in our knowledge ogy Platform for Global Animal Health, and
on all aspects of FMD control to enable the Global Roadmap for improving the tools
implementation of enhanced animal- to control FMD in endemic settings will be
sparing vaccine-based control strategies achieved.
tailored to the needs of free and endemic
settings. Consequently, four main object
ives have been identified, including: (i) the Problem
improvement of the quality of existing FMD Although the EUs Animal Health Strat-
vaccines and diagnostics; (ii) the refinement egy (200713) recognises that significant
and replacement of in vivo FMD vaccine scientific and logistic advances in FMD
quality tests; (iii) the development of new prevention, diagnosis, surveillance and
control have been made in recent years have been identified towards which sci-
(e.g. through the sixth framework pro- entific and technological efforts will be
gramme projects FMD_ImproCon (SSPE- directed:
C T-20 03-5036 03), EPIZONE (FOOD-
CT-2006-016236) and the coordination WP2: Reduction and refinement of
action FMD & CSF (SSPE-CT-513755)), in vivo vaccine quality tests by in
some drawbacks regarding the imple- vitro methods
mentation of animal-sparing, FMD vac- This WP aims to replace the current in vivo
cine-based control strategies for free and Gold Standard tests for vaccine efficacy
endemic regions persist and need to be (potency), purity and safety, in light of the
tackled to improve food safety, as well as 3R principle, by validated in vitro laboratory
economic and animal welfare aspects. tests. More specifically by: (i) the determin
ation and validation of correlation models
between in vitro laboratory tests and in
Aim vivo protection based on experimental and
The proposed consortium seeks to build field data; (ii) the development of in vitro
on expertise gained in the FMD_ImproCon immunoassays to monitor vaccine purity
project (SSPE-CT-2003-503603) to address by the reduction of FMDV non-structural
the remaining gaps, as identified by ETP- proteins content during vaccine purification
GAH, in current knowledge and expertise and in the final vaccine; and (iii) the devel-
regarding the implementation of animal- opment of alternative methods to quantify
sparing, FMD vaccine-based control strate- the antigen payload content in the final
gies for free and endemic regions, thereby vaccine.
combating the disease at source.
WP3: Assessment and improvement
By building on existing knowledge and of heterologous protection by FMD
through project-generated data, the project vaccines
aims to: This WP aims to predict how well a vac-
cine will protect against a challenge virus
(i) improve the quality of existing FMD of another strain within the same sero-
vaccines and diagnostics; type. Comparisons will be made between
(ii) refine and replace in vivo vaccine quality results of cross-challenge and homologous
tests; challenge tests. Correlations will be made
(iii)
develop new generation FMD vac- between observed cross-protection, pre-
cines and diagnostics by cutting-edge dicted serology and amino acid homology
technology; of the respective virus capsids. The work
(iv) and to increase/enhance our knowledge will focus on serotypes O and A.
on FMDV spread and transmission fol-
lowing the use of high-potency mono- Moreover, r-value determination between
valent or multivalent vaccines in free vaccine strains and FMDV field isolates will
and endemic settings. be improved by harmonising test meth-
odologies and drafting guidelines for the
The role of wildlife (buffalo, gazelle and reliable selection of reagents to include in
wild boar) in FMDV maintenance and in vitro vaccine matching studies, thereby
transmission will also be investigated. avoiding future in vivo cross-protection
studies.
Expected results The depth of our knowledge and expertise

Apar t from the management tasks regarding vaccine spectrum coverage will
described in WP1, six priority work packages be increased as well.
WP4: Development of vaccines and susceptible species in the period shortly

alternatives (antivirals) with rapid after applying emergency vaccination, and
onset of immunity and based on to study transmission dynamics in real-time
safer production methods outbreak situations to set up early warning
This WP aims to increase our knowledge systems for FMDV penetration. The effect
by investigating approaches for reinforc- of vaccination in preventing FMD transmis-
ing the mucosal immune response in order sion through contact exposure to the virus
to prevent FMDV infection at the primary will be studied by carefully designed FMD
portal for virus entry. Methods will be transmission experiments. A newly devel-
evaluated to elicit and measure mucosal oped infection model will be used to study
immunity against FMDV in cattle. Ways to the ability of the Asian buffalo to transmit
stimulate innate (rapid) and adaptive (last- FMDV infection and to investigate the effi-
ing) mucosal immune responses will be cacy of vaccination to prevent this. The role
investigated, using novel delivery systems, of wildlife in FMDV maintenance and trans-
adjuvants and viral vectors. Other new gen- mission, and quantified knowledge on the
eration vaccines will also be developed and presence of FMDV in viral secretions and
efficacy tested, avoiding the need for virus excretions in different species will be stud-
culture, thereby making the production of ied as well.
FMD vaccines environmentally safer.
WP7: Development or adaptation of
Moreover, the use of potent and selective computerised FMD spread models
anti-FMDV antiviral compounds, that rapidly to optimise vaccination schemes
and completely prevent FMDV replication, This WP aims to study the applicability and
will be investigated in order to decrease the feasibility of modifying existing simula-
post vaccination immunity gap. tion models (InterSpread Plus model, Davis
model, NAADS model and other models
WP5: Improvement in FMD within the consortium) for FMD spread to
diagnostics suit the exploration of vaccination strate-
This WP aims to: (i) increase the availability gies in the EU and other Western European
of FMD diagnostics; (ii) improve standard countries where FMD is considered an
isation and harmonisation of FMD diag- exotic threat.
nostic results; and (iii) develop new and,
possibly better, diagnostic tools for con- Computer models developed to enable vac-
firmatory tests and/or test systems for cination strategies to be designed for high-
NSP-serology. Therefore, a panel of sta- risk regions within countries belonging to
bilised, validated and reliable diagnostic this consortium, would be relevant stepping
kits for FMD serology and antigen typing, stones to model vaccination strategies for
ready for commercial exploitation, will be truly endemic regions of the world.
developed and/or validated (i.e. confirma-
tory NSP test, IgA in saliva ELISA, assays in
which serum reaction profiles are obtained Potential applications
simultaneously against a number of anti- The reduction and refinement of in vivo vac-
gens multiplexing). cine quality tests by in vitro methods will
guarantee the overall quality of the vaccine
WP6: Improving knowledge on batch in a verifiable form to end-users and
FMDV transmission between species other stakeholders, strengthening the pos
and in recently vaccinated animals ition of the EU on the global market when
This WP aims to obtain previously unavail- it comes to the implementation of the vac-
able quantified knowledge on FMDV trans- cinate-to-live policy. Consequently, reliance
mission within and between different FMDV on animals for regulatory testing of vaccine
batch release control will be decreased and References/publications

animal welfare increased (WP2). Lefebvre, D.J., Neyts, J., De Clercq, K. (2010),
Development of a foot-and-mouth disease
The assessment and improvement of heter- infection model in severe combined immu-
ologous protection by FMD vaccines will help nodeficient mice for the preliminary evalu-
decision-makers in their difficult choice of ation of antiviral drugs, Transboundary
which vaccine to use in future outbreaks and and Emerging Diseases, December2010,
in their responsibility to update and reinforce Vol.57 (6), pp. 430433.
FMD vaccine/antigen banks (WP3).
Nagendrakumar, S.B., Srinivasan, V.A.,
The development of new generation vaccines Madhanmohan, M., Yuvaraj, S., Parida,
and antiviral compounds, based on safe S., Di Nardo, A., Horsingston, J., Paton,
production methods and specifically aimed D.J. (2011), Evaluation of cross-protec-
to reduce the immunity gap shortly after tion between O1 Manisa and O1 Cam-
vaccination, will (i) supplement the existing pos incattle vaccinated with differ-
control tools to combat FMD and (ii) allow ent payloads of O1 Manisa monovalent
the enhancement of emergency contingency vaccine, Vaccine, 24 Feb., Vol. 29 (10),
plans enabling a better, quicker and animal- pp.19061912.
sparing response to FMD outbreaks (WP4).
Willems, T., Lefebvre, D.J., Neyts, J., De
The improvement in FMD diagnostics will Clercq, K. (2011), Diagnostic performance
help the OIE/EU to better interpret the dos and application of two commercially avail-
siers submitted to demonstrate/substanti- able cell viability assays in foot-and-mouth
ate FMD-freedom and, by facilitating and disease research, Journal of Virological
accelerating the development and distribu- Methods, Vol. 173, pp. 108114.
tion of the most effective diagnostics for
FMD in Europe and the rest of the world, 2010, project results were presented at
could contribute to the vision of the ETPGAH. the open session meeting of the EuFMD
Knowledge on performance characteristics standing technical committee, 27 Septem-
of available DIVA (differentiation of infected ber1 October, Vienna, Austria (http://www.
from vaccinated animals) diagnostics on a fao.org/ag/againfo/commissions/en/eufmd/
global scale will help to understand the FMD eufmd.html).
situation in other regions of the world, result-
ing in an increased awareness of the poten- Project website
tial threats to the European Union (WP5). http://www.fmddisconvac.net
Knowledge on FMDV transmission between Keywords

species and in recently vaccinated animals foot-and-mouth disease, disease con-
can be used to adapt and improve com- trol, vaccines, diagnostics, transmission,
puterised FMD spread models to optimise vaccination strategies
FMD vaccination programmes in free and
endemic settings alike (WP6). Coordinator
Dr Kris De Clercq
Computerised FMD spread models devel- Centrum voor Onderzoek in Diergenees
oped within this project could enable the kunde en Agrochemie
design of vaccination strategies for high-risk CODA
regions within countries belonging to this Groeselenberg 99
consortium and could be relevant stepping 1180 Ukkel
stones to model vaccination strategies for BELGIUM
truly endemic regions of the world (WP7). krdec@var.fgov.be
Partners
Institute for Ash Road Dr David Paton david.paton@bbsrc.
Animal Health (IAH) Pirbright, Guildford ac.uk
GU24 ONF
UNITED KINGDOM
Central Veterinary Houtribweg 39 Dr Aldo Dekker aldo.dekker@wur.nl
Institute of 8221 RA Lelystad
Wageningen UR (CVI) NETHERLANDS
Instituto Via Bianchi 9 Dr Emiliana Brocchi emiliana.brocchi@
Zooprofilattico Speri- 25124 Brescia bs.izs.it
mentale della Lom- ITALY
bardia e dellEmilia
Romagna (IZSLER)
Friedrich-Loeffler- Boddenblick 5A Dr Bernd Haas bernd.haas@fli.bund.
Institut 17493 Greifswald- de
Bundesforschungs- Insel Riems
institut fr GERMANY
Tiergesundheit (FLI)
Indian Immuno- Rakshapuram, Dr Viluppanoor srini@indimmune.com
logicals Limited (IIL) Gachibowli Srinivasan
Hyderabad 500 032
INDIA
Lanzhou Xujiaping 1 Dr Li Pan Panlizhe@yahoo.com.
Veterinary Research CHN 730046 Lan- cn
Institute (LVRI) zhou, Gansu
CHINA
Fundacin para la Los Reseros y las Dr Mariano Prez mperez@cnia.inta.
interaccin de los Cabaas Filgueira gov.ar
sistemas 1686 Hurlingham,
productuvo, educativo, prov. Buenos Aires
cientfico-tecnolgico ARGENTINA
(FUNPRECIT)
Agence Avenue du Gnral de Dr Stphan Zientara szientara@vet-alfort.fr
Franaise de Scurit Gaulle
Sanitaire des Aliments 94703 Maisons-Alfort
(AFSSA) FRANCE
Institute Sensemattstrasse Dr Kenneth kenneth.mccullough@
of Virology and 293 McCullough ivi.admin.ch
Immunoprophylaxis 3147 Mittelhusern
(IVI) SWITZERLAND
Ministry of PO Box 12 Dr Hagai Yadin hagaiy@moag.gov.il
Agriculture, Kimron Beit-Dagan 50250
Veterinary Institute ISRAEL
(KVI)
University of University Avenue Prof. Daniel Haydon D.Haydon@bio.gla.
Glasgow (UGLA) Glasgow G12 8QQ ac.uk
UNITED KINGDOM
National Veterinary Blowsvej 27 Dr Claes Ene cen@vet.dtu.dk
Institute, Danish 1790 Copenhagen
Technical University DENMARK
(DTU)

Merial SAS 29 Avenue Tony Dr Philippe Dubourget Philippe.DUBOURGET@
Garnier merial.com
69348 Lyon
FRANCE
2.2. Classic swine fever

[CSFVACCINE & WILD BOAR]
Epidemiology and control

of classical swine fever (CSF)
in wild boar and potential
use of a newly developed
live marker vaccine
Summary boar density is recognised as one of the
Acronym:
Classical swine fever virus (CSFV) is a most relevant risk factor enabling endemic CSFVACCINE &
recurring infection of domestic pigs and infections. Direct and indirect contacts WILD BOAR
wild boar. Notwithstanding the relative between domestic and wild species and
Project number:
success in eradicating the disease in cer- illegal swill feeding practices have been 501599
tain parts of Europe and despite intensive demonstrated as the main source for the
EC contribution:
efforts on a national as well as on an inter- spreading of the infection in domestic pigs.
EUR 1 656 760
national level, the complete eradication The relevant risk of introduction of CSFV
of CSF in Europe has proved to be elusive. from wild boar is clearly demonstrated by Duration:
4 years and 9 months
Additionally, the disease is still endemic the facts that about 80 % of primary CSF
in some new EU Member States (Bulgaria outbreaks in domestic pigs have occurred in Start date:
and Romania) and sporadic outbreaks still regions where CSF in wild boar is endemic. 1 January 2004
occur in free areas. Although the specific Therefore, any programme aimed at a Instrument:
role of wild boars as CSFV reservoirs needs permanent eradication or control at long specific targeted
to be further clarified, the CSFV infection term of CSF should include methodologies research or innovation
project (STREP)
can persist in wild boar populations and is which allow not only an efficient approach
therefore probably responsible for CSF re- in domestic animals but also in wild boar.
emergence. The latter is very important as However, the latter was hampered by the
in the last decade, the wild boar population lack of knowledge regarding wild boar pop-
has increased all over Europe and wild ulation dynamics and structure as well as
epidemiological parameters influencing the infection in wild boar populations. Vaccin

course of the infection within it. By using ation can provide an alternative for this
existing literature, data from previous out- current strategy. However, the contempo-
breaks (such as Varese (Italy), Luxembourg rary available live vaccines such as C-strain,
and Mecklenburg-Western Pomerania (Ger- although potent and oral applicable, do
many)) or the development of new applica- not allow serological differentiation from
tions, different parameters were obtained infected animals. In contrast, such adif-
and subsequently used to design a new ferentiation is possible with the commer-
mathematical model based on a metapopu- cial E2-marker subunit vaccine. However,
lation principle. The mathematical develop- this commercial E2-marker subunit vac-
ment of this model was validated using field cine needs to be administrated parenterally
data for outbreaks in Italy and Switzerland. as oral application is not possible and can
The major benefit of this model is that it therefore not be used for CSFV manage-
allows a better understanding of CSFV epi- ment in wild boar. Due to the shortcomings
demiology in wild boar populations and the of the current available E2-marker subunit
evaluation of the impact of control strate- vaccine there is a need for a new type of
gies (such as hunting and vaccination) on vaccine which is not only efficient but also
the course of the infection. Hunting showed has DIVA (differentiation of infected from
to be an ineffective way to control the vaccinated animals) capacity and can be
infection as only unrealistica lly intensive administrated orally. This was addressed
hunting efforts (> 70 %/year) could eradi- during this project by the development of a
cate the infection. Although in small popu- live marker vaccine, whereby the E2-region
lations (< 1000 animals) a non-intervention of BVDV (CP7) and CSFV (C-strain) were
policy revealed to be successful, vaccination replaced by the corresponding sequence
was demonstrated to be an effective tool in of CSFV (Alfort 187) and BDV (Gifhorn)
controlling the CSFV infection as it always respectively. A select number of chimeri-
reduces the epidemic peak. The chance of cal candidates, displaying the desired in
a successful eradication of the infection vitro characteristics (such as replication
is determined by the percentage of the kinetics, cell specificity and DIVA potential),
susceptible population that is vaccinated were subjected to extensive in vivo evalu-
within a short range of time (approximately ation. Although the CSFV-based candidates
15 days). While a 60 % vaccination rate of (e.g. pRiems_ABCgif) were safe to use, as
the susceptible animals will lead to prompt no adverse effects were noted in the inocu-
eradication, 20 % will increase the probabil- lated animals irrespective of the method of
ity of endemic stability of the infection. The application, the protection was not always
latter clearly shows that when the decision complete, especially when given orally, or
is made to vaccinate, it should be done in differentiation with wild type CSFV was not
such a manner that a very large portion of unequivocally possible using commercial
the population is reached. ELISA systems. Conversely, CP7_E2alf was
found to protect domestic pigs as well as
On the domestic pig level, the current con- wild boar completely against a lethal CSFV
trol and eradication strategies for CSF man- challenge when given intramuscularly, oro-
agement are based on stamping out the nasally or by the newly developed baits. This
infection combined with pre-emptive cull- complete protection was already achieved
ing. Notwithstanding the efficacy of this after 7 and 14 days following intramuscular
approach, there is an increasing ethical or oronasal vaccination respectively. Initial
concern and public resistance, as numerous potency studies have indicated that a dose
healthy, uninfected animals are killed and of 103.5 10 4.5 TCID50 is sufficient for clinical
destroyed. Furthermore, this stamping-out protection against a lethal challenge when
policy is not suited for management of the administrated intramuscularly, with 100 %
of the animals reaching a protective VNT of bovine to porcine previously observed in

1:20 at 21dpv. Using oronasal application, vitro. The field applications during CSFV
the potency of CP7_E2alf was similarly vaccination campaigns in wild boar have
confirmed and was found to be compara- encountered another problem, namely the
ble to that of a commercial C-strain vac- limited uptake of baits by young animals.
cine. Furthermore, the immunity granted by The latter is important because indepen-
CP7_E2alf was sterile, as no evidence was dently of the efficacy of the vaccine, this
found of shedding, excretion or transmis- can lead to a part of the wild boar popula-
sion to contact animals in any of the ani- tion, already more vulnerable to infection,
mals experiments performed, and could remaining unprotected. This could contrib-
be achieved at a dose of approximately 2 ute to the persistence of the infection. For
10 5.5 TCID50. Furthermore, a long-lasting this purpose new small spherical and cuboid
antibody response was observed at least baits were designed and constructed. The
up to 98 days dpv, even when a suboptimal new 3 cm spherical bait clearly showed an
dose was used of 2 10 4.5 TCID50. The lat- improved uptake rate in young animals up
ter is important as such a dose may occur to three-and-ahalf months. However, even
when there is incomplete bait uptake or this new small bait was not taken up by
where vaccine titres are reduced in the field animals younger than three months, prob-
or during the production process. CP7_E2alf ably due to the fact that they prefer suck-
is not only highly efficient, it is also safe ling. This has important implications in any
to use. The latter was demonstrated as in vaccination strategy as it has to be kept in
none of the performed animal trials were mind that these young animals cannot be
any significant adverse effects observed immunised in this way. Repeated baiting
in domestic pigs and wild boar. Similarly, or baiting at a time point sufficiently long
no effects on farrowing or the birth per- after farrowing (when all the gruntlings are
formance of the piglets were observed on older than three months) is therefore desir-
vaccination of pregnant sows and no evi- able. In summary, based on in vitro and in
dence of transplacental transmission was vivo results, it can be clearly stated that
found. One of the goals of this project was the CP7_E2alf is the most suited vaccine
to develop a vaccine that can be used for candidate coming forth from the used dual
CSF management in wild boar. A number approach and the resulting numerous chi-
of specific safety issues are linked to the merical constructs. The great benefit, how-
field application of a vaccine, such as the ever, of the used strategy is that not only
transmission of the vaccine to other spe- a very good candidate has been obtained
cies. This is even more important due to the but that a backup is available, which is
BVDV-background of CP7_E2alf. As previ- advantageous if at a later stage problems
ously stated, the immunity achieved by should occur with the CP7_E2alf, and that
CP7_E2alf is sterile (no shedding, no excre- additional constructs are at hand which can
tion and no contact transmission), meaning be used to study the different aspects of
that transmission to other species by animal live marker vaccines, such safety features,
to animal contacts is very unlikely. Indirect growth and immunogenic enhancers and
transmission, by uptake of the baits (filled soon.
with vaccine) is equally unlikely because
no clinical, serological or virological evi- Following the development and testing of
dence could be found for the presence and the vaccine candidates, production on an
shedding of the CP7_E2alf in young rumi- industrial scale of the final vaccine candi-
nants and rabbits on oral application, and date CP7_E2alf and the backup candidate
even after intramuscular application to pRiems_ ABC_gif was established and
cattle and sheep. The latter confirmed the resulted in a master and working cell
changed cell tropism of CP7_E2alf from stock of the sk-6 cell line and a master
and working seed virus. All the stocks were techniques combine high throughput capac-
prepared in full compliance with European ity with a superior sensitivity and reduced
Pharmacopoeia guidelines regarding ster labour requirements, compared to virus iso-
ility and freedom of extraneous contamin lation (VI). Such advantages are important
ants. In order to be able to use as much during outbreaks because they allow an ear-
as possible of the data generated dur- lier detection of the infection (in this project,
ing the animal trials for future licensing/ real-time RT-PCR detected CSFV one to two
registration, 125 vials (volume: 20 ml per days earlier than VI), and permit a larger
vial) at 10 6.4 TCID50/ml of the pilot vaccine number of samples to be analysed within
CP7_E2alf were produced following good a shorter time. The latter is further accen-
manufacturing practice (GMP) and the tuated by the ability to pool at least five
requirements of European Pharmacopoeia samples for real-time RT-PCR evaluation
requirements regarding sterility. without loss of sensitivity as demonstrated
in this project. However, early detection is
Simultaneously with the development of influenced by the choice of tissue used for
a new vaccine, the necessary serologi- analysis. The detection of the virus in ton-
cal and virological diagnostic tools were sil preceded that of other tissues, including
developed and subsequently evaluated in blood, and is therefore the most suited tis-
an interlaboratory evaluation (ILE). At the sue for early as well as long-term detection.
start of the project, it was decided to evalu- On the other hand, blood can be used as an
ate the DIVA potential of existing commer- alternative at herd level if a larger number
cial ELISA systems. Similar to earlier initial of samples are analysed to compensate for
results, differentiation between vaccinated the lower probability of detecting the virus.
and infected animals was possible for CP7_ The availability of accompanying genetic
E2alf and, to a lesser degree, for the CSFV- diagnostic tools is an important criterion
based candidate, using existing commercial for a more generalised use of a CSFV vac-
systems (E RNS -antibody ELSIA). However, cine. Therefore, classic gel-based and real-
diagnostic sensitivity and specificity should time RT-PCRs were developed that are able
be improved and batch-to-batch conform- to differentiate between CSFV and the new
ity should be addressed as variable results vaccine candidate CP7_E2alf (genetic DIVA).
were obtained with different batches. Not- As expected, the real-time RT-PCR has a
withstanding the fact that the focus was slightly higher sensitivity compared to the
placed on the use of existing ELISA sys- classical systems with a detection limit of
tems, the potential of creating adapted 50 (CSFV) and 500 copies (CP7_E2alf) of
versions or the development of new ELISAs cRNA per reaction for the classical system
was analysed by searching for Mabs with and 20 (CSFV) and 50 (CP7_E2alf) copies
DIVA potential. The latter not only resulted of synthetic cRNA for the real-time RT-PCR.
in an increased insight into the epitopic Although single-tube formats have been
structure of E2 and E RNS proteins of both developed for both, the two-tube format
CSFV and BVDV, which will be beneficial is preferable as its sensitivity is 10 to 100
for better understanding the immunogenic times higher. The applicability and DIVA
characteristics of vaccines, but also in the capacity of the newly developed real-time
identification of multiple Mabs that can be RT-PCR was confirmed in the interlaboratory
used for the development of DIVA ELISAs evaluation where it was found to be func-
for either CP7_E2alf or pRiems_ABC_gif tioning with very good robustness and
reproducibility, although some optimisation
During the last decade, PCR-based tech- and additional testing is required for the
niques have become more and more import CP7_E2alf detection. Despite the diagnos-
ant as a diagnostic tool, especially with the tic potential of PCR-based methods, some
development of real-time RT-PCR. These issues have arisen regarding samples which
were scored positive by PCR but negative by development of a marker vaccine and
VI. Although research in the field of CSFV accompanying diagnostic assays and
was one of the first to report this problem, protocols.
it is not restricted to it as it is now also
described for other viruses. In addition to Results
differences in detection sensitivity and anti- Development of an epidemiological
body complexation, it was demonstrated, by model for CSF eradication in wild
using a newly developed RT-PCR panel, that boar
the presence of viral genome fragments in A number of epidemiological parameters
the sample can equally be the cause of dis- were either determined by using existing
crepant PCR and VI results. literature, data collection or by develop-
ing new applications for estimating them
Not only DIVA diagnostic tools to be used (such as transmission coefficient and the
in combination with the new vaccine candi- minimum wild boar number). For the latter,
date have been developed during this pro- a user-friendly Microsoft Office Excel sheet
ject but also on-site or field detection tools. was developed based on the hunting bag.
The latter further enhances the early detec- Subsequently, a new mathematical model
tion of CSFV. A new lateral flow detection based on a meta-population principle was
prototype, incorporating a specific CSFV designed and validated using data from
Mab, has been developed following numer- previous outbreaks. This new model showed
ous screenings. Although further specific- that hunting is an ineffective way to control
ity and sensitivity testing is required, the the infection as only unrealistically inten-
clear and specific signal obtained clearly sive hunting efforts could eradicate the
underlines its promising potential. infection. Although in small populations
(<1000 to 1 500 animals) a non-interven-
In summary, the work performed during tion policy revealed to be successful, vac-
this CSFVACCINE & WILD BOAR resulted cination was demonstrated to be an effec-
not only in an enhanced insight into the tive tool in controlling the CSFV infection
epidemiological situation of CSF in wild as it always reduces the epidemic peak.
boar but also to a validated model allowing The chance of successful eradication of the
the evaluation of the impact of different infection is determined by the percentage
control strategies on the infection course. of the susceptible population that is vac-
In addition, with the development and the cinated within a short range of time. While
production under GMP conditions of a novel a 60 % vaccination rate of the susceptible
potent and safe live marker vaccine and animals will lead to prompt eradication, 20
its accompanying genetic and serological % will increase the probability of endemic
DIVA tools, an interesting alternative can stability of theinfection.
be presented to the current control and
eradication strategies. Adaptation of the C-strain vaccine
baits for use in wild boar with
special attention to young animals
Aim New small spherical and cuboid baits were
The main objectives of the project were the: designed and constructed. The new 3 cm
spherical bait clearly showed an improved
development of an epidemiological and uptake rate in young animals up to three-
economic model for CSFV eradication in and-a-half months. However, even this
wild boar; new small bait was not taken up by ani-
adaptation of the C-strain vaccine mals younger than three months, probably
baits for use in wild boar with special due to the fact that they prefer suckling.
attention to young animals; This has important implications in any
vaccination strategy as it has to be kept in European excellence in strategies to pre-

mind that these young animals cannot be vent or limit the introduction of an exotic
immunised in this way. Furthermore, it was agent (OIE List A) and to avoid transmission
demonstrated that lyophilization increased of pathogens between species (wildlife to
vaccine stability under field conditions farmed animals). It will reinforce consider-
and is therefore a promising method to ably the competitiveness of the European
increase bioavailability during vaccination pig production. By developing a vaccine with
campaigns. accompanied diagnostic assays the societal
problem of mass killing of healthy animals
Development of a marker vaccine can also be reduced.
and accompanying diagnostic
assays and protocols
A new live marker vaccine was developed References/publications
whereby the E2-region of BVDV (CP7) was Beer, M., Reimann, I., Hoffmann, B., Depner,
replaced by the corresponding sequence of K. (2006), Novel marker vaccines against
CSFV (Alfort 187). Based on in vitro and in classical swine fever, Vaccine, 6: 25(30):
vivo results, it can be clearly stated that 566570.
CP7_E2alf is the most suited vaccine candi-
date as it not only provides complete sterile Knig, P., Lange, E., Reimann, I., Beer, M.
immunity, independently form the applica- (2007), CP7_E2alf: A safe and efficient
tion method, it is also very safe to use. In marker vaccine strain for oral immunisation
none of the animal experiments were any of wild boar against classical swine fever
adverse effects noted on health or farrow- virus (CSFV), Vaccine, 30: 25(17): 33919.
ing with normal birth performance of the
piglets. Notwithstanding the BVDV back- Liu, L., Widn, F., Baule, C., Belk, S. (2007),
ground of CP7_E2alf, no serological or A one-step, gel-based RT-PCR assay with
virological evidence could be found for the comparable performance to real-time RT-
presence of the CP7_E2alf in young rumi- PCR for detection of classical swine fever
nants and rabbits on oral application, and virus, Journal of Virological Methods, (139):
even intramuscular application to cattle 20320.
and sheep did not result in detectable vac-
cine virus replication or shedding. In addi- Rasmussen, T.B., Uttenthal, A., Reimann, I.,
tion to the safety of this candidate, animals Nielsen, J., Depner, K., Beer, M., Virulence,
immunised with CP7_E2alf can be differ- immunogenicity and vaccine properties
entiated from wild-type infected animals of a novel chimeric pestivirus, Journal of
using either commercial ELISA system or by General Virology, February 2007, 88(Pt 2):
a real-time RT-PCR developed during this 4816.
project. The robustness of the developed
real-time RT PCR was confirmed during an Tignon, M., Kulcsr, G., Belk, K., Haegeman,
interlaboratory evaluation. A., Barna, T., Fbin, K., Lvai, R., Farsang,
A., Van der Stede, Y., Vrancken, R., Koenen,
F. (2008), Application of a Commercial
Potential applications Real-Time RT-PCR Assay for Surveillance of
The development of an epidemiological Classical Swine Fever: Evaluation by Testing
and economic model for CSF eradication in Sequential Tissue and Blood Samples, The
wild boar in combination with the develop- Open Veterinary Science Journal, 2008, 2,
ment of a live marker vaccine and accom- 104110.
panying diagnostic assays, to adapt this
vaccine for use in wild boar with special Project website
attention to young animals will enhance http://www.csfvaccine.org
Keywords Coordinator
classical swine fever, wild boar, live marker Frank Koenen
vaccine, DIVA, epidemiology, diagnostics Veterinary and Agrochemical Research
Centre
(CODA-CERVA-VAR)
Groeselenberg 99
1180 Ukkel
BELGIUM
frank.koenen@var.fgov.be
Partners
ORGANISATION ADDRESS CONTACT
The National Veterinary Box 585, Biomedical Center Sandor.Belak@sva.se
Institute (NVI) SE-751 23 Uppsala
Department of Virology SWEDEN
The National Veterinary Uppsala Katinka.Belak@sva.se
Institute (NVI) SWEDEN
Department of Pathology
Friedrich-Loeffler-Institut (FLI) Boddenblick 5a Martin.Beer@fli.bund.de
Institute of Diagnostic 17493 Greifswald-Insel Riems klaus.depner@fli.bund.de
Virology GERMANY Volker.Kaden@fli.bund.de
Friedrich-Loeffler-Institut (FLI) Seestrae 55 Mathias.kramer@fli.bund.de
Institute of Epidemiology 16868 Wusterhausen Christoph.Staubach@fli.bund.de
GERMANY
School of Veterinary Medicine Bnteweg 17 Volker.Moennig@tiho-hanno-
Hannover (TiHo) 30559 Hannover ver.de
Institute of Virology GERMANY
Istituto Nazionale per la Via Ca Fornacetta 9 infsvete@iperbole.bologna.it
Fauna Selvatica (INFS) 40064 Ozzano Emilia
ITALY
Institute of Virology and 3147 Mittelhaeusern Martin.Hofmann@ivi.admin.ch
Immunoprophylaxis (IVI) SWITZERLAND
Diagnostics Department
Fort Dodge 17813 Vall de Bianya (Girona) planaj@md.ahp.com
Veterinaria SA SPAIN
(FDV)
Universidad Avda. Puerta de Hierro s/n jmvizcaino@vet.ucm.es
Complutense 28040 Madrid
de Madrid (UCM) SPAIN
Department Sanidad Animal,
Facultad de Veterinaria
Central Agricultural Office, 10 PO Box 318 kulcsarg@oai.hu
Directorate of Budapest
Veterinary Medicinal Products 1475
HUNGARY
[CSF-GoDIVA]
Improve tools and strategies

for the prevention and
control of classical swine fever
Acronym: Summary population in most EU countries. Never-
CSF-GoDIVA Although classical swine fever (CSF) has theless, and despite intensive efforts at
Project number: been eradicated in wide areas within the EU, national and international level, sporadic
227003 the disease is endemic in some new Member outbreaks still occur in areas which are
EC contribution: States particularly in backyard pigs. In order free of CSF. Additionally, the disease is still
EUR 2 999 983 to improve eradication strategies, the project endemic in some countries that recently
Duration: aims are: (a) the final development and test- joined the EU Bulgaria and Romania; CSFV
48 months ing of a live marker vaccine candidate for the is widespread among wild boar populations
prevention and improved control of CSF, both on the European continent. Furthermore,
Start date:
1 March 2009 orally and intramuscularly applicable; (b) the CSFV is present in the backyard pig popu-
development and optimisation of accom- lation in Bulgaria and especially Romania.
Instrument:
panying discriminatory diagnostic tests; The epidemiology of CSF in this type of pig
Collaborative Project
(c)the production of an effective, oral deliv- holding which is novel within the European
ery system for the marker vaccine for use Union, has not yet been fully investigated.
in wild boar and backyard pigs; (d) the easy Therefore, a strong need exists to improve
selection of diseased animals. The improved knowledge and intervention strategies for
knowledge on immunological reactions and backyard pigs.
pathogenesis will support a more efficient
vaccine application and provide data for Currently, a CSF non-vaccination policy,
the epidemiological models. Epidemiologi- exists within the EU. However, due to prob-
cal studies of CSF in domestic and backyard lems with CSF in wild boar and in domestic
pigs and in wild boar including molecular pigs in some Member States, derogations
epidemiology intend to increase insight into from this non-vaccination policy have been
CSF transmission and persistence. Epidemio- granted and vaccination is applied in certain
logical models will be developed to support areas of Europe. For the oral vaccination of
risk assessment both for conventional eradi- wild boar, the modified live C-strain vaccine
cation strategies as well as for new strat- is used. This vaccine is very efficacious under
egies using the new vaccine and diagnostic field conditions but its major disadvantage is
tools including the role of CSF reservoirs. that no differentiation can be made between
The results concerning antiviral treatment vaccinated and infected animals. Baculovi-
will be evaluated and compared with the rus produced E2 subunit vaccines are avail-
traditional eradication strategies. able but data suggest that these vaccines
are less efficacious than conventional modi-
fied live vaccines and multiple vaccinations
Problem are needed before protection is obtained.
CSF is notifiable in the EU and has Moreover, subunit vaccines are not suit-
been eradicated from the domestic pig able to be used in baits for oral vaccination
of wild boar. The control of CSF outbreaks (viii) new models for the evaluation of the
in domestic pigs as well as in wild boar wild boar population and baiting;
and backyard herds would be significantly (ix) proof of principle for the use and
enhanced if asafe and efficacious marker registration of antiviral treatment.
vaccine with an accompanying DIVA (differ-
entiation of infected from vaccinated ani-
mals) diagnostic assay would be available.
Aim
The main goal of this project plan is com-
plex and multidisciplinary: (a) the final
development and testing of a live marker
vaccine (LMAV) candidate for the preven-
tion and improved control of classical swine
fever (CSF), both orally and intramuscularly
applicable; (b) the development and opti-
misation of accompanying discriminatory
diagnostic tests; (c) the production of an
effective, oral application system for the
marker vaccine for use in wild boar and
backyard pigs; (d) the easy selection of dis-
eased animals; (e) epidemiological studies The following is a summary of the results
of CSF in domestic and backyard pigs and achieved so far.
in wild boar, including molecular epidemi
ology and the study of alternative methods (i) The final live marker vaccine candidate
of suppression of viral replication. for intramuscular and oral application
was selected among two chimeric
pestivirus vaccine candidates.
Results (ii) A commercially available CSFV antibody
The expected results of this project are: ELISA was identified for use in conjunc-
tion with the new marker vaccine. New
(i) availability of new standardised diag- antigens and monoclonal antibodies for
nostic methods applicable to domestic the development of alternative DIVA
and backyard pigs and wild boar; ELISAs were also characterised.
(ii) validated new methods for the easy (iii) New small spherical baits were pro-
selection of suspicious animals; duced and their uptake evaluated in a
(iii) a better risk analysis including field study.
molecular epidemiology; (iv) An existing CSFV spread model for wild
(iv) a marketable third generation live boar was further developed to assess
marker vaccine for intra muscular and/ the impact of oral mass vaccination
or oral application in domestic and of wild boar and to assess the effect
backyard pigs and wild boar; of landscape structure and population
(v) new methods for the easy oral dynamics including hunting on the size
application of the vaccine; of outbreaks and on virus persistence.
(vi) better insight into the role and (v) A back yar d pig dat a ba se wa s
pathogenesis of CSF virus reservoirs; developed for Bulgaria.
(vii) new epidemiological models for CSF (vi) For domestic pigs, a management-
in domestic and backyard pigs and in orientated epidemiological model
wild boar; was developed as a tool to evaluate
the relative performance of differ- boar in Germany, The Journal of General

ent emergency control strategies in Virology, 91, 26872697.
domestic pigs.
(vii) The current surveillance strategies for Leifer, I., Lange, E., Reimann, I., Blome, S.,
CSF in wild boar and in domestic pigs Juanola, S., Duran, J.P., Beer, M. (2009),
were reviewed. Modified live marker vaccine candidate
(viii) The potential of an antiviral molecule CP7_E2alf provides early onset of protec-
to reduce the transmission of CSFV tion against lethal challenge infection with
from treated to untreated pigs was classical swine fever virus after both intra-
demonstrated. muscular and oral immunisation, Vaccine,
27, (47), 65229.
The combination of the final development Leifer, I., Depner, K., Blome, S., Le Potier,
and validation of a third generation LMAV M.-F., Le Dimna, M., Beer, M., Hoffmann, B.
with the discriminatory tests, the develop- (2009), Differentiation of C-strain Riems
ment of simple pen-side tests and the epi- or CP7_E2alf vaccinated animals from ani-
demiological evaluation of CSF in both non- mals infected by classical swine fever virus
vaccinated and vaccinated domestic pigs, field strains using real-time RT-PCR, Journal
backyard pigs and wild boars will result in of Virological Methods, 158 (12), 114122.
a better understanding and better tools in
order to fight this highly devastating dis- Rossi, S., Pol, F., Forot, B., Masse-provin,
ease. Improved immunisation and baiting N., Rigaux, S., Bronner, A., Le Potier, M.-F.
procedures as well as new LMAV and new (2010), Preventive vaccination contributes
procedures (DIVA, easy selection, sampling) to control classical swine fever in wild boar
should allow the efficient immunisation of (Sus scrofa sp.), Veterinary Microbiology,
pigs, and should enable an improved diagno- 142, (12), 99107.
sis including a discrimination of vaccinated
from infected animals (marker vaccines Project website
principle). The new available knowledge http://www.csfvaccine.org
concerning CSF epidemiology in particular
in backyard pigs, risk assessment, molecu- Keywords
lar epidemiology, control, immune response classical swine fever, live marker vaccine,
as well as the new diagnostic methods will antivirals, backyard pigs, wild boar, DIVA
help the animal health authorities inside diagnostics, epidemiology, thermography,
and outside the EU in their policymaking. molecular diagnosis, RT-PCR
Coordinator
References/publications Frank Koenen
Blome, S., Grotha, I., Moennig, V., Greiser- Veterinary and Agrochemical Research Centre
Wilke, I. (2010), Classical swine fever virus (CODA-CERVA-VAR)
in south-eastern Europe Retrospec- Groeselenberg 99
tive analysis of the disease situation and 1180 Ukkel
molecular epidemiology, Veterinary Micro- BELGIUM
biology, 146, 276284. frank.koenen@var.fgov.be
Leifer, I., Hoffmann, B., Hoeper, D., Bruun

Rasmussen, T., Blome, S., Strebelow, G.,
Hreth-Bntgen, D., Staubach, C., Beer, M.
(2010), Molecular epidemiology of current
classical swine fever virus isolates of wild
Partners
ORGANISATION
AND CONTACT ADDRESS CONTACT
(deputy coordinator), Lindholm, Tel. +45 35887993
DTU-Vet, se Uttenthal, PhD, Prof., Kalvehave E-mail: ASUT@VET.DTU.DK
Head of NSFL, DTU-Vet, Havnevej 51,
Dept of Virology 4771 Kalvehave, DENMARK
Anses, Anses, Tel. +33 296016290
Marie-Frdrique Le Potier, Dr, PhD, 2731 Avenue du E-mail: marie-frederique.
Head of Gnral Leclerc lepotier@anses.fr,
the Swine Immunology 94701 Maisons Alfort,
and Virology Unit FRANCE
CAO-DVMP, CAO-DVMP, Tel. +36 14330827
Gbor Kulcsr, Budapest E-mail: kulcsar@oai.hu
Dr, DVM, Director Keleti K. u. 24.
1024
HUNGARY
Willie Loeffen, CVI, Tel. +31 320238800
Dr, Project Leader of the Postbus 65, E-mail: willie.loeffen@wur.
Classical Swine Fever 8200 AB Lelystad, nl
Group and Head of the CSF NETHERLANDS
Research Group (CVI)
FDV (subsidiary company of Pfizer Fort Dodge Veterinaria Tel. +34 972293069
Inc), Juan Plana Durn, S.A., E-mail: joan.plana@pfizer.
Senior Technical Director and Senior Carretera Camprodon s/n com
Director Finca La Riba,
of R & D 17813 Vall de Bianya,
Girona,
SPAIN
FLI, FLI Insel Riems Institute of Tel. +49 3835171200
Martin Beer, DVM, Diagnostic Virology, E-mail: Martin.Beer@fli.
PD Dr. med. vet., Sdufer 10, bund.de
Head of the Institute of Diagnostic 17493 Greifswald-Insel
Virology Riems, GERMANY
IZS-UM, IZS-UM, Tel. +39 075343239
Gian Mario De Mia, Via G.Salvemini 1, 06126 E-mail: gm.demia@izsum.it
Dr, Director of NSFL Perugia,
ITALY
IVI, Sensemattstrasse 293, Martin.Hofmann@ivi.admin.
Martin Hofmann, DVM, 3147 Mittelhusern, ch
Dr, Head of the Development SWITZERLAND
Department of IVI
ONCFS, ONCFS, sophie.rossi@oncfs.gouv.fr
Sophie Rossi, PhD, Micropolis, la Brardie,
Unit sanitaire de la faune (Wildlife Belle Aureille 05000 GAP,
Sanitary Unit) FRANCE
SCU, Wangjiang Road 29, gaorong96@gmail.com
Rong Gao, Prof., 610064 Chengdu, CHINA
Key Laboratory for Bio-Resource
and Eco-Environment of Ministry of
Education Life Science College SCU
ORGANISATION
AND CONTACT ADDRESS CONTACT
SVA, Immunobiology and Para- Sandor.Belak@sva.se,
Sndor Belk, Prof., sitology SVA, Ulls vg 2B, Frederik.Widen@sva.se
Frederik Widn, Dr, and Lihong Liu, SE-751 89 Uppsala, and
Dr, R & D, Department of Virology SWEDEN Lihong.Liu@sva.se
TiHo, Institute of Virology, Volker.Moennig@tiho-
Volker Moennig, Prof., Bnteweg 17, hannover.de
Dr. med. vet. 30559 Hannover,
GERMANY
UCM, UCM, jmvizcaino@vet.ucm.es
Jose Manuel Snchez-Vizcano, Prof. Dpto. Sanidad Animal,
Avda. Puerta de Hierro s/n,
28040 Madrid,
SPAIN
UFZ, Helmholtz Centre for Envi- hans.thulke@ufz.de
Hans-Hermann Thulke, Dr ronmental research,
Department of Ecological
Modelling,
Permoserstr. 15, 04318
Leipzig, GERMANY
Spectos, Niels Delater Spectos GmbH, Theater- niels.delater@spectos.com
strae 6, 01067 Dresden,
GERMANY
IZSA_M, Istituto Zooprofilattico p.calistri@izs.it
Paolo Calistri, Epidemiologist Sperimentale dellAbruzzo
e del Molise G. Caporale,
Campo Boario, 64100
Teramo,
ITALY
2.3. African swine fever

[ASFRISK]
Evaluating and controlling

the risk of African swine
fever in the EU
Summary the risk of importation and/or spread of the
Acronym:
African swine fever (ASF) in EU Member disease in EU Member States. Current ASF ASFRISK
States is currently confined to Italy (Sar- serological and molecular diagnosis proce-
Project number:
dinia), it was recently introduced to Cau- dures were evaluated and new antibody and
211691
casian countries and Russia and it is highly nucleic acid-based diagnostic tools were
prevalent in sub-Saharan African countries. developed, including front-line and pen-side EC contribution:
EUR 2 984 712
Serological surveys during outbreaks occur- tests, which will be supplied to diagnostic
ring previously in eastern and southern Afri- facilities in Africa and to animal health lab- Duration:
can regions have shown that although the oratories in the EU and others for the early 42 months
average prevalence of viral infection in the detection of potential ASFV incursions. Addi- Start date:
domestic pigs is high, in many cases serum tionally, studies on ASFV-host interactions 1 April 2008
conversion was not detected by OIE and and the achievement of attenuated recom- Instrument:
commercial ELISA tests. These events sug- binant virus strains opened new insights for Collaborative project
gest that complex epidemiological patterns the characterisation of pig immune mecha-
for ASF are established in those regions in nisms relevant for survival following infec-
that African swine fever virus (ASFV) iso- tion with ASFV and for vaccine development.
lates of higher virulence coexist with others The new strategies and the tools developed
inducing moderate to chronic forms of the within this project have been transferred to
disease, as observed in the past in Europe. ICPC partners and other countries through
local training courses, traineeships and
These epidemiological scenarios, further technology transfer actions.
complicated by the incremented mobility of
people, animals and goods across the globe,
emphasise the serious threat ASF presents Aim
to the growing pig farming sector in Africa The project has been organised into four
and in disease-free EU Member States. Work main tasks aims: (i) to evaluate the cur-
developed allows the characterisation and rent ASF epidemiology in Africa, to develop
a better understanding of the epidemiologi- and validate a generic risk assessment
cal situation of ASFV in Africa, Sardinia and for the introduction of ASF into EU coun-
the Caucasus region as well as the main risk tries and subsequent control strategies;
factors that are contributing to the persis- (ii) to develop and validate new antibody
tence and spread of the disease in those and nucleic acid-based diagnostic tools
territories and provided new tools and strat- for ASF, including front-line and pen-side
egies for the control of ASF and to reduce tests, which will be supplied to diagnostic
facilities in Africa and to animal health lab- support better preventive and control strat-
oratories in the EU for the early detection egies for ASF. For example, the identification
of potential ASFV incursions, in particular of the risk factors and routes of potential
by newly emerging strains; (iii) to charac- introduction and spread of ASF will allow
terise pig immune mechanisms relevant the implementation of risk reduction meas-
for survival and to identify mechanisms of ures in those countries that have the dis-
virus-encoded evasion genes towards the ease or are currently ASF-free.
development of candidate vaccines and
to assess the use of antiviral molecules The current epidemiological situation of
as complementary methods for the con- ASF worldwide has highlighted the need to
trol of the disease; and (iv) to transfer the evaluate the existing serological and molec-
new strategies and the tools developed to ular diagnostic techniques towards the
ICPC partners and to others through train- development of adequate diagnostic tools
ing, workshops and technology transfer of the disease. As no vaccine is available,
activities. the presence of ASFV antibodies is indica-
tive of previous infection and, as antibodies
are produced from the first week of infec-
Results and potential tion and persist for long periods, they are a
applications good marker for the diagnosis of ASF. The
The project has substantially contributed to work developed aimed at confirming the use
increase the knowledge of the epidemiology, of current ELISA diagnostic tests, the devel-
risk factors and best control strategies to opment of new ELISA, the development of
better prevent and control ASFV. There was pen-side tests for anti-ASF antibody detec-
extensive work carried out to characterise tion. Findings obtained have confirmed the
and better understand the epidemiological current serological tests are able to detect
situation of ASFV in Africa, Sardinia and the antibodies induced against ASF in all epide-
Caucasus region as well as the main risk miological situations confirming the robust-
factors that are contributing to the persis- ness of the current antibody detection
tence and spread of the disease in those techniques. However, new ELISA based on
territories. One of the most significant out- ASFV recombinant proteins targeting, inter
puts of this project was the development of alia, the detection of ASFV-anti-IgM anti-
a generic risk assessment framework that is bodies for the early detection of infection
available to evaluate the risk of introduction were developed, standardised and validated
of ASFV into each EU country. This frame- to be further developed as ELISA commer-
work incorporates all concepts and methods cial prototypes. A rapid, one-step immuno-
developed during the project in an easy-to- chromatographic strip (INGEZIM PPA CROM
use Excel file that will be distributed and ) serological pen-side test capable of
may be potentially used for any EU country/ specifically detecting anti-ASF antibodies
policymaker to evaluate the specific risk of in serum specimens has been developed,
ASFV entrance in the country and the most validated and is now commercially avail-
important risk pathways that contribute to able. The immunochromatographic test
that risk. The idea is to provide policymak- provides areliable method for detection of
ers with a useful decision support tool that anti-ASF antibodies with 99 % of sensitiv-
may be easily accessible, implemented and ity and 100% of specificity and confidence
updated. Moreover, participatory methods where laboratory support and skilled per-
have been used to perform risk mapping sonnel are limited. This test will be very
in endemic countries (Africa and Caucasus useful as a simple, robust and cheap tool
region) that will support more cost-effective for rapid in-farm ASF diagnosis in countries
preventive and control strategies in the that are endemically infected. In particu-
affected territories. Results obtained will lar, it will be essential for African countries
where the delay for the shipment of sam- The project aimed also towards the imple-
ples to expert laboratories can be long and mentation of a new strategy for the devel-
the use of more sophisticated tests some- opment of vaccines against ASF. Devel-
times problematic. In parallel with this work, opment of vaccines against ASF using
the question of the sample preservation classical approaches through viral inacti
was addressed during the project. Indeed, vation or viral attenuation has shown to
in tropical countries, a cold chain is some- be unsuccessful since the disease was first
times difficult to maintain during the ship- reported. None of the treatments applied to
ment of the samples. Therefore, the use of produce inactivated immunogens afforded
filter paper for blood collection, drying and levels of protection acceptable for a vac-
storage at high temperature (> 30 C) was cine. The attempts to obtain ASFV attenu-
evaluated. This support was proved to be ated by either serial passage in cell cultures
suitable for virus and nucleic acids preser- or alternate passages in swine and rabbits
vation. It can be also used with the newly have produced immunogens able to protect
developed molecular tests described below. swine against the fully virulent homologous
virus. However, all the protective attenuated
Further to the use of antibody detection ASFV obtained showed a residual level of
tests, the molecular diagnosis of ASF relies virulence unacceptably high to be used as
so far on a limited number of PCR methods. a vaccine. Two main areas were considered
The updating of current molecular diag- in this project as a major strategy for the
nostic techniques has been a main issue. A development of immunogens against ASF:
real-time PCR method using a commercial
Universal Probe Library (UPL-PCR) probe, a the characterisation of viral host inter-
Linear-After-The-Exponential PCR (LATE- actions through the study of the role of
PCR) assay, and a Loop-mediated isother- virus and host genes in infection (modu-
mal amplification (LAMP) system have been lating host defences, immune evasion,
developed and evaluated for their applica- virus productivity and virulence) towards
tion in the molecular diagnosis of ASF. All the obtainment of attenuated virus
assays enable the detection of different strains;
ASFV p72 genotypes tested, showing as well the in vitro and in vivo characterisation
significant levels of sensitivity and specific- of pig immune mechanisms relevant for
ity. The developed molecular methods, and animal survival against ASFV.
specifically the UPL-PCR and the LAMP tech-
niques, have been adapted as commercial This work revealed further insights into
diagnostic kits and are under evaluation for the ASFV protection model selected for
their upcoming launching in the market. this study: two ASFV isolates of different
virulence (ASFV/L60 and ASFV/NHV/P68,
So, a range of valuable molecular and referred to as L60 and NHV respectively), in
serological tools has been produced within which NHV has previously shown to induce
the ASFRISK project, being now offered to pig survival against L60 infection, but with
improve and complement the available a residual virulence that must be eliminated
ASF diagnostic tools, suitable for use in for vaccination purposes.
well-equipped international and national
reference laboratories, in basic regional Several cell models have been optimised
and local laboratories, or even for rapid for their use in growing and titrating many
on-site application. This will assist Afri- ASFV strains (both from the field or labora
can countries and others to improve their tory) and can be used to infect directly with
diagnostic capacities and will contribute to the field isolates in an established cell line
improve knowledge on the epidemiology of to amplify virus samples either for diag-
the disease. nosis or infection studies, and it was the
basis to generate and purify virus dele- A major tool expected to be delivered by the
tion mutants from the NHV virus model by project is the development of an attenu-
homologous recombination. A number of ated model for protective immunity. NHV
new recombinants from different paren- recombinants deleted of specific genes
tal viruses deleted of specific genes were (A238L, A224L and EP153R) were obtained
constructed and used in in vitro studies to in order to reduce the residual virulence of
analyse the role of the virus genes in the the parental NHV isolate and are currently
evasion of the antiviral response. used in an in vivo experiment to determine
the possible protection induced against
Another achievement was the initiation other virulent ASFV isolates. This eventu-
of the cloning of the ASFV genome into a ally should lead to a strategy to identify the
Bacterial Artificial Chromosome (BAC), an best attenuated virus model to construct an
important tool to facilitate the manipula- effective vaccine against ASF.
tion of virus genes and the generation of
recombinant viruses with specific genes As an alternative approach to the vaccine
deleted by a non-conventional approach. development, a number of antiviral drugs
The sequences of many ASFV genes includ- were studied to be used to contain the virus
ing the open reading frames and flanking spreading from the ASFV outbreaks. Sev-
regulatory regions in the NHV and L60 eral antivirals have been demonstrated to
isolates, were obtained during this project be effective in the inhibition of in vitro ASFV
and used for the generation of the ASFV infection: (i) chemicals directed against
deletion mutants. viral DNA polymerase; (ii) siRNA against
different virus components (A151R gene or
Regarding the control of the host antiviral vp72); and (iii) non-conventional antivirals
response, we have confirmed the role of like lauryl gallate, affecting cellular fac-
different ASFV genes involved in immune tors required for the productive infection.
response (A238L, EP153R, K205R, I329L, All of them were found to be selective and
etc.) and in the control of apoptosis and cell non cytotoxic at the concentrations used to
cycle (A224L, EP153R, g5R, etc.). Specifi- inhibit ASFV infection, and several of them
cally, the following have been determined: have been selected to be tested in an in vivo
(i) the immunomodulatory role of A238L experiment to analyse the possible protec-
gene, confirming the inhibition of NF-KB and tion of pigs pre-treated with the antivirals
NFAT transcription factors through a novel after a challenge with virulent ASFV isolates.
mechanism involving protein kinase-C-medi-
ated p300 transactivation; (ii) the effect of ASFRISK was also dedicated to the organ
EP153R gene in the modulation of the SLA-I isation of training and technology transfer
expression, including 3D modelling of the activities coordinated by EU partners in col-
interaction between the viral lectin and the laboration with the African and Asian part-
MHC-I molecules, and the analysis of the ners of the consortium. Under Local train-
critical regions involved in the inhibition; ing/workshops and technology transfer, five
and (iii) the role of K205R, I329L and MGF- training courses on clinical and laboratory
360-18R virus genes in the inhibition of IFN diagnosis and ASF epidemiology-modelling
induction and signalling and in the NF-kB were organised in China, Ivory Coast, South
activation, including the analysis of different Africa, Spain and Uganda. These courses
structural regions of the viral proteins in the were very successful both for training on
modulation of these processes. The modula- relevant topics for the diagnosis, preven-
tion of specific cytokines, chemokines and tion and control measures of ASF in dif-
inflammatory molecules, induced during ferent scenarios and for the reinforcement
virus infection of porcine macrophages, has or establishment of working links between
also been analysed. the EU partners in the project and countries
in other parts of the world (different coun- S., Arias, M., De Mia, G., Genetic character
tries in Africa, Belarus, China, Russia and isation of African swine fever viruses from
Vietnam) for which ASF is a great matter recent and historical outbreaks in Sardinia
of concern. Technology transfer mainly (19782009), 2011, Virus Genes, 42(3):
based on the most recent validated labo- 37787.
ratory diagnostic tests and on epidemio-
logical tools developed within the ASFRISK Hurtado, C., Bustos, M.J., Carrascosa, A.L.
project was included in the different train- (2010), The use of COS-1 cells for studies
ing courses, traineeships and on a ring trial of field and laboratory African swine fever
on the easy and rapid LAMP PCR recently virus samples, Journal of Virological Meth-
performed. ods, 164: 131134.
Further to the above, an interactive CD on Mur, L., Martnez-Lpez, B., Martnez-Avilz,

ASF diagnosis available in English, French, M., Costard, S., Wieland, B., Pfeiffer, D.U.,
Russian and Spanish was produced by the Snchez-Vizcano, J.M., Quantitative Risk
Faculty of Veterinary Medicine of Madrid Assessment for the Introduction of African
(OIE reference laboratory) with the collabo- Swine Fever Virus into the European Union
ration of CISA-INIA (EU reference laboratory by Legal Import of Live Pigs, Transboundary
for ASF). This CD has been used for teach- and Emerging Diseases, 2011, 10 August,
ing purposes during training activities of the doi: 10.1111/j.18651682.2011.01253.x.
project and distributed to several institutions (Epub ahead of print).
worldwide. Individual Long-Term Training
offered one MSC training in South Africa, Ronish, B., Hakhverdyan, M., Sthl, K., Gal-
three PhD trainings in Ivory Coast, Portugal lardo, C., Fernandez-Pinero, J., Belk, S.,
and South Africa and another one long-term LeBlanc, N., Wangh, L. (2011), Design and
training (six months) for a Chinese fellow in verification of a highly reliable Linear-After-
Spain. In parallel, four short-term trainings The-Exponential PCR (LATE-PCR) assay for
were organised for two Chinese and two the detection of African swine fever virus,
Vietnamese fellows in Sweden and Spain. Journal of Virological Methods, 172, (12):
815.
The work achieved under the different tasks
of the ASFRISK project has contributed to Project website
the reinforcement of EU scientific and tech- http://www.asfrisk.eu
nological capacities and to the establish-
ment of links with African, Asian and east- Keywords
ern European countries that will also benefit ASF, ASFV, Africa, epidemiology, risk assess-
from the achievements of the project. ment, molecular diagnostics, antibody
detection, vaccine development, immunity
References/publications Coordinator
Gallardo, C., Anchuelo, R., Pelayo, V., Poude- Prof. Carlos Martins
vigne, F., Leon, T., Nzoussi, J., Bishop, R., Faculdade de Medicina Veterinria,
Prez, C., Soler, A., Nieto, R., Martn, J.H., Universidade Tcnica de Lisboa
Arias, M. (2011), African swine fever Av. da Universidade Tcnica
virus p72 genotype IX, in domestic pigs in 1300477 Lisboa
Congo, Emerging Infectious Diseases, 17(8): PORTUGAL
15561558. cmartins@fmv.utl.pt
Giammarioli, M., Gallardo, C., Oggiano, A.,

Iscaro, C., Nieto, R., Pellegrini, C., Dei Giudici,
Partners
ORGANISATION ADDRESS E-MAIL
Fundao Calouste Rua da Quinta Grande 6 parkhous@igc.gulbenkian.pt
Gulbenkian Oeiras
Instituto Gulbenkian de 2781-901
Cincia PORTUGAL
Universidad Complutense de Veterinary School jmvizcaino@visavet.ucm.es
Madrid Av. da Puerta De Hierro
Animal Health Department 28040 Madrid
SPAIN
Instituto Nacional de Crta de Algete a El Casar, arias@inia.es
Investigacin km 8,100
Agraria y Alimentaria 28130, Valdeolmos
SPAIN
Consejo Superior de Nicolas Cabrera, 1 acarrascosa@cbm.uam.es
Investigaciones Cientficas 28049 Cantoblanco Madrid
Centro de Biologa Molecular SPAIN
Severo Ochoa
Centre de coopration TA A-15/Gt emmanuel.albina@cirad.fr
internationale en recherche 34398 Montpellier
agronomique pour le FRANCE
dveloppement
Campus International de
Baillargue
Istituto Zooprofilattico Via Salvemini 1 gm.demia@izsum.it
Sperimentale dellUmbria e 06126 Perugia
delle Marche ITALY
National Swine Fever
Laboratory
Friedrich-Loeffler-Institut Boddenblick 5a guenther.keil@fli.bund.de
Federal Research Institute for 17493 Greifswald-Insel Riems
Animal Health GERMANY
Royal Veterinary College Hawkshead Lane Pfeiffer@rvc.ac.uk
Veterinary Clinical Sciences North Mimms, AL9 7TA
UNITED KINGDOM
The National Veterinary Ulls Vag 2B sandor.belak@sva.se
Institute SE-751 89 Uppsala
SWEDEN
The Agricultural Research Old Soutpan Road, 1 HeathL@arc.agric.za
Council 0110, Onderstepoort, Pretoria
Transboundary Animal SOUTH AFRICA
Diseases Programme
Laboratoire National dAppui Rue du Jardin Botanique e.couacy-hymann@lanada.ci
au Developpment Agricole BP 206, Bingerville
Laboratoire de Virologie CTE DIVOIRE
Veterinary and Agrochemical Groeselenberg 99 frkoe@var.fgov.be
Research Centre 1180 Brussels
BELGIUM
ORGANISATION ADDRESS E-MAIL

Lanzhou Veterinary Research Xujiaping 1 ttbdcn@public.lz.gs.cn
Institute Lanzhou, Gansu
Chinese Academy of 730046
Agricultural Sciences CHINA
National Institute for Truong Chinh, 86 dzungntd@gmail.com
Veterinary Research Dong Da Hanoi
VIETNAM
Inmunologia Hnos. Garcia Noblejas cvela@ingenasa.es
y Genetica Aplicada S.A. 28037 Madrid
SPAIN
Department of Veterinary Stoney Road, Stormont Gordon.allan@afbini.gov.uk
Sciences Belfast
Queens University of Belfast BT4 3SD
UNITED KINGDOM
2.4. Orbivirus
[BTVAC]
Improved vaccines for

bluetongue disease
Summary of vaccines that will protect against all BTV
Acronym:
Bluetongue is a viral disease of livestock, serotypes. All partners made significant BTVAC
particularly sheep that is endemic in many scientific progress in meeting the aims and
Project number:
tropical and sub-tropical countries and objectives of the project. The effect of the
044211
can result in high morbidity and mortality. inactivated BTV vaccine on pregnant ewes
Outbreaks have been frequent in Southern and lambs has been determined with no EC contribution:
EUR 840 000
Europe since 1998, and in 2006 the virus clinical side effect recorded. Substantial
emerged as far north as France, Belgium, progress has also been made in under- Duration:
Holland and Germany, reaching the UK in standing the immune response to the sin- 45 months
2007. Existing vaccines, based on attenu- gle BTV VLP vaccine in sheep and demon- Start date:
ated strains of the causative virus, are strating that it provides protection against 1 January 2007
protective but cause viraemias and terato- clinical symptoms and viraemia. The BTV Instrument:
logical effects when pregnant animals are VLP vaccines with more than one serotype, Specific Targeted
vaccinated. have also been successful in a clinical trial Research or
with the animals protected from clinical Innovation Project
(STREP)
The primary aim of this project was, there- symptoms and viraemia. This indicates that
fore, to generate completely safe, geneti- there is no interference in the immunologi-
cally inert vaccines for BTV, initially for cal response to the VLPs. Both the BTV VLP
serotypes that are identified in Europe and inactivated BTV vaccines are promising
with a future aim to generate a portfolio candidates. Preliminary results from field
sera highlight the potential use of a NS1 variable protein in the virus. After entry
competitive ELISA developed within this into cells, the outer capsid (VP2, VP5) is
framework and VP7 ELISA as a DIVA test. removed to release a transcriptionally
Real time RT-PCR detections of BTV have active core particle composed of two major
also been developed, all showing highly structural proteins (VP7 and VP3) and the
sensitive detection methods. transcription complex of three enzymatic
proteins (VP1, VP4 and VP6) in addition to
the segmented dsRNA genome.
Problem
Bluetongue virus (BTV) is the cause of blue- Like almost all other RNA viruses, the BTV
tongue (BT) disease, an insect-vectored genome shows a rapid rate of sequence
emerging pathogen of wild ruminants and mutation. In addition, each virus particle
livestock causing disease in sheep, goats, contains 10 different RNA segments and
and cattle with mortality reaching 70% in these segments reassort when two dif-
some breeds of sheep. BTV infection occurs ferent viruses infect the same host. Thus,
throughout many of the temperate and BTV reassorts freely and exchange readily
tropical regions of the world, coincident occurs between different serotypes.
with the distribution of specific species of
Culicoides biting midges that act as biologi- On the basis of serotype-specific virus
cal vectors of the virus. Bluetongue virus is neutralization assays, 24 distinct sero-
the prototype member of the genus Orbivi- types of BTV have been described to
rus, family Reoviridae. date, although the Toggenburg virus iso-
lated from goats in Switzerland, is prob-
ably a 25th serotype. BTV is endemic in
many tropical and sub-tropical countries.
Since 1998 there have been separate and
repeated incursions of bluetongue into
Europe and 6 serotypes (BTV-1, -2, -4,
-8, -9 and -16) have been introduced into
mainland Europe and the Mediterranean
region. 2006 was a landmark year as it
saw the emergence of BTV (serotype 8)
as far north as France, Belgium, Holland
and Germany and subsequently across
the English Channel to the UK in 2007. Cli-
mate change may have contributed to the
emergence of BTV in Europe through the
increased distribution and size of insect
vector populations. It is also evident that
BTV is being transmitted by novel vector
Bluetongue virus has a complex multi- species, which are abundant in central and
layered architecture (Figure 1). Virions northern Europe. Thus, BTV now repre-
contain non-equimolar amounts of 7 pro- sents a considerable threat in all European
teins organised into two capsids. The outer countries including the UK.
capsid, composed of two major structural
proteins (VP2 and VP5), is involved with cell The European incursion of BTV has had a
attachment and virus penetration during considerable negative economic impact,
the initial stages of infection. The cellular partly due to high mortality and associ-
attachment protein, VP2, is also the sero- ated high morbidity but, more importantly,
type determinant of BTV and is the most as a result of the ban of trade between
BTV-infected and non-infected areas. Results

Based on the number of ruminants, the In order to achieve the aim and objectives
annual turnover of an efficient BTV vac- of the project, Merial (Partner 2) devel-
cine is estimated to be as much as 16m oped an inactivated virus-based BT vac-
within Europe. To limit direct losses and in cine, effective against a number of BTV
an effort to minimize the circulation of BTV, strains (BTV-2, -4 and -8), that has been
European countries have vaccinated live- used successfully during the 2006-2008
stock with available vaccines. European outbreaks, although not in all
potential ruminant hosts. As there was no
The threat of BT disease to European agri- data on the safety and efficacy of these
culture has stimulated the development of vaccines in pregnant animals, a clinical
new and safer vaccines against BTV. The trial was undertaken. The data obtained
purpose of this EC funded project, specifi- clearly demonstrated that these killed-
cally, was to test the safety of inactivated virus vaccines are also safe in pregnant
vaccines in pregnant animals and develop ewes, in contrast to live virus vaccines.
new types of sterile (protein based) vac- In parallel, LSHTM (Partner 1) developed
cines to Bluetongue. novel complex protein-based recombi-
nant virus-like particle (VLP) vaccines for
a number of European strains (BTV-1, -2,
Aim -4, -8 & -9). VLPs have been proven to
The aims of this project were to generate substantially improve immunogenicity and
completely safe genetically inert vaccines elicit stronger and longer-lasting immune
for BTV, particularly for European sero- responses (both B- and T-cell). Moreover,
types, either by inactivation of the infec- because the VLPs contain only the pro-
tious virus or by recombinant technology tein, and not the viral genome, there is no
that completely avoids infectious virus. The chance of reversion to virulence, reassort-
specific objectives of the project were: ment or incomplete inactivation. Several
of these VLPs were tested by UCM, ANSES
1. To establish the safety of vaccination and AUTH (Partners 3, 4 & 5 respectively)
with inactivated vaccines in pregnant in three different European breeds of
animals. sheep that are susceptible to BTV. Vacci-
2. Validated the protective efficacy of nation with monovalent, divalent or mul-
VLP vaccine in European sheep, cattle tivalent (VLPs of two or more different
and goat breeds against virulent virus serotypes) VLPs were capable of protect-
challenge. ing sheep successfully from BT disease
3. To characterise the type of immune when challenged with virulent viruses. In
response elicited by vaccination with each case viraemia was completely sup-
inactivated and VLP vaccines. pressed. Moreover, there was no interfer-
4. To determine the breadth of protection ence observed from mixed serotype vac-
afforded by inactivated vaccine and VLP cines, when comparing the monovalent
vaccine in sheep. and polyvalent VLP vaccines. The VLP vac-
5. To determine the degree of cross- cinated sheep showed complete protection
neutralisation afforded by a mixture of against virulent BTV strains. Furthermore,
VLPs to all current European serotypes there was no interference in protection
against homologous and heterologous when sheep were vaccinated with more
serotypes. than one type of VLPs. Thus, demonstra-
6. To develop a test, based on BTV NS1 tion of the effectiveness of VLPs as a
and VP7, capable of distinguishing vaccination strategy in European animals
vaccinated versus naturally infected has been successfully achieved. Since it is
animals. essential to develop a test that is capable
of distinguishing vaccinated and infected 6. Important data on the safety of inacti-

animals (DIVA test), IDVET (Partner6) vated vaccine in pregnant animals.
focused on the development of serologi- 7. A prototype kit for BT capable of dis-
cal diagnostic tests for discriminating vac- tinguishing vaccinated and infected ani-
cinated versus naturally infected animals mals based on NS1.and VP7
as well as developing real time RT-PCR 8. New patent for new technology to gen-
detections of BTV together with VAR (Part- erate BTV VLPs.
ner 7). Within this EC project, LSHTM also 9. VLPs production technology has been
constructed VLPs for the eastern (Greece successfully transferred to a vaccine
isolate; GRE) and western (South African manufacturing company.
isolate; RSA) lineages of BTV-1, BTV-2 10. Further development and validation of
and BTV-4 which were validated in clinical real time RT-PCR detection and quanti-
trials using European breeds of sheep. In fication of BTV.
addition, the degree of cross-neutralisation 11. A strengthening of the ties between
achieved from VLPs vaccines containing the research and commercial partners
mixtures of serotypes was investigated. in the project promoting the transfer of
Tests capable of distinguishing vaccinated technology from a research to a pro-
and infected animals were also developed duction phase.
and supplied to partners for the compara-
tive vaccine trials. In conclusion, the project has been
extremely successful both scientifically
and for practical purposes and substantial
Potential applications achievements towards the objectives of the
The proliferation of BTV severely limits project have been obtained.
sheep production and the live sheep and
cattle export business; countries free from
virus will not accept animals from coun- Keywords
tries where the disease has broken out bluetongue, non-live vaccine, VLPs, DIVA,
or is endemic with substantial economic diagnostics
implications. As a result of the BTVAC pro-
ject, the following Outcomes have been Publications
achieved; 1. Prez de Diego AC, Athmaram TN, Stew-
art M, Rodrguez-Snchez B, Snchez-
1. A better knowledge of the type of Vizcano JM, Noad R, Roy P. 2011. Char-
immunity elicited by the vaccines in the acterization of protection afforded by
study. a bivalent virus-like particle vaccine
2. Production of reagents for the rapid against bluetongue virus serotypes 1
generation of a marketable novel VLP and 4 in sheep. PLoS One. 6(10):e26666
vaccine for all the circulating European 2. Stewart, M., Y. Bhatia, T. N. Athmaran,
serotypes of BTV. R. Noad, C. Gastaldi, E. Dubois, P. Russo,
3. Evidence that single and/or multiple R. Thiery, C. Sailleau, E. Breard, S. Zien-
serotype VLP vaccines are highly effica- tara, and P. Roy. 2010. Validation of a
cious against a virulent virus challenge. novel approach for the rapid production
4. There was no interference in the gener- of immunogenic virus-like particles for
ation of neutralizing antibodies or pro- bluetongue virus. Vaccine 28:3047-54.
tection afforded by a multiple serotype 3. Vandenbussche, F., T. Vanbinst, E. Van-
VLP vaccine. demeulebroucke, N. Goris, C. Sailleau,
5. Demonstration that VLPs afford protec- S. Zientara, and K. De Clercq. 2008.
tive efficacy (no clinical sign or viraemia) Effect of pooling and multiplexing on
in 3 different European sheep breeds. the detection of bluetongue virus RNA
by real-time RT-PCR. J Virol Methods. P. Kerkhofs, N. Goris, and F. Vanden-

152:13-7. bussche. 2008. Transplacental infec-
4. Chatzinasiou, E., C. I. Dovas, M. Papa- tion and apparently immunotolerance
nastassopoulou, M. Georgiadis, V. Psy- induced by a wild-type bluetongue virus
chas, I. Bouzalas, M. Koumbati, G. Kop- serotype 8 natural infection. Trans-
topoulos, and O. Papadopoulos. 2010. bound Emerg Dis 55:352-9.
Assessment of bluetongue viraemia in
sheep by real-time PCR and correlation Coordinator
with viral infectivity. J Virol Methods. Prof. Polly Roy
169:305-15. Professor of Virology
5. De Clercq, K., I. De Leeuw, B. Verhey- London School of Hygiene and Tropical
den, E. Vandemeulebroucke, T. Vanbinst, Medicine
C. Herr, E. Meroc, G. Bertels, N. Steur- Keppel Street
baut, C. Miry, K. De Bleecker, G. Maquet, London WC1E 7HT
J. Bughin, M. Saulmont, M. Lebrun, B. UK
Sustronck, R. De Deken, J. Hooyberghs, Polly.roy@lshtm.ac.uk
P. Houdart, M. Raemaekers, K. Mintiens,
Partners
ORGANISATION CONTACT E-MAIL
MERIAL SAS Dr Pascal Hudelet Pascal.hudelet@merial.com
29, avenue Tony Garnier
69007 Lyon
FRANCE
Universidad Complutense de Prof Jose Manuel jmvizcaino@vet.ucm.es
Madrid Sanchez-Vizcaino
Ciudad Universitaria
28040 Madrid
SPAIN
ANSES Dr Stephen Zientara szientara@vet-alfort.fr
27-31 Avenue du Gnral Leclerc
94701 Maisons-Alfort Cedex
FRANCE
Aristotle University of Dr Maria mpapanas@vet.auth.gr
Thessaloniki Papanastasopoulos
541 24,
GREECE
IDVET Dr Phillipe Pourquier Phillipe.pourquier@id-vet.com
Montpelier
FRANCE
Veterinary and Agrochemical Dr Kris de Clerc krdec@var.fgov.be
Research Centre
Ukkel Campus
Brussels
BELGIUM
[Medreonet]
Surveillance network of
reoviruses, bluetongue and
African horse sickness,
in the Mediterranean
basin and Europe
Summary to Europe since only serotype-specific vac-
Acronym:
Medreonet
Bluetongue virus (BTV) and African horse cines are available (http://www.reoviridae.
sickness virus (AHSV) are reoviruses trans- org/dsRNA_virus_proteins/ReoID). A new
Project number:
mitted by vectors species belonging to the orbivirus named BTV-25 (Hofman et al.,
044285
Culicoides genus that affect respectively 2009; Hofman et al., 2010; Chaignat et
EC contribution: ruminants and Equidae. al., 2009) was described in Switzerland
EUR 460 000
in 2008. The importance of implement-
Duration: BT disease has occurred sporadically in the ing surveillance networks has to be noted
42 months Mediterranean region in the past, involving in the way it helps the countries to detect
Start date: relatively short-lived epizootics. Since 1998, bluetongue occurrence even if the clinical
1 January 2007 large BT outbreaks affected different coun- signs were moderate or even absent for the
Instrument: tries around the Mediterranean. The virus newly detected serotypes.
coordination action has extended further north than ever. This
geographical expansion is mainly due to the AHSV outbreaks have occurred in south-
northern extension of the main afro-trop- ern Europe in the past, especially in Spain
ical BT vector C. imicola. During summer from 1987 to 1991. It causes one of the
2006, BT outbreaks due to serotype 8 were most severe diseases in horses. It is closely
recorded in Belgium, Germany, France and related to BTV and is transmitted by the
the Netherlands with European Culicoides same Culicoides vectors; hence regions at
species probably involved in this emer- risk of BTV can be regarded as at risk of
gence. This episode highlighted the poten- AHSV. Recent outbreaks of AHSV serotype
tial of BT to further establish in Europe and 2 occurred in Nigeria and Senegal in 2007
presents a major risk to the livestock indus- and, more recently, in 2010 in Ghana.
try. Two new serotypes appeared in north- Apart from BT and AHS, another Culi-
ern Europe: BTV-6 and BTV-11 in Germany coides-borne virus, Epizootic hemorrhagic
and the Netherlands in November 2008, disease virus (EHDV) has been detected
and in Belgium in early 2009 respectively. in 2004 in Morocco, in 2006 in Israel and
Several other serotypes coming from east- the Maghreb northern countries (Algeria,
ern Europe (mainly Israel) such as serotypes Morocco and Tunisia) and on La Runion
5, 15 (outbreaks in 2009) and serotype 24 Island in 2009 and 2010. Dairy cattle (Hol-
(outbreaks in 2009 and 2010) are a threat stein and Brown Swiss) were suspected to
have EHD on the basis of clinical investi- Mediterranean basin and in the north of
gation between early July and the last Europe.
week of August 2007 in several cities in
the western part of Turkey, the cases were The aim of this project is to promote the
confirmed (Temizel et al., 2009). The coor- coordination of efforts directed at improv-
dination action will gather and share infor- ing knowledge relevant to the control of
mation on BT, AHS and EHD to: (i) promote bluetongue (BT), African horse sickness
regional studies on the risks of introduc- (AHS) and Epizootic Hemorrhagic disease
tion of new strains and spread with inclu- (EHD). It is intended to strengthen the
sion of neighbouring areas (North Africa, surveillance of these three Culicoides-
Turkey) as an early warning; (ii) survey the borne diseases by providing a frame-
expansion of C. imicola in new northern work forinteractions between research
territories taking into account the poten- institutions and national veterinary ser-
tial novel vectors group in Europe; and (iii) vices. Theproject will focus on the coor-
improve information technology for stor- dination of research, disease surveillance
age, communication and sharing of vector at the Mediterranean level, risk analysis
and sentinel surveillance and vaccination through network resources, information
data. The consortium is bringing together exchange on technical issues and dedi-
national and international reference labo- cated studies.
ratories working on vectors, detection of
infection, and surveillance and risk assess- In order to answer the objectives of the
ment around the Mediterranean. call, six work packages have been identified
divided into vertical activities dealing with
either tools or data needed for surveillance,
Problem and horizontal or transversal activities
A total of 21 partners with two giving us a dealing with more integrative activities.
hard time by not returning the administra-
tive documents and one which did not pro- The vertical activities are:
vide any input to the project.
regional surveillance of virus activity
and vaccination WP1;
Aim regional surveillance of vectors WP2;
The strategic objectives of Medreonet are: molecular Epidemiology WP3.
to further expand our knowledge of the The transversal activities are:

epidemiology of bluetongue, African
horse sickness and Epizootic Haemor- databases, web design and GIS WP4;
rhagic Disease in the Mediterranean risk assessment WP5;
basin and Europe; meetings WP6.
to apply this knowledge to optimise the
surveillance of these Culicoides-borne
diseases. Results
1. Scientific
The research objectives of Medreonet are: Improvement of the surveillance net-
work for BT, AHS and EHD in the Euro-
to further study the ecology of Culicoides Mediterranean region
vectors and their expansion; Data on vector distribution
to further study the behaviour of blue- Data on viral strain distribution
tongue and Epizootic Haemorrhagic Data on potential threatening viral strain
Disease after their emergence in the around the region
2. Technical and policy support For African horse sickness and Epizootic
Recommendations on implementation of Hemorrhagic Disease, the potential impacts
national surveillance systems are:
Better documentation of disease status
Harmonisation of diagnostic tests and enhance the knowledge on AHS and EHD
case definition in zones at risk;
Development and maintenance of Euro- disseminate information about AHS and
Mediterranean database on viral strain EHD in the Euro-Mediterranean countries
and disease situation and Europe;
increase the rapidity of early warning
The potential impacts are to strengthen when a new strain is introduced in one
and coordinate existing initiatives for col- area;
laborative actions involving national ref- assess the risk from source of AHS and
erence laboratories, OIE/EU reference EHD virus from areas surrounding the
laboratories and other international stake- Mediterranean.
holders involved in BT or AHS research
and to initiate new collaborative action on Potential applications
research activities and needs, regional sur- The potential applications of the different
veillance and risk management/research, deliverables that have been achieved during
diagnostic harmonisation and laboratory the project are described by WP:
preparedness; to establish a website act-
ing as a front-end for Euro-Mediterranean WP1: Regional surveillance of virus
situation towards BT, AHS and EHD and activity
a European research group on BT, AHS The general objectives of this WP1 con-
and EHD threats; to provide a manage- sisted in the strengthening and the har-
ment structure to improve the outputs monisation of surveillance of BTV/AHSV/
from collaborative actions and to involve EHDV infection in Europe. The specific
stakeholders in the scientific and technical objectives are the evaluation and the har-
developments. monisation of surveillance protocols and
description of available tools for surveys
More specifically, for bluetongue, the and viral. This general objective was pur-
potential impacts are: sued by evaluating the following three
different components: (i)surveillance pro-
facilitate decision-making based on tocols; (ii) available tools for surveys and
previous experience in Mediterranean strain identification and (iii)vaccination
countries and Europe; strategies and helping all the partners to
enhance the knowledge on BT in zones have a European point of view of the vac-
at risk; cination programmes, case definition and
disseminate information about AHS and laboratory diagnosis of each of the partici-
EHD in the Euro-Mediterranean countries pating countries.
and Europe;
increase the rapidity of early warning A questionnaire was sent out to all the
when a new strain is introduced in one participants to address several ques-
area; tions about BT, AHSV in 2007 in terms
spread knowledge and diagnostic tech- of case definitions, laboratory diagnosis
niques relevant to strain threatening the and surveillance of these diseases in each
Mediterranean basin and Europe; country.
better advice on the use of vaccination
against BT and the choice of vaccine At the end of 2007, in order to avoid dupli-
serotype to be included. cations, it was decided to take advantage
of the EUBTNet system and obtain the development of standardised proto

from EUBTNet the surveillance data cols for detection and estimation of
required bythe WP1 with the following abundance of Culicoides;
justifications: the development of a reference collec-
tion of C. imicola specimens for DNA
data in the EUBTNet system belong to analysis.
the national competent authorities (i.e.
the involved ministries of the Member WP3: Molecular epidemiology
States); The general objectives of this WP3 con-
it would not have been difficult for the sisted in the distribution, origins andmove-
institutions involved in the project to be ment of the different BTV and EHDV strains,
granted the permission to use aggre- by characterisation of well documented iso-
gated data for the aims and scopes of lates of different serotypes.
the project;
data in the EUBTNet system is the offi- Expanding the existing sequence data-
cial information available from the bases for genome segments 2, 6 and
Member States, therefore the use of others of BTV and related orbiviruses,
EUBTNet data guarantees the con- including high quality data for repre-
sistency with the official situation of sentative and well documented isolates
member countries; of different virus serotypes from differ-
since the Medreonet project requires ent geographical locations is one of the
only aggregate data on the basis of application of this project and is very use-
administrative units, the proper man- ful in terms of awareness when new out-
agement of sensible data can be easily breaks occur in orbiviruses free countries.
assured. The characterisation of novel and exist-
ing BTV/EHDV strains from Europe and
WP2: Regional surveillance of the Mediterranean basin is helping in
vectors the determination of their geographical
The general objectives of this WP2 consisted distribution, movement, potential for
in the strengthening of Culicoides entomo- reassortment and their original sources
logical surveillance in Europe and neighbour- (topotypes).
ing countries. The specific objectives are the The identification of diagnostic tools such
evaluation and the harmonisation of: as molecular probes and primers for the
rapid/early identification of new strains-
(i) surveillance protocols; and serotypes is also one of the major appli-
(ii) available tools for trapping, identifica- cations for all of the participants.
tion of vectors and modelling of vector
habitat. WP4: Database, web design and GIS
The general objective of this WP4 was to
The various deliverables that have been develop a Culicoides-borne virus network
achieved could all be considered as through a website which will provide a plat-
potential applications, particularly: form of Internet-based tools to be used by
partners and other interested parties, to
the recommendation of the optimal trap enable:
design for sampling Culicoides;
the development of a guide for the (i) effective information exchange between
identification of Culicoides vectors and partners on technical and adminis-
potential vectors; trative CA-related bluetongue AHS
t he recommendat ion of opt imal and EHD activities (restricted-access
surveillance protocols for Culicoides; website);
(ii) information related to the CA to be analyses were performed and aimed at the
imparted to the wider scientific com- following potential applications.
munity and to other stakeholders with
an interest in the control of bluetongue Transmission pathways from currently
and AHS (open-access website); affected areas have been developed
(iii) a forum for discussions on the vertical using international standards for risk
work packages: assessments. The pathways cover both
vector-related transmission as well as
to develop a web-enabled geographic spread related to trade.
information system (GIS) to present A review has been conducted with
epidemiological data and to allow a respect to published models that can
rapid spread of information related to be used for the assessment of infection
the diseases; probabilities of specified regions with
to provide a real-time interactive a methodological guideline to be used
mapping system of the main epide- in order to classify specific regions or
miological aspects to facilitate the Member States with respect to BT risk.
decision-making process and man- Validation of the risk assessment using
agement of control activities at cen- case studies in selected countries (Alge-
tral and local level. ria, Bulgaria, Tunisia and Turkey) with the
collection of samples to identify exotic
The online procedure is an application strains of BT/AHS/EHD and trapping
developed through an ASP page providing graphically presented as maps.
a form that participants can fill in when Based on the epidemiology of BT/AHS/
needed. The online updating system has EHD, risk-based surveillance approaches
been implemented in all the web-GIS ser- have been developed. Risk-based sur-
vices developed: BT outbreaks, BT entomo- veillance is defined as a surveillance
logical surveillance, AHS outbreaks, EHD programme that includes risk factors
outbreaks. for increased probability of infection in
specified populations and/or regions as
Through a username and password, users well as the outcome of risk assessments
can be identified and authorised and they conducted. Thus, the results of the risk
can insert all the information related to assessments will be used as input for
their own country. At the moment, data the planning of surveillance. It is antici-
on presence/absence of the disease can pated that sampling intensity, target
be added, but any additional information population and sampling interval in sur-
required by the participants can be eas- veillance programmes are dependent on
ily added in the form and the database the risk category of the Member State
including disease distribution, viral and or region. Different risk analysis mod-
serological activity, entomological activity, els have been built, to determine the
control measures. risk of overwintering and introduction
of bluetongue virus. The results of the
WP5: Risk assessment models determine the factors that have
The general objective of this WP5 was to more importance in the maintenance/
standardise methods for the geograph introduction of the virus and, as a con-
ical assessment of the risk of BT/AHS/EHD sequence, the sub-populations where
spread in the Mediterranean basin and surveillance can be more sensitive in
Europe. detecting the infection: quantitative
assessment of the probability of blue-
In order to obtain a concept for geographi- tongue virus overwintering by horizontal
cal risk assessment of BT/AHS/EHD, several transmission, quantitative assessment
of the probability of bluetongue virus C., Hoffmann B., Eschbaumer, M., Oura,
transmission by bovine semen and C.A., Potgieter, A.C., Nomikou, K., Mertens,
effectiveness of preventive measures, P.P., Full genome characterisation of Blue-
quantitative assessment of the proba- tongue virus serotype 6 from the Nether-
bility of bluetongue by Culicoides intro- lands 2008 and comparison to other field
duced via transport and trade networks. and vaccine strains, PLoS One, 2010, 5(4):
A paper was recently published by Napp. e10323.
et al., 2010, on the quantitative assess-
ment of the probability of bluetongue Napp, S., Gubbins, S., Calistri, P., Allepuz, A.,
virus transmission by bovine semen and Alba, A., Garca-Bocanegra, I., Giovannini, A.,
effectiveness of preventive measures in Casal, J., Quantitative assessment of the
theriogenology. probability of Bluetongue virus overwinter-
ing by horizontal transmission in vectors,
WP6: Meetings and dissemination ruminants or in both: application to Ger-
One tool for the evaluation of the CA many 200607, Veterinary Research, 2011,
is the dissemination of knowledge. In 42(1): 4.
this coordination action, major scien-
tists and laboratories in bluetongue and Venail, R., Mathieu, B., Setier-Rio, M.L.,
AHS, representatives of private compa- Borba, C., Alexandre, M., Viudes, G., Gar-
nies and international organisations as ros, C., Allene, X., Carpenter, S., Baldet, T.,
well as political decision-makers were Balenghien, T., Laboratory and field-based
brought together within each work pack- tests of deltamethrin insecticides against
age. Manyof the participants and project adult Culicoides biting midges, Journal of
associates are also members of various Medical Entomology, 2011, 48(2): 3517.
national and international societies for
virology, entomology and epidemiology. Project website
These organisations were used as dis- http://medreonet.cirad.fr/
semination platforms. Dissemination and
exploitation of the results was therefore Keywords
guaranteed. orbiviruses, Bluetongue (BT), African horse
sickness (AHS), Epizootic Hemorrhagic Dis-
ease Virus (EHDV), diagnostic, surveillance,
References/publications risk assessment
Baylis, M., Parkin, H., Kreppel, K., Carpenter,
S., Mellor, P.S., McIntyre, K.M., Evaluation of Coordinator
housing as a means to protect cattle from CIRAD
Culicoides biting midges, the vectors of Campus International de baillaruguet
Bluetongue virus, Medical and Veterinary TA A 15/G
Entomology, 2010, 24(1): 3845. 34398 Montpellier Cedex 05
FRANCE
Ctre-Sossah, C., Madani, H., Nomikou, K., catherine.cetre-sossah@cirad.fr
Sailleau, C., Sadaoui, H., Maan, S., Maan, N.,
Zientara, S., Mertens, P., Albina, E., Molec-
ular epidemiology of Bluetongue virus
serotype 1 isolated in 2006 from Algeria,
Research in Veterinary Science, November
2010.
Maan, S., Maan, N.S., van Rijn, P.A., van Gen-

nip, R.G., Sanders, A., Wright, I.M., Batten,
Partners
PARTNER COUNTRY CONTACT E-MAIL
AFSSA/ANSES FRANCE Stephan Zientara s.zientara@AFSSA.FR
IZS ITALY Annamaria Goffredo m.goffredo@izs.it
Liverpool UNITED Matthew Baylis Matthew.Baylis@liverpool.ac.uk
University KINGDOM
IAH UNITED Peter Mertens peter.mertens@bbsrc.ac.uk
KINGDOM
SFVO SWITZERLAND Heinzpeter heinzpeter.schwermer@bvet.admin.ch
Schwermer
INMV ALGERIA Hafsa Madani hafsasfr@yahoo.fr
IRVT TUNISIA Soufien Sghaier sghaiersoufien@yahoo.fr
LNEZ MOROCCO Youssef Lhor youssef_lhor@yahoo.fr
CVCRI TURKEY Arife Erturk arifeerturk@hotmail.com
NDRVMI BULGARIA Georgi Georgiev georgivet2@yahoo.com
UIB SPAIN Miguel Miranda ma.miranda@uib.es
CRESA SPAIN Mariana Domingo mariano.domingo@cresa.uab.es
UCM SPAIN Jos Manuel jmvizcaino@vet.ucm.es
Sanchez-Vizcaino
CAVIIIPD GREECE Mickael Patakakis mjpatakakis@gmail.com
UTL PORTUGAL Isabel Fonseca ifonseca@fmv.utl.pt
ARC OVI SOUTH AFRICA Gert Venter VenterG@arc.agric.za
FLI GERMANY Martin Beer martin.beer@fli.bund.de
CODA-CERVA BELGIUM Kris De Clercq kris.declercq@var.fgov.be
CIDC/CVI NETHERLANDS Piet Van Rijn Piet.vanrijn@wur.nl
DFVF DENMARK Anette Botner aneb@vet.dtu.dk
[ORBIVAC]
Development of vaccines
for BTV, EHDV and AHSV
Summary serotypes of each virus as possible along- Acronym:
Orbivirus diseases, particularly bluetongue side ahigh throughput DIVA assay (e.g. an ORBIVAC
(BT), African horse sickness (AHS) and Epi- ELISA). This proposal will use a coordinated
Project number:
zootic Hemorrhagic Disease (EHD) are ser multi-partner approach to address these 245266
ious current (BT) and potential future (AHS, issues, to develop new experimental proto-
EC contribution:
EHD) challenges facing European agricul- type vaccines and diagnostic approaches.
EUR 2 999 729
ture. Bluetongue virus (BTV) is the cause
of BT disease, an insect-vectored emerging Duration:
36 months
pathogen of wild ruminants and livestock Problem
causing disease in sheep, goats, and cat- Since 1998, there have been more than 12 Start date:
tle. BTV infection occurs throughout many separate introductions of BTV into Europe, 1 February 2010
of the temperate and tropical regions of involving at least 10 different virus strains Instrument:
the world, coincident with the distribu- belonging to eight different serotypes collaborative project
tion of specific species of Culicoides biting (types 1, 2, 4, 6, 8, 9, 16 and a new sero-
midges that act as biological vectors of the type, type-25). These events have resulted
virus. Since AHSV and EHDV are genetically in the deaths of more than twomillionani-
closely related to, and are transmitted by mals and have caused substantial eco-
the same insect vectors as BTV, there is a nomic losses to the agricultural economies
clear risk of the potential introduction of of Europe. The outbreak caused by BTV-8,
these other orbiviruses, and indeed other which started in the Netherlands and Bel-
arboviruses, into Europe. gium in 2006, is, by itself, the largest sin-
gle outbreak of bluetongue ever recorded.
Although there are effective inactivated New introductions of the virus into Europe,
vaccines for some of the individual BTV which have been linked to climate change,
serotypes, which are currently used in have occurred almost every year since
Europe, these are not currently available 1998, with the identification of four new
for all serotypes. In addition, no effective virus strains in 2008 alone. New Culicoides
inactivated vaccines are currently licensed species responsible for virus spread have
and available for use in Europe, for either also been identified in central and northern
AHSV or EHDV. Another issue with the cur- Europe, confirming that the whole of the EU
rent vaccines is that they generate an is now at high risk from incursion of these
antibody response to all of the viral pro- diseases.
teins, making assays that can differentiate
infected from vaccinated animals (DIVA) Since AHSV and EHDV are genetically
difficult or impossible to develop. closely related to, and are transmitted by
the same insect vectors as BTV, there is a
Therefore, the major outstanding challenge clear risk of the potential introduction of
of orbivirus vaccine research is to develop these other orbiviruses and other related
vaccines that can afford a broad protec- viruses into Europe. The recent detection of
tive immune response against as many BTV-9 in North Africa, EHDV in Turkey, and
two different strains of AHSV in West Africa, efforts in South Africa and the United
provide further indications of increased States to develop improved vaccines for
risk from these diseases. The continued these diseases where these viruses are also
appearance of new BTV strains in south- a threat. The consortium includes a number
ern, central and northern Europe raises the of industrial partners (Merial, Pfizer, Delta-
question what virus and what serotype will mune, Boehringer) who are already active
arrivenext. in vaccine manufacture for these and other
veterinary diseases, in order to ensure that
Two types of orbivirus vaccine are currently the findings of the research are transferred
commercially available, based on either as soon as possible into commercial vac-
attenuation or inactivation of the live virus. cines for European livestock. The project
Each is effective at controlling disease but also includes two SMEs (IDVET and INGE-
there are concerns over incomplete attenu- NASA), who will be specifically involved in
ation of the live vaccines in the field and the development of DIVA compatible diag-
their ability to readily reassort with field nostic tests.
strains. Both vaccines only confer serotype-
specific protection and the inactivated
vaccines require two doses for effective Expected results
protection of sheep and cattle. Moreover, Based on the latest developments in vac-
both types of vaccine generate an antibody cine and orbivirus research, there are
response to all of the viral proteins, mak- anumber of exciting approaches that offer
ing assays that can differentiate infected the potential to produce effective multiva-
from vaccinated animals) (DIVA) difficult to lent vaccines to orbivirus diseases. In terms
develop. of eliciting immune responses, two com-
plementary approaches will be followed.
One is based on proven observations that
Aim attenuated virus, VLPs and the VP2 pro-
The overall aims of the project are three- tein of BTV and AHSV, are protective. The
fold. The first aim is to develop multivalent second strategy will attempt to map cross
vaccines using different approaches for neutralising domains within BTV and AHSV
orbiviruses responsible for livestock dis- epitopes, and then use a single immunogen
eases, in particular, BTV (25 Serotypes), to elicit cross neutralising protection.
AHSV (9 Serotypes) and EHDV (7 Sero-
types). The second is to understand the One of the consortium partners (Part-
best vaccination strategy to elicit multi- ner1 LSHTM) is a leader in orbivirus
serotype protection for these viruses in reverse genetics and will specifically gen-
livestock and analyse immune responses erate a new class of disabled single cycle
for each of the novel vaccines developed for (DISC) vaccine for BTV that promises
breadth of protection against multiple sero- improved immunogenicity over non-repli-
types. Finally, the project aims to develop cating vaccines and avoids the problems
DIVA compatible diagnostics that will work associated with replicating attenuated vac-
with the new vaccines developed in order cines. A complementary, protein-based,
to differentiate between vaccinated and AHSV multivalent subunit vaccine will also
infected animals. be produced. Partner 1 will also develop
VLP vaccines for EHDV, which have not
The project will use a coordinated multi- previously been reported for this orbivirus.
partner approach to achieve these aims. It Other partners have specific expertise in
builds on specific expertise and reagents the use of canine adenovirus, canarypoxvi-
that are only available within the consor- ruses and capripoxviruses as delivery sys-
tium and links out to other international tems for orbivirus antigens and will test the
use of these systems to deliver optimised proteins as a basis for DIVA diagnostics will
multivalent immunogens. Another partner, also be investigated, as well as a quantita-
in collaboration with one of the industrial tive real-time PCR assay for EHDV and new
partners, will use Parapox as an expres- serological DIVA tests for AHSV.
sion system for the serotype determining
VP2 protein of AHSV. Each of the vaccine
systems that will be tested in the project Potential applications
offers its own advantages either in terms of New generation multivalent vaccines and
ease of transfer to manufacturing, safety accompanying tests that are compatible
or degree of long-term immune response with DIVA will be developed. None of the
elicited. The project will compare the new current commercial vaccine approaches,
and established systems for the ability to inactivated and attenuated virus vaccines,
stimulate a multivalent immune protection are compliant with DIVA principles, and
against multiple virus serotypes. therefore it is necessary for a new genera-
tion of vaccines that overcomes this limita-
The project will incorporate new reagents tion to be developed. The primary expected
and methods for rapid diagnosis and typ- outcome is that stable, multivalent, new
ing of orbivirus outbreaks and to distinguish generation, DIVA-compliant vaccines will be
infected and vaccinated animals. produced.
Current typing of orbivirus diseases is The project will produce and test new
based on RT-PCR approaches. The consor- prototype vaccines to BTV (DISC and mul-
tium plans to develop real-time RT-PCR tiepitope-conserved antigen types), AHSV
assays for all 25 BTV serotypes as well as (multi-epitope-virus vectored, and pro-
microarrays to provide higher throughput tein based multivalent subunit) and EHDV
and improved sensitivity for the detection (virus-like particles). All of the new vac-
and rapid typing of diagnostic BTV sam- cines are multivalent and designed to elicit
ples. The new molecular (array) assays will cross protection against multiple strains of
be independently validated against current virus. Major research findings of the pro-
RT-PCR based systems and reference sam- ject are anticipated to be the degree of
ples from previously identified BTV strains cross protection that is afforded by com-
for speed and accuracy. bined vaccines and by new immunogens
where multiple epitopes from different
The new vaccine approaches developed serotypes have been engineered into the
during the project should all be suitable for same immunogen.
serological tests that differentiate infected
from vaccinated animals (DIVA). The con- In parallel to each of the new vaccines,
sortium will develop serological tests that DIVA-compatible diagnostic reagents have
allow DIVA to be completed for vaccinated been developed which will allow routine
animals. The consortium will also develop testing of vaccinated and imported ani-
ELISA-based tests that allow serotype mals. These have specifically been designed
determination and differentiation between to be compatible with the new vaccines
BTV and EHDV. Sero-group-specific ELISA (above). The project will also address the
tests that can be used to distinguish ani- outstanding issues in the diagnosis of
mals vaccinated against one serotype but orbivirus diseases by developing microarray
exposed to a second serotype of BTV as and improved real-time PCR reagents for
part of a DIVA strategy will also be devel- serotyping of BTV and by producing a new
oped. These tests will be directed at sero- group specific ELISA test that for the first
types currently circulating in Europe. The time provides an immunological test for
possibility of using virus non-structural distinguishing EHDV andBTV.
The consortium comprises several indus- Project website

trial partners, which include the major http://www.orbivac.eu
industrial companies currently manufactur-
ing vaccines for BTV in Europe. Inclusion of Coordinator
these partners in the same consortium will Prof. Polly Roy
facilitate commercialisation of any of the Professor of Virology
new vaccine approaches developed by the London School of Hygiene and Tropical
consortium. The consortium also includes Medicine
active participation from third countries Keppel Street
(South Africa and the United States), with London WC1E 7HT
partners who bring specific expertise to the UNITED KINGDOM
consortium and enhance its scientific and Polly.roy@lshtm.ac.uk
technical excellence.
Partners
ANSES Dr Stephen Zientara szientara@vet-alfort.fr
2731 Avenue du Gnral Leclerc
94701 Maisons-Alfort Cedex
FRANCE
Universidad Complutense de Dr Jose Manuel jmvizcaino@vet.ucm.es
Madrid Snchez-Vizcano
Ciudad Universitaria
28040 Madrid
SPAIN
Groupe Virologie Dr Catherine catherine.cetre-sossah@cirad.fr
CIRAD Ctre-Sossah
Dpartement Systmes Biologiques
FRANCE
MERIAL SAS Dr Pascal Hudelet Pascal.hudelet@merial.com
29 Avenue Tony
Garnier
69007 Lyon
FRANCE
Group Director Business Devel- Mr Peter Jeffries peter.jeffries@pfizer.com
opment and Global Alliances
2325 avenue du Dr Lannelongue
75668 Paris Cedex 14
FRANCE
Intervet International BV Dr Danny Goovaerts danny.goovaerts@sp.intervet.com
Wim de Krverstraat 35
P.O. Box 31
5830 AA Boxmeer
NETHERLANDS
Boehringer Ingelheim Animal Dr Randolph Seidler randolph.seidler@boehringer-
Health GmbH ingelheim.com
Binger Strasse 173
55216 Ingelheim am Rhein
GERMANY

Friedrich-Loeffler-Institut Dr Thomas C. Thomas.mettenleiter@fli.bund.de
Sdufer 10 Mettenleiter
17493 Greifswald-Insel Riems
GERMANY
Central Veterinary Institute of Dr Piet A. van Rijn Piet.vanrijn@wur.nl
Wageningen UR (CVI)
Department of Virology
P.O. Box 65, 8200 AB Lelystad
NETHERLANDS
Deltamune: Roodeplaat Dr Baltus Erasmus baltus@deltamune.co.za
P.O. Box 14167
0140 Lyttelton
SOUTH AFRICA
Kansas State University Dr Juergen A. Richt jricht@vet.k-state.edu
College of Veterinary Medicine
Diagnostic Medicine/Pathobiology
K224B Mosier Hall
Manhattan, KS 66506-5601
UNITED STATES
Institute for Animal Health Dr Peter Mertens peter.mertens@bbsrc.ac.uk
Pirbright Laboratory
Ash Road
Pirbright
Surrey
GU24 0NF
UNITED KINGDOM
Veterinary and Agrochemical Dr Kris de Clerc krdec@var.fgov.be
Research Centre
Ukkel Campus
1180 Brussels
BELGIUM
IDVET Dr Phillipe Pourquier Phillipe.pourquier@id-vet.com
Montpellier
FRANCE
Ingenasa Hnos. Dr Carmen Vela cvela@ingenasa.es
Garcia Noblejas
28037 Madrid
SPAIN
95
CHAPTER 3.
Endemic
diseases
The economic impact and

research synergy from
EU-funded projects
The context disease in the United Kingdom in 2001
In exploring the economic impact from EU- is estimated to have cost EUR 3.5 billion,
funded research projects, it is necessary including costs associated with the culling
to place the research in the context of the of 0.76 million cattle, 4.9 million sheep and
cost of animal diseases, which can have 0.43 million pigs, without mentioning other
devastating socioeconomic effects both species. However, the total cost to the rural
in developed and developing countries. economy may have been in the region of
Infectious diseases directly affect livestock EUR 8 billion as a result of disruption in
production, food security and food safety, trade and impacts on industries such as
trade, rural development, environment, live- tourism. BSE is estimated to have cost the
lihood of farmers and in some cases are of United Kingdom EUR 8.5 billion between
public health concern. Infectious diseases 1996 and 2010. Bluetongue in the Neth-
also have major consequences on animal erlands in 200607 was estimated to
welfare. have cost EUR 155 million. In the case of
zoonotic conditions such as avian influenza
The most obvious costs are those asso- or West Nile fever, the costs would also
ciated with epidemic disease outbreaks. include those associated with human mor-
The outbreak of foot-and-mouth (FMD) bidity andmortality. Table 1 summarises
Table 1: Cost of epidemic disease outbreaks
(million EUR)
FMD CSF FMD FMD FMD

TAIWAN NETHERLANDS KOREA JAPAN URUGUAY
1997 1997/98 2000 2000 2000/01
Government costs
Compensation 141 887 283 0.5
Control 50 103 50 11 15
Private costs
Agricultural sector 1 652 318
Related industries 2 409 447 45
Other 712
Total 4 964 1 755 333 11.5 60
Source: FAO/OECD: FMD = foot-and-mouth disease; CSF = classical swine fever
C H A P T E R 3 . E N D E M I C D I S E A S E S 97
the cost of a range of epidemic disease Human capacity, future

outbreaks. planning, continuity and impact
Considering the research work recently
Costs that are not so obvious relate to completed or ongoing, the theme of devel-
endemic diseases. Costs associated with oping human capacity, planning for the
liver fluke amount to EUR 2.5 billion glob- future, continuity of the research effort
ally per year and the disease is spread- and focusing on the diseases with greatest
ing, with increasing costs, due to climate impact while reacting to evolving situations
change. It is also estimated that mastitis is obvious.
costs the European dairy industry in the
region of EUR 3.5 billion per year or EUR The PARASOL project ensured the genera-
150 per year for each cow. tion of a cohort of new specialists in the
area of internal parasite control. Previously,
Improved awareness of, preparedness for, research in this area had become less popu-
and response to outbreaks are needed for lar because of the effectiveness of the then
the effective management of the threats of available control methods and many new
animal diseases and depend on sound sci- areas of research, such as genomics, had
ence. Whilst we have good tools to control opened up. However, with the development
many diseases, the situation is constantly of resistance to anthelmintics, it became
evolving. The objective of the research clear that there was need for a rethink of
is to provide tools for control where none control methods and to stimulate research
exist today or to develop better tools where in this area to ensure that the expertise was
this is necessary. The economic impact of not eventually lost. The DELIVER, PARASOL
research comes in the form of reducing the and PARAVAC projects continue to ensure
impact of disease or eliminating it alto- that expertise.
gether from a region or globally it is to
be noted that rinderpest was officially clas- Bluetongue virus (BTV) is spread by midges
sified as eradicated globally in 2011. It is (Culicoides species) and had been threat-
estimated that the cost of eradication/con- ening or present in southern Europe when
trol was USD 4 billion from 1945 to 2010. projects were commenced to investigate
Rinderpest previously caused devastation arboviruses. BTVAC was launched in 2007
to cattle farmers in Africa, the Middle East and Arbo-Zoonet followed on in 2009 as
and India and its eradication was greatly a means of spreading knowledge concern-
assisted by the development of a heat sta- ing arboviruses in the EU and outside. BTV
ble vaccine. With FMD, one of the goals of spread to northern Europe in 2006 and it
research is to develop a multivalent heat was a rush against time to produce inac-
stable vaccine that would allow us to fight tivate vaccines. However, vaccines were
this disease much more effectively as in the deployed to prevent a major outbreak of
case of rinderpest. the disease in 2009 that would have been
expected to lead to very considerable mor- EU has recognised the added benefit that
bidity and mortality in sheep and cattle can be achieved through coordination of
populations in Europe. The arbovirus work this activity using various instruments. The
has spread knowledge about these types European Technology Platform for Global
of diseases and has stimulated expertise in Animal Health (ETPGAH) was launched in
the veterinary entomology area some- 2004 as a stakeholder-driven initiative
thing that was greatly needed as very few with the objective of bringing a focus on
experts previously existed. the research effort across stakeholders.
A vision was developed and strategically
The VENoMYC project was launched in important issues identified. An action plan
2004 creating a large network of labora- then followed identifying 28 major actions.
tories working on mycobacterial diseases
(tuberculosis and paratuberculosis). The aim The action plan identified the need for a
was to lead to improved diagnostics, a bet- European research area network (ERA-NET)
ter understanding of the diseases, research in the animal health area to coordinate
on the use of vaccines, study of natural activity at the level of the research funders,
resistance, public health implications and and EMIDA was subsequently launched in
an identification of further research needs. 2009. This brought the funders of research
The TB-STEP project followed in 2008, from 19 countries together, leading to two
focusing only on tuberculosis to get a better calls for collaborative research across inter-
understanding of the epidemiology of the national borders, including some public-pri-
disease, the role of other species espe- vate partnerships. This represented a tre-
cially wildlife in the maintenance and mendous step forward in terms of research
transmission of the disease, utility of vac- synergy.
cines including in wildlife, better diagnos-
tics and to propose control strategies. It is The EMIDA ERA-NET is being followed by
important that this type of research contin- asecond ERA-NET with the short title of
ues as we struggle to control tuberculosis. ANIWHA, launched early in 2012.
The PCVD project in 2004 recognised the Another project that emerged from the
need to respond to the threat of porcine ETPGAH is DISCONTOOLS. This project
circovirus (PCV). The project achieved great focuses on disease prioritisation, with the
strides in terms of the use of vaccines to model being developed on the basis of the
control this very damaging disease. It was analysis of 51 diseases. Key information is
very interesting to see the synergistic role agreed leading to the identification of gaps
of nutrition as a factor in controlling the in knowledge or tools (diagnostics, vaccines,
disease. pharmaceuticals) and the diseases are then
prioritised by looking at our knowledge of
Later on, the NMSACC-PCVD project facili- the disease and its impact on animal health
tated the communication, networking and and welfare, public health, wider society,
training of all parties concerned in rela- trade and the availability of control tools.
tion to the control of PCV. There was also By having this prioritisation, we can focus
a particular focus on young scientists, new the research effort on the most critical gaps
Member States and accession countries. in the most important diseases.
Following the success of EMIDA, the STAR-

Synergy from EU-funded IDAZ project takes the concept of collabor
projects ation between funders on to a wider inter-
In many situations, there is considerable national level. STAR-IDAZ was launched in
research effort at the national level and the 2011, bringing Argentina, Australia, Brazil,
Canada, China, Europe, India, Mexico, New production due to endemic diseases,
Zealand, Russia and the United States to extremely high costs due to epidemic dis-
the table. The enthusiasm to collaborate eases and an ongoing threat to the human
exists as the big prize is the ability to control population due to zoonoses, investment in
disease, making everybody a winner in any research in this area makes good sense. We
advances that are achieved. This represents aim to reduce the impact of endemic dis-
a tremendous synergy in the research effort. eases this adds value not only in terms
of avoiding production losses but also con-
tributes to climate change mitigation by
Neglected zoonoses and using less inputs (feed, water, energy, etc.)
wildlife and producing less waste (manure, CO2 ,
The ICONZ project, launched in 2009, rec- methane, etc.). By preventing epidemic
ognises that a number of diseases cause a diseases via better control tools, we can
significant burden on poor and marginalised avoid significant costs to society as per
communities in Africa. The project focuses Table 1. Naturally, preventing zoonoses via
on endemic zoonotic diseases which have the development of better control tools
been identified by the World Health Organ has tremendous benefits to human health.
isation (WHO) as neglected. The diseases in The practical elimination of rabies across
scope include anthrax, brucellosis, bovine Europe via vaccines including vaccines
tuberculosis, cystic echinoccosis, leishman delivered to wildlife via baits is a very good
iasis, rabies, cysticercosis and trypanoso- example of a tremendous benefit to human
miasis. The project plans to add knowledge, health.
test culturally acceptable and cost-effec-
tive control interventions and engage By launching projects that ensure we have
with policymakers to seek better control the correct human capacity, by looking to
strategies. the future threats, by ensuring continu-
ity in the effort to develop solutions and
Research on animal diseases is expensive, by focusing on high-impact diseases, the
especially where high containment fac research programme over the past 10 years
ilities are required. The added advantage has been very well focused.
of linking existing research facilities and its
importance in underpinning the programme The initiation of the technology platform,
of research funded through the FAFB work the launch of networks and the develop-
programmes was recognised in the funding ment of ERA-NETS has ensured that the
of the European Network for Animal Dis- research effort becomes even more focused
eases and Infectiology Research Facilities and that it is approached in a collabora-
(NADIR) project by the Capacities, Research tive manner, building on previous work and
Infrastructures programme. This project avoiding overlap of effort.
unites the major experimental animal fac
ilities with capacity to undertake research The lateral thinking of also looking at wild-
on zoonoses, emerging diseases and other life reservoirs as a potential source of pre-
animal infectious diseases requiring bio- viously unknown diseases is an inspiring
containment at level 3 with the aim of shar- example of future planning. In addition, the
ing resources and a coherent development focus on neglected zoonoses ensures that
of high bio-containment facilities. the appropriate tools are provided to con-
trol these diseases of significant impact in
societies least able to respond.
Conclusions
Given the fact that animal diseases impose The research effort needs to continue. Dis-
a high impact on society in terms of lost eases continue to impact on animal health
and welfare and society at large. As we

move to a predicted global population of
9 billion by 2050, the time to develop new
and better tools to control disease is now.
We are making progress and continue to
do so. We must continue on the path that
we are on, moving towards an ever greater
focus on the priority research targets and
maximising synergy across all parties con-
cerned to make the research effort most
productive.
Declan O Brien
Managing Director IFAH-Europe.
Project Co-ordinator DISCONTOOLS
Alex Morrow
Coordination of research on animal health
Project Co-ordinator EMIDA ERA-NET
UK Department for the Environment Food
and Rural Affairs (DEFRA)
3.1. Mycobacterial diseases

[VENoMYC]
Veterinary network of
laboratories researching
into improved diagnosis
and epidemiology of
mycobacterial diseases
Summary network made up of 37 partners from Acronym:
Tuberculosis and paratuberculosis of live- 17countries. The partners was selected on VENoMYC
stock are mycobacterial diseases that rep- basis of active researching on mycobacte-
Project number:
resent a threat not only to public health rial infections. The work focused on seven 501903
but also have a high economic impact due tasks devoted to specific concerns, and
EC contribution:
to mortalities, condemnations, decreases in were divided into 11 work packages grouped
EUR 796 201
weight and fertility, and drop in productions. into three categories: (i) devoted to dis-
Countries of the European Union are under semination of knowledge to the Community; Duration:
60 months
eradication programmes for bovine tubercu- (ii)dedicated to specific harmonisations; and
losis and/or paratuberculosis. To date, eradi- (iii)focused on topics that will have a deep Start date:
cation has not been achieved in the EU due to impact on the understanding and control of 1 September 2004
several problems and, therefore, tuberculosis the diseases in the near future. Available Instrument:
and paratuberculosis remain major concerns tools for partners were general meetings, Coordination action
in livestock production. The most relevant workshops (lectures and hands-on training in
problems (lack of appropriate methods of the laboratory) and the exchange of person-
diagnosis, the role played in the epidemiology nel for training purposes. The main approach
of the diseases by other domestic and wild of this coordination action was to share tech-
animals, difficulties in the laboratory work nology and expertise in order to both avoid
with these pathogens, and lack of adequate research fragmentation and obtain a com-
vaccines that do not interfere with diagnosis) mon knowledge on mycobacterial diseases.
were addressed in this coordination action. The final target was to develop harmonised
In addition, the application of new systems recommendations and/or procedures for their
was discussed (i.e. use of functional genom- potential transposition into EU policies.
ics to detect new molecular markers and/or
to develop new vaccines).
Problem
The collaboration between the partners The main approach of this coordination
was structured through a multidisciplinary action is to share technology and expertise
in order to both avoid research fragmenta- WP8: Standardisation of molecular

tion and obtain a common knowledge on techniques for epidemiological
mycobacterial diseases. The final target studies;
was to develop harmonised recommenda- WP9: Wildlife reservoirs of mycobacterial
tions and/or procedures for their potential infections;
transposition into EU policies. WP10: A pplication of functional genomics
results on mycobacterial diagnosis;
The project brought together partners with WP11: U se of vaccines in the control of
expertise in a number of different technolo- tuberculosis and paratuberculosis.
gies for evaluation of the different diag-
nostic tests, control measures, molecular The strategic milestones of the VENoMYC
characterisation protocols, etc., for myco- coordination action were:
bacterial diseases. The project took advan-
tage of the partners experience in new accreditation of laboratories for the
technological developments to study the diagnosis of mycobacterial diseases and
application for tuberculosis and paratuber- approval of internal standards;
culosis diagnosis. description of a standard procedure to
implement the gamma-interferon test,
taking into account the current situation
Aim of European livestock (i.e. dual infection,
The key objective of this coordination action vaccination);
project was to develop a multidisciplinary application of typing methodology to
European network of laboratories research- disclose the relative importance of the
ing into mycobacterial diseases of veteri- sources of infection to understand the
nary interest. The strategic objective was epidemiology of the diseases;
the translation of the research results into recommended strategies to con-
EU policies. trol transmission of infections from
reservoirs to livestock;
The project was divided into seven possible uses of vaccines in the control
tasks and 11 work packages (WPs), of tuberculosis and paratuberculosis;
according to the different technologies understanding the advantages of appli-
employed by theparticipating labora- cation of functional genomics results on
tories. Someofthese tasks (WPs) were diagnosis;
an extension of tasks developed under evaluation of the public-health impli-
the project Concerted Action FAIR6- cations of mycobacterial infections in
CT98-4373. Theselected 11 work pack- animals.
ages were:
Expected results
WP1: Development and support of The expected outputs from this per-
VENoMYC website; manent network of researchers were:
WP2: Scientific audiovisual presentation; (i)improvement of the sensitivity of
WP3: General meetings; current tests and/or development of new
WP4: Laboratory diagnosis of diagnostic tests using new technologies
Mycobacterium spp.; which allow the early detection of
WP5: Direct detection of M. a. subclinical infections; (ii)molecular
paratuberculosis in dairy products; typing of mycobacteria; (iii)epidemi-
WP6: Immunology-based tests for ology studies; (iv)standardisation of
mycobacterial infections; techniques;(v)application of vaccines;
WP7: VNTR/MIRUs and DVR-spoligotyping (vi)study ofnatural resistance of cer-
for Mycobacterium bovis typing; tain species; (vii)potential public health
implications; and (viii)identification of obtained, reducing fragmentation between

research needs. research and diagnostic centres and refer-
ence laboratories.
Throughout the duration of the project,
two general meetings were organised: a Paratuberculosis and tuberculosis not only
first general meeting in 2004 and a final represent a significant disease of domes-
meeting in 2009. Moreover, a total of tic livestock but also of wildlife. The close
eight workshops were organised (Table1). communication between partners working
In the five years of the project, 31 short in different ecosystems enabled a greater
training mobilities were carried out. The level of understanding of the epidemiology
objectives of the short training mobility of the disease, and also represented a
funded within the VENoMYC project were diversity of countries with a wide variety of
to facilitate the inter-laboratory compari- wild animals.
son of techniques and to provide training
in specific methodologies. Attendance at Another advantage was that countries with
international colloquiums is an excellent a low prevalence in a mycobacterial infec-
opportunity to acquire updated knowl- tion, and therefore without active research
edge on all aspects related to mycobac- or specific facilities, received updated infor-
terial infections. For this reason, partners mation on the diagnosis and control of such
had also the opportunity to attend the infection that can be applied immediately in
international congresses on paratubercu- the event of an outbreak: this type of event
losis and M. bovis with sponsorship from is frequently associated with international
VENoMYC. trade of infected animals.
The outputs of WP7 (VNTR/MIRUs and DVR-

Potential applications spoligotyping for Mycobacterium bovis typ-
The project was best carried out at the ing) and WP8 (Standardisation of molecular
Community level because it brought techniques for epidemiological studies) con-
together expertise and exchange of tech- tributed to the definition of VNTR loci that
niques and methodologies that would not are recommended for typing of M.bovis,
be available within any one single Member spacers for an enhanced discriminative
State. For example, a European dimen- typing for M. bovis and enhanced use of
sion is needed to standardise procedures databases for mycobacterial typing.
for molecular characterisation techniques,
and to obtain the appropriate variety of The results obtained in WP6 (Immunology-
strains required to carry out epidemio based tests for mycobacterial infections)
logical studies at a European level. This contributed directly in the field implemen-
allowed the direct comparison of results tation of the IFN- assay by the European
Table 1: List of workshops organised within the VENoMYC project
DATE PLACE WORKSHOP WORKSHOP

2629 Jena, WP4 Laboratory diagnosis of Mycobacterium
April 2006 Germany spp.
1316 Maynooth, WP9 Wildlife reservoirs of mycobacterial
September 2006 Ireland infections
1921 Toledo, WP7 VNTR/MIRUs and DVR-Spoligotyping for
October 2006 Spain Mycobacterium bovis typing
DATE PLACE WORKSHOP WORKSHOP

912 Laguardia, WP11 Use of vaccines in the control of
November 2006 Spain paratuberculosis and tuberculosis
1013 May 2007 Athens, WP5 and WP8 Detection of M. a. paratuberculosis in dairy
Greece products and Standardisation of molecular
techniques for epidemiological studies
1619 Cambridge, WP10 Application of functional genomics results
September 2007 United on mycobacterial diagnosis
Kingdom
30 and 31 Belfast, WP6 Immunology-based tests for mycobacterial
May 2008 United diseases
Kingdom
2325 Madrid, WP7 VNTR/MIRUs and DVR-Spoligotyping for
March 2009 Spain Mycobacterium bovis typing Results of
the VNTR-DVR Spoligotyping Ring Trial
Commission. Moreover, the description of news; and

transmission of tuberculosis between wild- links to international organisms on
life and livestock derived from WP9 (Wildlife animal health, databases, etc.
reservoirs of mycobacterial infections) has
an impact on the control of the disease if Keywords
staff of Ministries of Agriculture and Fisher- tuberculosis, paratuberculosis, M. avium
ies and Environment take them into account complex, Mycobacterium bovis, Mycobac
for the design of eradication campaigns. terium caprae, Mycobacterium avium
subsp. paratuberculosis, standardisation,
epidemiology, wildlife, vaccines, diagnosis,
References/publications epidemiology, genomics
The VENoMYC coordination action did not
fund research activities but at each general
meeting and workshop organised within the
VENoMYC project, a book of proceedings
was handed to all participants (downloaded
from the project website). Moreover, stand- VENoMYC logo
ardised protocols for typing mycobacteria
were also available for all partners. Partnership
The consortium was designed to include
Project website at least one Institution from each Euro-
The website (http:///www.venomyc.com) pean country to ensure a representative
was named after the acronym of the pro- image of problems caused by mycobacte-
ject: VENoMYC (Veterinary European Net- rial infections. The fact that some countries
work of Mycobacteria). Spain, Italy and the United Kingdom) were
over-represented reflects a higher impact
Information on the website includes: of mycobacterial diseases and active
research. The needed link with human
objectives of the coordination action; impact was obtained with integration of
work packages; partners P19 (RIVM, Netherlands) and P24
a list of partner institutions; (AGU, Greece).
contact details;
meeting plans (workshops information The scientific component of the Consortium
and reports); (37 partners) created a multidisciplinary
team integrating disciplines such as disease Partner 1, UCM (Coordinator)

diagnosis, epidemiology, typing and control Dr Lucas Domnguez
over 17 European countries. The partner- Departamento de Sanidad Animal
ship included a combination of universities, Facultad de Veterinaria
research institutes and reference labora- Avda. Puerta de Hierro, s/n
tories. The partners complemented one 28040 Madrid
another to ensure collaboration with groups SPAIN
in Europe working with mycobacterial Tel. +34 913943721
diseases. Fax +34 913943795
E-mail: lucasdo@visavet.ucm.es
Interaction between 60 % of the partici-
pant laboratories began with the previous
EU programme FAIR6-CT98-4373, Con-
certed action for the setting-up of a Euro-
pean veterinary network on diagnosis, epi-
demiology and research of mycobacterial
diseases.
Partners
ACRONYM PARTNER LEADER ORGANISATION
UCD Dr Eamonn Gormley University College of Dublin, Ireland
CVRL Dr Eamonn Costello Department of Agriculture and Food,
Ireland
QUB Dr Sydney Neill Queens University Belfast, United
Kingdom
MRI Dr Karen Stevenson Moredun Research Institute, United
Kingdom
AFSSA Dr Maria Laura Boschiroli Agence franaise de securit sanitaire
des aliments, France
SSI Dr Peter Andersen Statens Serum Institut, Denmark
DVI Dr Peter Ahrens Danish Institute for Food and Veteri-
nary Research, Denmark
CIDC-Lelystad Dr Douwe Bakker Central Institute for Animal
Disease Control, Netherlands
SVA Dr Gran Blske Statens Veterinrmedicinska
Anstalt,Sweden
NVI Dr Berit Djnne National Veterinary Institute, Norway
VRI Dr Ivo Pavlik Veterinary Research Institute, Czech
Republic
IZSPLV Dr Maria Goria Istituto Zooprofilattico Sperimentale
del Piemonte, Liguria e Valle dAosta,
Italy
UEx Dr Javier Hermoso-de Mendoza Universidad de Extremadura, Spain
SGHMS Dr Tim Bull St Georges Hospital Medical School,
United Kingdom
NAGREF Dr Zoi Dimarelli National Agricultural Research Founda-
tion, Greece
ACRONYM PARTNER LEADER ORGANISATION

EELA Dr Jaana Seppnen National Veterinary and Food Research
Institute, Finland
VETHS Dr Anne Storset The Norwegian School
of Veterinary Science, Norway
RIVM Dr Dick Van Soolingen National Institute for Public Health and
the Environment, Netherlands
BFAV Dr Heike Khler Federal Research Centre for Viruses
Disease of Animals, Germany
IZSLT Dr Eugenio Lillini Istituto Zooprofilattico Sperimentale
Lazio e Toscaza, Italy
LNIV Dr Alice Amado Laboratrio Nacional de Investigao
Veterinria, Portugal
VAR Dr Karl Walravens Veterinary and Agrochemical Research
Centre, Belgium
AGU Dr Ioannis Ikonomopoulos Agricultural University of Athens,
Greece
UVM Dr Milan Travnicek University of Veterinary Medicine,
Slovakia
SAC Dr Mike Hutchings Scottish Agricultural College, United
Kingdom
NEIKER Dr Ramn Juste Instituto Vasco de Investigacin
y Desarrollo Agrario, Spain
IZSLER Dr Maria Pacciarini Istituto Zooprofilattico Sperimen-
tale della Lombardia e dell Emilia
Romagna, Italy
LCSA-SANTA FE Dr Fulgencio Garrido Laboratorio Central de Sanidad Animal
del Ministerio de Agricultura, Pesca y
Alimentacin en Santa Fe, Spain
VLA Dr Glyn Hewinson Veterinary Laboratories Agency, United
Kingdom
IPL Dr Philip Supply Institut Pasteur de Lille, France
VF Ljubljana Dr Milan Pogacnik Veterinary Faculty University of Lju-
bljana, Slovenia
UNILEON Dr Juan F. Garca-Marn Universidad de Len, Spain
TiHo Dr Gerald Gerlach School of Veterinary Medicine Han-
nover, Germany
UNISS Dr Leonardo A. Sechi Universita degli studi di Sassari, Italy
UM Dr Luis Len Vizcano Universidad de Murcia, Spain
INRA-918 Dr Laurence Guilloteau Institut national de la recherche
agronomique, France
[TB-STEP]
Strategies for the eradication

of bovine tuberculosis
Summary made up of 12 partners from eight coun-
Acronym:
Tuberculosis is an infectious disease caused tries that will carry out research on seven TB-STEP
by microorganisms of the Mycobacterium work packages devoted to improved tools
Project number:
tuberculosis complex. This infection affects and to developing strategies for the eradi-
212414
domestic and wild animals and represents cation of bovine tuberculosis in areas where
a major concern worldwide because of its the disease is present in both domestic and EC contribution:
EUR 2 894 759.00
high economic impact due to mortalities, wildlife populations. It will include: (i)vac-
condemnations, decreases in productions, cination of bovine animals and wildlife, Duration:
and its zoonotic potential. (ii)control of populations, to reach numbers 39 months
compatible with animal welfare, and strat- Start date:
Eradication programmes based on a test- egies to limit contact between domestic 1 October 2008
and-slaughter policy have been imple- and wild species; and (iii)development of Instrument:
mented in the European Union, and have improved diagnostic tools for detection of Collaborative project
proved successful in some countries. How- infected animals.
ever, they have been unable to eradicate
the infection in others despite the use
of vast economical resources. A relevant Problem
problem is the existence of infected wild- The Mycobacterium tuberculosis com-
life: the best known examples are the plex causes pulmonary, gastrointestinal
European badger (Meles meles) in United and disseminated disease in mammals.
Kingdom and Ireland, and the wild boar M. bovis and M. caprae are the most rel-
(Sus scrofa) in Spain. Besides this fact, evant in animal health. M. bovis has an
there is only a limited knowledge about exceptionally large host range, including
other potential underlying causes, such as: domestic animals and wildlife. Tuberculo-
(i)the real contribution of cattle-to-cattle sis transmitted between wildlife, livestock
transmission at the same area (neighbour- and humans presents major challenges for
ing farms and communal pastures) or after the protection of human and animal health,
movement of animals; (ii)the role played the economic sustainability of agriculture
in the epidemiology by other domestic ani- and the conservation of wildlife. The poten-
mals; or (iii)the effect of interferences in tial for non-ruminants and wildlife to act
the diagnosis tests. The weight of these as reservoirs has opened up new lines of
causes may also differ depending on the research in the epidemiology of mycobac-
farming system and ecological factors. It terial infections. Therefore, it is important
is unlikely that there can be a single solu- to determine the role of wildlife in the epi-
tion as it is unlikely that there is a single demiology of tuberculosis in domestic live-
cause. stock to establish control strategies that
are pertinent to the range of agricultural
Aiming at the eradication of this infection, systems employed in the EU. The Euro-
this TB-STEP project has planned a mul- pean badger (Meles meles) is implicated in
tifaceted battlefront. The consortium is the transmission of M. bovis to cattle and
constitutes the most important reservoir the project derives from the two keywords
of infection in the United Kingdom and in the idea: the design of strategies based
Ireland. The wild boar (Sus scrofa) seems on a sound understanding of the epidemi
to be the main reservoir for tuberculosis in ology (TB-STEP). To approach the eradi-
Spanish Mediterranean habitats. M. bovis cation of this infection, this project plans
is also a threat to valuable wildlife species a multifaceted battlefront because the
and exotic animals. hypothesis is that a combination of meth-
odologies would be needed to achieve the
task. Thus, research is focused on topics
that will have a deep impact on the under-
standing and control of the diseases in
the near future. The work packages have
been designed to share technology and
expertise in order to both avoid research
fragmentation.
More specifically, the project will:
identify the relative impact of the

different potential causes in the
epidemiology;
understand the role of other species
in the epidemiology (maintenance and
transmission);
study the effect of vaccination in cattle
Thus, the eradication of tuberculosis would and goats;
require eradication from domestic animals assess the safety and immunogenicity
and control (or eradication) of the infec- of vaccine in wildlife;
tion in wildlife. However, to date this has study the suitability of vaccines in
been hampered by a number of reasons: wildlife;
(i)the limited sensitivity and specificity of develop affordable serological tests to
the in vivo diagnostic test in cattle under use in wildlife to estimate prevalence;
some circumstances; (ii)the lack of diag- improve cattle diagnosis by detecting
nostic tests in wildlife, making difficult it to interferences;
estimate the prevalence and the high-risk develop a model system to assess the
areas; (iii)the difficulties in using vaccines risk, taking into account the ecological
regarding the cross-reactions (cattle) and and farm management factors;
delivery (wildlife), as well as the limited design the optimal strategy for control
protection; and (iv)the insufficient under- of infection in wildlife.
standing about the relative contributions
of each (domestic and wild) species and Expected results
the impact of the management practices. The final target is to develop rational strat-
The four major points presented above are egy/ies and/or procedures for their poten-
addressed by this TB-STEP project. tial transposition into EU policies. For that
reason, the project develops a multidis
ciplinary approach ranging from vaccina-
Aim tion and immunology, molecular epide-
The project aims at the design of strategies miology, diagnosis to ecology. TB-STEP
to achieve the eradication of tuberculosis contributes to the scientific, technical,
from livestock and wildlife. The acronym of social and policy objectives (Cooperation
Theme 2 Food, Agriculture and Fisheries, Thacker, T.C., MeIjlis, J., Carter, C., Gordon,
and Biotechnology (European Commission S., Egnuni, T., Hardegger, R., Marg-Haufe,
C(2006) 6839)). The results from the TB- B., Raeber, A., Oesch, B., Comparison
STEP project will lead to greater efficiency of tuberculin activity in the interferon-
and cost savings which will improve the gamma assay for the diagnosis of bovine
competitiveness of the European livestock tuberculosis, Veterinary Record, 167(9):
and dairy industries in internal and global 322326, 2010.
markers. Furthermore, it would contribute
to the demand for safer food, the sustaina- Acevedo, P., Ferreres, J., Jaroso, R., Durn,
ble use of and production of bio-resources, M., Escudero, M.A., Marco, J., Gortzar, C.,
the risk of zoonoses diseases and concern Estimating roe deer abundance from pel-
on animal welfare. let group counts in Spain: An assessment
of methods suitable for Mediterranean
The project is also contributing to building woodlands, Ecological Indicators, 10(6):
a European knowledge-based bio-economy 12261230, 2010.
as it brings together science (the research
organisations), industry (the SMEs) and Rodrguez, S., Bezos, J., Romero, B., de
stakeholders (policymakers). Juan, L., lvarez, J., Castellanos, E., Moya,
N., Lozano, F., Javed, M.T., Sez-Llorente,
J.L., Libana, E., Mateos, A., Domnguez,
Potential applications L., Aranaz, A., The Spanish Network on
The project will deliver strategies to be Surveillance and Monitoring of Animal
applied in the tuberculosis eradication Tuberculosis, Mycobacterium caprae
campaigns in the affected EU countries infection in livestock and wildlife, Spain,
where the disease is present in both Emerging Infectious Diseases, 17(3):
domestic and wildlife populations. Poten- 532535,2011.
tial applications include: (i)vaccination of
bovine animals, wildlife and feral reser- Bhm, M., Hutchings, M.R., White, P.C.L.,
voirs; (ii)control of populations to reach Contact networks in a wildlife-livestock
numbers compatible with animal welfare; host community: identifying high-risk indi-
(iii)improved diagnostic tools for detection viduals in the transmission of bovine TB
of infected animals; (iv)strategies to limit among badgers and cattle, PLoS One, 4,
the contact between domestic and wild 112, 2009.
species. The diversity of wild species (some
legally protected) and farming systems are Project website
also taken into account. http://www.vigilanciasanitaria.es/tb-step/
Keywords
References/publications tuberculosis, Mycobacterium bovis, Myco-
Ballesteros, C., Garrido, J. M., Vicente, J., bacterium caprae, strategies, wildlife,
Romero, B., Galindo, R.C., Minguijn, E., Vil- vaccines, diagnosis, epidemiology
lar, M., Martn-Hernando, M.P., Sevilla, I.,
Juste, R., Aranaz, A., de la Fuente, J., Gor-
tzar, C., First data on Eurasian wild boar
response to oral immunisation with BCG
and challenge with a Mycobacterium bovis
field strain, Vaccine, 27: 66626668, 2009.
Schiller, I., Vordermeier, H.M., Waters,

W.R., Cagiola, M., Whelan, A., Palmer, M.V., TB-Step Logo
Partnership The partnership will establish collaboration

The collaboration between the partners with researchers from developing countries
will be structured through a multidiscip (from the ICPC). Potential candidates have
linary network made up of 12 partners been identified from countries located in
from seven European countries (Hungary, Africa and South America, based on known
Ireland, Italy, the Netherlands, Spain, Swit- interest in these areas of animal health.
zerland, and United Kingdom), and a third
country (South Africa). The fact that some
countries (ES, UK) are over-represented Coordinators
reflects a higher impact of mycobacte- Dr Lucas Domnguez (Project Coordinator)
rial diseases and also the reported role Dr Luca de Juan (Deputy Coordinator)
of wildlife as reseroirs of the infection. In VISAVET Animal Health Surveillance Centre
total, 10research organisations and two Faculty of Veterinary Sciences
SME (Ingenasa and Prionics) are involved in Universidad Complutense de Madrid (UCM)
the Consortium. The partnership includes a SPAIN
combination of universities, research insti- lucasdo@visavet.ucm.es
tutes and Reference Laboratories. dejuan@visavet.ucm.es
Partners
Instituto de Investigacin en Dr Christian Gortazar christian.gortazar@uclm.es
Recursos Cinegticos, Wildlife
Disease Department (IREC),
Universidad de Castilla-La
Mancha (UCLM),
SPAIN
Badger Vaccine Research Dr Eamonn Gormley egormely@ucd.ie
Team, School of Agriculture,
Food Science and Veterinary
Medicine (UCD) Dublin
IRELAND
Veterinary Laboratories Dr Glyn Hewinson Glyn.Hewinson@ahvla.gsi.gov.uk
Agency, TB Research Group
(VLA)
Department for Environ-
ment, Food and Rural Affairs
(Defra)
UNITED KINGDOM
Scottish Agricultural College Dr Michael Hutchings mike.hutchings@sac.ac.uk
(SAC)
West Mains Road
Edinburgh
UNITED KINGDOM

Veterinary Sciences Division, Dr James McNair jim.mcnair@afbini.gov.uk
Agri-Food and Biosciences
Institute (AFBINI)
Queens University of Belfast
(QUB)
Belfast
UNITED KINGDOM
Laboratory for Mycobacterial Dr Douwe Bakker douwe.bakker@wur.nl
Infections
Department of Bacteriol-
ogy and TSEs of the Central
Veterinary Institute (CVI)
Lelystad
NETHERLANDS
Istituto Superiore di Sanit Dr Paolo Pasqualy paolo.pasquali@iss.it
(ISS)
Rome
ITALY
Laboratory of TB and Clinical Dr Szilrd Jnosi janosis@oai.hu
Bacteriology
Veterinary Diagnostic Direc-
torate (VDD)
Mezogazdasagi Szakigaz-
gatasi Hivatal Kozpont or
Central Agricultural Office
(CAO)
HUNGARY
Ingenasa Dr Carmen Vela cvela@ingenasa.es
SPAIN
Prionics AG Dr Alex Raeber Alex.Raeber@prionics.com
SWITZERLAND
Faculty of Veterinary Science Dr Anita Michel anita.michel@up.ac.za
of the University of Pretoria
(UP)
Onderstepoort
SOUTH AFRICA
3.2. Parasites
[DELIVER]
Design of effective and

sustainable control strategies
for liver fluke in Europe
Summary EU Member States. In the United Kingdom,
Acronym:
DELIVER
The DELIVER project aimed to improve the prevalence of infection in cattle ranges
methods for controlling liver fluke disease. from 45 % to 84% and in Ireland alone,
Project number:
The project started in 2006 in response annual losses have been estimated at over
023025
to growing concerns in Europes agricul- EUR 60 million. Control of the disease in
EC contribution: tural industry about the impact of the liver livestock is based on the use of anthel-
EUR 3 540 000
fluke parasite on productivity. This parasite, mintic drugs, with accompanying risks of
Duration: Fasciola hepatica, infects cattle, sheep, chemical residues in foodstuffs. Moreover,
40 months goats and other exotic farmed species anthelmintic resistance is emerging as
Start date: that eat grass. It has a complex life cycle aproblem in many areas of Europe and glo
1 February 2006 that is dependent on suitable temperature bally. Fasciolosis is also an emerging human
Instrument: and rainfall and involves an intermediate disease in many INCO countries, with an
Specific targeted snail host. The main areas of concern are: estimated 17million people infected.
research or innovation (i)changing climate that better suits the
project (STREP) parasites life cycle and allows it to spread Current control of fasciolosis in ruminants
to new areas; (ii)resistance to treatment is based on prophylactic/therapeutic use
with the widely used flukicide drug tricla- of flukicidal (anthelmintic) drugs, with
bendazole; (iii)the parasites ability to make the risk that residues of these drugs may
animals more susceptible to other bacterial contaminate food and the environment.
infections such as salmonellosis and bovine The DELIVER project proposes to enhance
tuberculosis. The DELIVER project has been the safety and quality of the food supply
successful in bringing together 15 research available to European consumers by devel-
groups from Europe and Latin America oping improved, environmentally-friendly
to tackle these problems and provide methods for the control of this infection in
solutions. domestic livestock. Improved control will
also decrease the potential for transmis-
sion to humans of this infection via plant
Problem products.
Liver fluke disease causes annual losses of
over EUR 2.5 billion to livestock production
and the food industry worldwide. The preva- Aim
lence of fasciolosis is dramatically increas- The main objective of the project was
ing; in recent years, for example, increases to develop novel control methods for
of up to twelvefold have been recorded in fasciolosis in livestock, thus enhancing
food safety and allaying consumer con- Results

cerns. This objective included three areas Epidemiology: Over 4 000 samples of milk
of work. and serum were collected from England,
Greece, Spain and Wales to determine the
E pidemiology: The aim of the work
1. prevalence of infection in livestock in these
in this package was to improve our countries. The large-scale survey in England
understanding of the epidemiology of and Wales has provided a complete cover-
fasciolosis by determining farm-spe- age of these countries and enabled us to
cific risk factors for infection, the role construct a GIS showing the spatial distri-
of wildlife in transmission, improving bution of F. hepatica. A risk analysis showed
the sensitivity of diagnosis of patent that climatic variables, particularly rain-
infection and improving our under- fall, temperature, soil type and presence
standing of how genetic variation in of sheep are significantly correlated with a
populations of fluke affect their inter- high prevalence of exposure to F. hepatica
action with the intermediate snail host at a postcode area level. Interestingly, the
and their infectivity and pathogenic- data suggests that climatic variables occur-
ity for farmed livestock. Ultimately, ring in the previous year or over a five-year
these results will lead to web-based period can be used to determine the risk of
predictive models which farmers can F. hepatica infection. However, farm-specific
use to assess the seasonal and year- factors have yet to be identified to account
on-year risk of infection and disease for variation between individual farms in
in their herds and flocks and assist in close proximity.
decision-making to assess if treatment
is necessary, when to treat and which Characterisation of the F. hepatica soluble,
drug to use. This work package also juvenile and egg proteomes has been com-
determined the significance of fluke pleted. A novel GST has been identified and
infection in humans within the EU. characterised and, in addition, an antigenic
A nthelmintic resistance: The main
2. high molecular weight HSP (FhHEA1) iden-
aims were: standardisation of field and tified in eggs. A host trypsin inhibitor com-
in vitro assays for detecting anthel- plex has been identified as a biomarker of
mintic resistance and determining the infected animals. A diagnostic PCR has been
extent of triclabendazole resistance in developed that can detect a single egg per
Europe and in INCO partner countries; gram of faeces in naturally infected animals.
and determination of mechanisms of An analysis of a panel of sera from a group
drug action and resistance in order to of farmworkers considered to be a high risk
formulate new strategies for conser- population, showed no evidence that they
vation of drug efficacies. This work had been exposed to F. hepatica infection.
package also aims to develop protoc
ols for the measurement of drug RFLP analysis of mitochondrial DNA from
residues in food products and in the flukes has provided markers that have been
environment. used to assess the genetic diversity of
I mmunoprophylaxis: This work aims
3. flukes of known provenance, isolated from
to provide alternative, immunoprophy- different regions of the EU. Initial results
lactic means of controlling fasciolosis. suggest that there is extensive diversity
Work towards this aim encompasses between isolates that is not geographically
studies of the basic immunology of restricted. This raises questions about
infection as well as vaccine trials with potential differences in the virulence and
new and existing recombinant anti- infectivity between different isolates and
gens, and with DNA, carried out under also the potential for the emergence and
controlled conditions. spread of drug resistance through Europe.
Drug Action: Anthelmintic drugs are of this effect was obtained using a model
still the commonest means of controlling system of co-infection with F. hepatica and
fluke infection. Over the last 20 years, tri- Mycobacterium bovis in cattle.
clabendazole has been the drug of choice
for many livestock farmers, but cases of The difficulties inherent in carrying out vac-
resistance have been reported in veterinary cine trials, and the compounding factors
journals in the United Kingdom and other that can affect the interpretation of results,
European countries. Research groups from became clear during this period. Preliminary
Europe and Argentina have been collaborat- evidence of differences in innate suscep-
ing through the DELIVER project to try and tibility to infection in sheep was obtained,
understand how triclabendazole kills fluke and an adjuvant effect using Quil A, again
and how resistance develops. in sheep, was demonstrated. Several vac-
cine trials, using various combinations of
One success has been the finding that the antigens, did not demonstrate any consist-
effectiveness of drug can be increased ent protective effect. A field trial in cat-
by co-treatment with appropriate meta- tle indicated that direct neutralisation of
bolic inhibitors. We now have a much bet- CL1activity by antibodies from immunised
ter understanding of how the parasite animals is not likely to be a major protec-
resists drug action and this may lead to new tive mechanism. Overall, the results of this
therapies to improve the efficacy of the drug. period emphasise the difficulties of vaccine
trials in target species, while highlighting
Measuring drug resistance: Surveys in some aspects of the nature of the host-
Ireland, the Netherlands and the United parasite relationship. These findings ill be
Kingdom have provided data on how important in the path to an effective vaccine.
widespread resistance to triclabendazole
is. In Ireland, resistance was present on
three out of six farms investigated. A test Potential applications
to measure reduction in fluke egg count fol- The DELIVER project has been very success-
lowing treatment is being developed and ful in meeting its objectives by improving
standardised by the Veterinary Laboratories our understanding of the parasite in these
Agency in England and Wales, and in Ireland key areas. This is important because once
at the Teagasc Sheep Research Farm. a parasite population has acquired drug
resistance, there is no reversion back to
Immunoprophylaxis: During the pro- susceptibility even if the drug is withdrawn.
ject, significant advances were made in
understanding the immune response, the If the liver fluke affects the susceptibility of
mechanisms by which the parasite exerts cattle and sheep to other infections such as
a regulatory effect on the immune sys- bovine TB, control of the parasite will have
tem, and the relevance of this effect for benefits for the control of other infections
livestock health and infection with other as well. Progress has been for developing a
pathogens. Signifcant advances were also protective vaccine in cattle.
made; particularly through the use of prote
omics and using mouse models where it Finally, we have identified key risk factors
was demonstrated that tegumental anti- both at the individual farm level and at an
gen of F. hepatica actively suppressed Th1 area level that can be used to measure the
responses, through interaction with den- likely risk and severity of liver fluke disease
dritic cells. These findings are relevant to occurring each year. These will form the basis
the compromised ability to control bacte- of an online forecasting system that will be
rial infections seen in fluke-infected ani- available to farmers within the course of the
mals. Evidence of the real-life importance next few years. The effect of climate change,
the spread of drug resistance and changing Project website

patterns in EU agriculture are likely to make http://www.deliver-project.eu/
the economic impact of liver disease even
more significant in the next decade. Keywords
liver fluke, Fasciola hepatica, epidemiology,
The research results have been used to anthelmintic resistance, vaccine, immune
produce guidelines for liver fluke control response
that has been distributed to farmers and
veterinarians organisations to help curb the Coordinator
spread and severity of the disease leading Prof. Jos Prez
to healthier animals and a better return for Universidad de Crdoba
farmers. Facultad de Veterinaria
Edificio Sanidad Animal
Campus de Rabanales
References/publications Ctra. Madrid-Cdiz km 396
Publications supported by the project 14014 Crdoba
can be obtained from the project web- SPAIN
site (http://www.deliver-project.eu/index. an1pearj@uco.es
php?page=information-for-researchers).
Partners
Dublin City University Dr Sandra ONeill sandra.oneill@dcu.ie
School of Nursing
Dublin
IRELAND
Universitary College Dublin Prof. Grace Mulcahy grace.mulcahy@ucd.ie
School of Agriculture
Food Science and Veterinary Medicine
Dublin
IRELAND
University of Wales Aberystwyth Prof. Peter Brophy pmb@aber.ac.uk
Biological Sciences
Faculty of Science
Aberystwyth
UNITED KINGDOM
University of Exeter Prof. Jennifer A. J.A.Littlechild@ex.ac.uk
Henry Wellcome Centre for Biocatalysis Littlechild
Devon
UNITED KINGDOM
Liverpool School of Tropical Medicine Prof. Diana Williams williadj@liv.ac.uk
Liverpool
UNITED KINGDOM
Witold Stefanski Institute of Parasit Prof. Halina halinwe@twarda.pan.pl
ology Polish Academy of Sciences Wedrychowicz
Warsaw
POLAND

Queens University Belfast Prof. Ian Fairweather i.fairweather@qub.ac.uk
School of Biological and Food Sciences
Belfast
UNITED KINGDOM
Instituut voor Dierhouderij en Dr Fred H.M. fred.borgsteede@wur.nl
Diergezondheid BV Borgsteede
Lelystad
NETHERLANDS
Agricultural University of Athens Dr Georgios gtheo@aua.gr
Faculty of Animal Science Theodoropoulos
Athens
GREECE
Institute of Experimental Pathology and Dr Ilia Bankov i_bankov@hotmail.com
Parasitology BASc
Sofia
BULGARIA
Universidad Nacional del Centro de la Dr Carlos E. Lanusse clanusse@vet.unicen.edu.ar
Provincia de Buenos Aires
Facultad de Ciencias Veterinarias
Laboratorio de Farmacologa Veterinaria
Tandil
ARGENTINA
Oswaldo Cruz Foundation Dr Miriam Tendler mtendler@ioc.fiocruz.br
Department of Helminthology
Fiocruz
BRAZIL
Universidad Nacional de Cajamarca Dr Pedro Ortiz ortizpedroluis@yahoo.es
Facultad de Veterinaria
Cajamarca
PERU
Lara Media Ltd Gerald Cannon ger@laramedia.net
Dublin
IRELAND
University of Liverpool Prof. Peter Brophy pmbrophy@liv.ac.uk
School of Biological Sciences
Liverpool
UNITED KINGDOM
Instituut voor Dierhouderij en Dr Fred H.M. fred.borgsteede@wur.nl
Diergezondheid BV Borgsteede
Animal Sciences Group of Wageningen
UR
Lelystad
NETHERLANDS
[PARASOL]
Novel solutions for the

sustainable control of
nematodes in ruminants
Summary conditions, for identifying animals that Acronym:
The overall objective of PARASOL was to require treatment, and produced and stand- PARASOL
create lowinput and sustainable strategies ardised tests for anthelmintic resistance to
Project number:
for the control of gastrointestinal nema- ensure that the drugs remain effective. 022851
tode infections of ruminants. These para-
EC contribution:
sites pose the greatest current threat to We produced clear guidance and proto-
EUR 2 940 000
agricultural productivity and animal welfare. cols for sustainable, low-input, user- and
consumer-friendly nematode control. To do Duration:
40 months
this, we:
Problem Start date:
Gastrointestinal nematodes pose the great- (i) assessed the effect of TSTs on product 1 February 2006
est current threat to agricultural productiv- ivity, animal welfare and the spread of Instrument:
ity and animal welfare. Current conventional anthelminthic resistance (AR) genes Specific Targeted
methods for worm control involve repeated under a range of farming conditions; Research or
Innovation Project
dosing of whole herds with synthetic anthel- (ii) determined the best methods of iden- (STREP)
mintics. This approach is not sustainable tifying animals and herds requiring
since it promotes the spread of anthelmintic anthelmintic intervention;
resistance by failing to leave untreated para (iii) standardised existing in vivo and in vitro
sites in refugia and there are concerns with tests for detecting AR and developed
regard to food residues and environmental new tests where previous ones were
impact. Nevertheless, effective chemical inadequate;
anthelmintics remain irreplaceable for worm (iv) tested the potential for optimising the
control, and their elimination is not practical efficacy and bioavailability of anthel-
on animal welfare and economic grounds. mintics by modulating parasite P-glyco-
protein detoxification systems;
(v) communicated with farmers, veterinar
Aim ians, advisors and trained animal health
The lack of a sound scientific basis for tar- technicians throughout the participat-
geting anthelmintic treatments has meant ing countries, to produce and dissemin
that producers have become over-reliant ate guidance to ensure good uptake
on intensive chemoprophylaxis. With the and implementation of the protocols
PARASOL project, we hope to replace cur- produced.
rent practice with targeted selective treat-
ments (TSTs), where only animals showing
clinical symptoms or reduced productiv- Expected results
ity are given drugs. We assessed several The overall objective of PARASOL work
innovative methods, under various farming packages 1 and 2 was to provide a scientific
rationale for deciding which animals to treat. that the egg hatch test can be used for
PARASOL studies have confirmed the value surveillance for benzimidazole resistance
of the TST approach in maintaining accept- in bovine nematodes but further research
able levels of productivity and conserving is required before the larval development
anthelmintic efficacy. The ability to target test is reliable for use with cattle worms.
treatments developed in the PARASOL pro-
ject has greatly facilitated the development Protocols for the running of FECRTs in
of decision support systems and enabled sheep have been drawn up and combined
researchers to make regionally appropriate with advice on the conducting field trials for
recommendations and guidelines. resistance surveys. Work package 6 exam-
ined the possibility of enhancing drug bio-
These are invaluable for farmers, advisors availability by targeting the mechanisms of
and policymakers who are concerned with drug transport and metabolism, in an effort
improving sustainability, minimising tissue to conserve the efficacy of our current
residues and the environmental impact anthelmintic families.
of anthelmintic control strategies. As
expected, the research conducted within The research conducted within PARASOL
PARASOL has confirmed the need for rec- has highlighted the problem of anthelmin-
ommendations that are tailored to fit the tic resistance in ruminants and has lead
local prevalence of the different economi- to the production of regionally specific
cally important parasites, the situation recommendations, based on the following
with regard to anthelmintic resistance, and recommendations.
regional production systems. The findings
from work packages 1 and 2 have been Targeted selective treatment (TST)
disseminated locally to farmers and more and targeted treatment (TT) strategies
widely to advisors, the major research should be promoted to enable effect
providers and policy-makers. Work pack- ive and sustainable worm control in
age 3 was specifically designed to provide ruminants.
standard tests for the detection of anthel- Anthelmintic efficacy should be routinely
mintic resistance as an invaluable adjunct monitored.
to work packages 1 and 2. Field surveys
using the Faecal Egg Count Reduction Test Evidence from the PARASOL research find-
(FECRT) in cattle have shown macrocyclic ings highlighted the need for an ongoing
lactone-resistant Cooperia are present in research effort to:
Belgium, Germany, Sweden and the United
Kingdom and Ostertagia in the United develop new in vitro and molecular tests
Kingdom. Because of the importance of for monitoring efficacy and potential
the cattle industry to the EU, continued resistance;
surveillance for anthelmintic resistance is provide a better understanding of the
recommended and strategies to slow the mechanisms of drug resistance;
development and spread of resistance are develop solutions for reversal of
required. The research in work packages resistance and improved drug efficacy.
4 and 5 was ultimately aimed at provid-
ing additional standard methodologies for Potential applications
the detection of anthelmintic resistance, The findings and outputs from all of these
particularly to anthelmintics in the macro work packages will have a considerable
cyclic lactone family. These tests will be strategic impact on livestock management
used as the basis of standardised proto- in Europe and selected INCO countries. The
cols for conducting surveys into the extent dissemination of these results to farmers
of anthelmintic resistance. It was shown and their advisors will allow them to make
rational decisions regarding the optimal Project website

use of anthelmintics within their region and http://www.parasol-project.com
production systems. Overall, the PARASOL
project has provided an improved under- Keywords
standing of the development and detection gastrointestinal nematodes, ruminants,
of anthelmintic resistance and the mech anthelmintic resistance, target selective
anisms that underpin it. This knowledge will treatment, nematode control, anthelmintic
contribute to other investigations into AR, resistance mechanisms
particularly human parasites.
Coordinator
Jozef Vercruysse
References/publications Faculty of Veterinary Medicine
During the course of PARASOL, we produced Ghent University
58 publications in peer-reviewed journals, Salisburylaan 133
74 abstracts in conference proceedings, 9820 Merelbeke
two book chapters and seven PhDs. In addi- BELGIUM
tion, several farming publications and leaf- jozef.vercruysse@ugent.be
lets were produced and talks were given
to farmers and veterinarians in order dis-
seminate the knowledge acquired through
the Parasol-project and promote the TST
approach in the control of nematodes in
ruminants.
Partners
University of Bath Adrian Wolstenholme bssajw@bath.ac.uk
Claverton Down
Bath
BA2 7AY
UNITED KINGDOM
University of Bristol Gerald Coles eric.morgan@bristol.
Langford House ac.uk
Bristol
BS40 5DU
UNITED KINGDOM
Institut national de la recherche Dominique Kerboeuf, kerboeuf@tours.inra.fr
agronomique Jacques Cabaret,
INRA Base Michel Alvinerie
37380 Nouzilly
FRANCE
Stiftung Tierrztliche Hochschule Hannover Georg von Samson- gvsamson@tiho-
Buenteweg 2 Himmelstjerna hannover.de
30559 Hannover
GERMANY
National Veterinary Institute and Swedish Johan Hglund johan.hoglund@sva.se
University of Agricultural Sciences
SE-751 89 Uppsala
SWEDEN

Moredun Research Institute Frank Jackson frank.jackson@more-
Pentlands Science Park dun.ac.uk
Bush Loan
Penicuik
EH26 0PZ
UNITED KINGDOM
Parasitological Institute Marin Vrady varady@saske.sk
Slovak Academy of Sciences
Hlinkova 3
040 01Kosice
SLOVAKIA
University of Pretoria Gareth Bath gareth.bath@up.ac.za
Private Bag X04 Jan van Wyk jan.vanWyk@up.ac.za
0110 Onderstepoort
SOUTH AFRICA
Institut Agronomique et Vtrinaire Hassan II Boumadiane Berrag b.berrag@iav.ac.ma
Avenue Allal El Fassi Rabat Institutes
BP 6202
Rabat
MOROCCO
Aristotelio Panepistimio Thessalonikis Elias Papadopoulos eliaspap@vet.auth.gr
541 24 Thessaloniki
GREECE
Universita degli Studi di Napoli Federico II Giuseppe Cringoli cringoli@unina.it
Via della Veterinaria 1
80137 Napoli NA
ITALY
Innovis Ltd Eurion Thomas ert@innovis.org.uk
Peithyll Centre
Capel Dewi
Aberystwyth
SY23 3HU
UNITED KINGDOM
National Wool Growers Association Leon de Beer nwga@nwga.co.za
41A Pickering Street
Newton Park
6055 Port Elizabeth
SOUTH AFRICA
Svanova Biotech AB Malik Merza Malik.Merza@svanova.
Uppsala Science Park com
SE-751 83 Uppsala
SWEDEN
PlantaMedium GmbH Sandra Schmidt schmidt@plantame-
Everswinkeler Strae 7 dium.de
48231 Warendorf
GERMANY
Fraenz & Jaeger GmbH Thomas Frnz rolf.dietze@fraenz-
Schurzelter Strae 27 jaeger.de
52074 Aachen
GERMANY
[PARAVAC]
Vaccines against helminth

parasites of livestock of
economic and/or public-
health significance
Summary: alternative, greener and more sustainable Acronym:
Livestock production efficiency is impaired approach is to control these infestations by PARAVAC
by worm infections which are ubiquitous in vaccination but, with one exception, there
Project number:
cattle, sheep and goats worldwide. These are no commercial vaccines available for 265862
cause severely debilitating gastro-intes any of these parasites.
EC contribution:
tinal, respiratory or liver disorders, depend-
EUR 8 944 185
ent on the infecting species. Control of
these parasites relies almost exclusively Aim: Duration:
48 months
on the use of drugs, a solution threatened This proposal aims to deliver at least one
by the global emergence of worm strains of these prototype vaccines to the point of Start date:
which are no longer affected by these uptake by the commercial sector or through 1 April 2011
chemicals. An alternative, greener and more government/philanthropic agencies. This Instrument:
sustainable approach is to control these goal will be addressed by: Collaborative project
infestations by vaccination, but, with one
exception, there are no commercial vaccines (i) developing effective native or syn-
available for any of these parasites. Mem- thetic vaccines, the latter using novel,
bers of the present consortium, with partici- molecular expression systems;
pants from the EU and Switzerland, North (ii) d ef ining t he protec tive immune
and South America, and Africa, including responses induced by these vaccines
three SMEs and one major animal health to order to optimise the structure of
company, have collaborated to develop pro- the antigens and the method of their
totype vaccines with the efficacy predicted delivery;
to control several of the most important of (iii) defining vaccine efficacy with trials in
these livestock parasites as well the tape- both housed and grazing livestock;
worm Echinococcus granulosus in dogs, (iv) providing a platform for training and
which can also cause fatal disease in man. knowledge exchange, which includes
participation in training programmes,
short exchanges of staff, workshops,
Problem: and website provision;
As stated above, control of these para- (v) i nteracting closely with computer
sites relies almost exclusively on the use of modellers, the animal health indus-
drugs, a solution threatened by the global try, farmer organisations and other
emergence of worm strains which are no stakeholders to define required vaccine
longer affected by these chemicals. An characteristics;
(vi) k nowledge exchange/dissemination to Project website

policymakers, scientists, government http://paravac.eu
departments and the general public.
Keywords
livestock, sheep, cattle, helminth disease,
Expected results vaccine
At least one vaccine to reach
commercialisation. Coordinator
Prof. Dave Knox
Moredun Research Institute
Potential applications Pentlands Science Park
Any vaccines developed will be applied Bush Loan
for the control of animal health in graz- Penicuik
ing livestock either globally or regionally. EH26 0PZ
In addition, progress in vaccines against UNITED KINGDOM
Haemonchus and fluke will have application dave.knox@moredun.ac.uk
in human medicine for the development of
vaccines against fluke and schistomiasis.
Partners
Department of Virology, Parasi- Edwin Claerebout edwin.claerebout@ugent.be
tology and Immunology
Faculty of Veterinary Medicine
Ghent University
BELGIUM
Unit for Molecular Glycobiology Prof. Dr Nico Nico.callewaert@dmbr.ugent.be
Laboratory for Protein Bio- Callewaert
chemistry and Biomolecular
Engineering
Department of Biochemistry and
Microbiology, UGent
K.L.-Ledeganckstraat 35
9000 Ghent
BELGIUM
Veterinary Microbiology and Grace Mulcahy grace.mulcahy@ucd.ie
Parasitology
University College Dublin
Belfield Campus
Dublin 4
IRELAND
Lyon Ingnierie Projets Prof. Ann-Francoise petavy@univ-lyon1.fr
43 Boulevard du Petavy
11 novembre 1918
LAtrium BP 32009
69616 Villeurbanne
FRANCE
Institute for Parasitology Prof. Thomas thomas.schnieder@tiho-hannover.
University of Veterinary Medicine Schnieder de
Hannover
GERMANY

Leiden University Medical Center Dr Cornellis Hokke c.h.hokke@lumc.nl
Building 1
Postzone L4-Q
PO Box 9600
2300 RC Leiden
NETHERLANDS
University of Glasgow Dr Collette Britton c.britton@vet.gla.ac.uk
Garscube Estate
Bearsden Road
Glasgow
UNITED KINGDOM
School of Veterinary Science Prof. Diana Williams Williadj@liverpool.ac.uk
The University of Liverpool
Parasitology Unit
Crown Street
Liverpool
L69 7ZJ
UNITED KINGDOM
Deparment of Anatomy and Prof. Jos Prez an1pearj@uco.es
Comparative Pathology
University of Crdoba
Campus de Rabanales
Ctra. MadridCdiz km 396
14014 Crdoba
SPAIN
Institute of Parasitology Prof. John P. Dalton john.dalton@mcgill.ca
McGill University
21111 Lakeshore Road
Sainte Anne de Bellevue
Quebec
H9X 3V9
CANADA
Parasitology and Parasitic Dis- Prof. A. Dakkak a.dakkak@iav.ac.ma
eases Unit
OIE Reference Laboratory for
Echinococcosis/Hydatidosis
Institut Agronomique et Vtri-
naire Hassan II
BP 6202 Rabat-Instituts
MOROCCO
Seccin Bioqumica A. Esteves aesteves@fcien.edu.uy
Facultad de Ciencias
Igu 4225
CP 11400 Montevideo
URUGUAY
Parasites Vectors and Vector- Andrea Spickett spickettA@arc.agric.za
Borne Diseases
Onderstepoort Veterinary
Institute
Pretoria
SOUTH AFRICA

Universidade Estadual Paulista Alessandro Amarante amarante@ibb.unesp.br
Department of Parasitology
Julio de Mesquita Filho
Campus de Botucatu
Botucatu
BRAZIL
Isconova AB Gerd Rundstrm gerd.rundstrom@isconova.com
Uppsala Science Park
Dag Hammarskjlds vg 54 A
SE-751 83 Uppsala
SWEDEN
Institute of Parasitology Peter Deplazes, deplazesp@access.uzh.ch
University of Zurich Prof.
Winterthurerstrasse 266a
8057 Zurich
SWITZERLAND
Service do Parasitologie Samia Lahmar drlsamia@yahoo.fr
cole nationale de
mdecine vtrinaire
2020 Sidi Thabet
TUNISIA
Pfizer Limited Fraser Stewart fraser.p.stewart@pfizer.com
Ramsgate Road
Sandwich
CT13 9NJ
UNITED KINGDOM
Laramedia Gerald Cannon ger@laramedia.net
Iveragh Road
Whitehall 98
Dublin 9
IRELAND
Universidad Nacional de Pedro Ortiz ortizpedroluis@yahoo.es
Cajamarca (UNC)
Av. Atahualpa N 1050
Cajamarca
PERU
Triveritas Julian Braidwood julian.braidwood@triveritas.com
Bank Barn
How Mill
Brampton
CA8 9JY
UNITED KINGDOM
Murdoch University Dr Robert J. Dobson R.Dobson@murdoch.edu.au
School of Veterinary and Bio-
medical Sciences
South St
Murdoch
Western Australia 6150
AUSTRALIA
3.3. Tick-borne diseases

[PiroVac]
Improvement of current
and development of new
vaccines for theileriosis and
babesiosis of small ruminants
Summary PiroVac is a collaborative effort among a
Acronym:
PiroVac is a major international research number of established research groups PiroVac
project designed to develop control meas- working on theileriosis and babesiosis. The
Project number:
ures to combat two major tick-borne dis- consortium also encompasses laboratories
245145
eases of small ruminants, namely theile- involved in malaria research in order that
riosis and babesiosis. This EU-funded scientific and technological knowledge in EC contribution:
EUR 3 million
research programme aims at improving that field can be translated into tools and
existing vaccines, designing new vac- reagents for small ruminant piroplasms. Duration:
cines, and capacity-building in partner Industrial expertise in vaccine development 48 months
laboratories both in Europe and in endemic and delivery systems has also been incorp Start date:
areas. orated in order to maximise the potential 1 April 2010
for translational application. Instrument:
Small ruminant piroplasmosis is a major Collaborative project
threat to livestock production in many
areas of the developing world. Theilerosis Problem
and babesiosis, caused by the protozoan Management of ticks and tick-borne dis-
parasites Theileria lestoquardi, T. uilen- eases (TTBDS) is primarily through the
bergi and Babesia ovis, infect sheep and control of the tick vector using acaricides,
goats causing disease, production loss and although this is unsustainable due to
sometimes death. Consequently, these increasing acaricide resistance and food
diseases have a major impact on ani- safety concerns. In some endemic regions,
mal welfare and stock-holder prosperity attenuated live vaccines have been devel-
throughout the world. oped for bovine piroplasmosis (Theileria and
Babesia) however; there has been limited
By developing effective measures to control development of vaccines for small ruminant
these serious diseases, the PiroVac project piroplasmosis.
represents a major contribution to achiev-
ing the United Nations millennium devel-
opment goals of food security, food safety, Aim
poverty alleviation, animal welfare and The PiroVac project was developed as
environmental sustainability. an integrated approach, encompassing
immunology, molecular biology, bioinformat- molecules associated with attenuation of

ics and genetic engineering, together with parasite virulence to be included in the devel-
pathogen genomics and host genetics, and opment of safe and efficacious live vaccines.
is directed at addressing two broad aims: For the improvement of the attenuated T.
lestoquardi vaccine, a combination of the
(i) development of effective and reli- existing vaccine with subunit vaccines will be
able vaccines for use in disease control examined for synergistic effects. The specific
campaigns for sustainable livestock goals of the project are:
development;
(ii) c apacity-building for the sustainable (a) i mprovement and development of
implementation of integrated control live attenuated vaccines for the con-
measures required for disease control trol of small ruminant theileriosis and
and/or eradication through increas- babesiosis through determining the
ing scientific knowledge, training and effectiveness of attenuation, using:
improvement of infrastructure. (i) i n vivo assessment of attenu-
at ion, a nalysing clinic al a nd
immu nologic al cr it er ia (bot h
humoral and cellular responses) of
immunised and challenged animals;
(ii) subtractive libraries and microarray
analysis for the identification of
attenuation markers;
(b) subunit vaccine design through:
(i) identification of suitable antigens
using a combination of genomics,
bioinformatics and gene expression
analysis coupled with experimental
confirmation of antigen localisation
and presentation to facilitate
antigen discovery, parasite mol-
ecules involved in host cell invasion,
activation of cytokine-producing
CD4+ T cells and NK cells and acti-
vation of cytotoxic T-lymphoc ytes
involved in killing of T. lestoquardi-
PiroVac will exploit technology to improve infected leucocytes will be identified;
the existing T. lestoquardi vaccine and design (ii) immunological characterisation of
new vaccines focusing on malignant theile- the identified antigens as potential
riosis and babesiosis in small rumina nts. A vaccine candidates.
wealth of scientific information will be gener- (c) vaccination trials using:
ated that will facilitate the upgrading of pro- (i) live attenuated vaccines;
duction systems using more productive, but (i) recombinant protein and DNA
disease-susceptible, breeds for improving the vaccines.
genetics of local flocks. The overall objective
of the project is to ensure food security and
to improve food safety by improving control Expected results
measures for small ruminant piroplasmosis The project will contribute to an under-
caused by T. lestoquardi, T. uilenbergi and standing of the immunological and molecu-
B. ovis. To achieve these goals, the project lar mechanisms involved in host-pathogen
will assess parasite diversity and identify interaction. As important by-products of
the project, reagents required for the char- for the commercial exploitation of the
acterisation of the innate and adaptive products).
immunity of small ruminants will be gener-
ated together with the genome sequences Project website
of the three pathogens under study. http://www.theileria.org/pirovac/
The PiroVac project is based on the convic- Keywords

tion that the interface between genomics, small ruminants, piroplasmosis, Theileria
immunology and vaccinology offers the lestoquardi, Theileria uilenbergi, Babesia
best prospect for major breakthroughs in ovis, vaccine
vaccine discovery and development. Over-
all, vaccine development will contribute to Logo
reducing losses in animal production due
to piroplasmosis of small ruminants and
improve the life quality of both farmers and
consumers.
Generation of knowledge and tools
required for vaccine production.
Vaccines useful for upgrading schemes
using more susceptible but productive
breeds.
Progress into policy innovations and
strategies that will meet critical millen- Coordinator
nium development goals: food security, Jabbar Ahmed
food safety, poverty alleviation (e.g. Research Center Borstel
improvement of small farmer income), Parkallee 22
animal welfare and environmental 23845 Borstel
sustainability. GERMANY
Exploitation of the data for commer- Tel. +49 4537188-4280
cial purposes by collaborating with Fax +49 4537188-6270
the industry (arrangements have been jahmed@fz-borstel.de
made with the industrial participant
Partners
University of Glasgow, Prof. Andy Tait a.tait@vet.gla.ac.uk
United Kingdom (UGLA) Prof. Brian Shiels b.shiels@vet.gla.ac.uk
and willie.weir@glasgow.ac.uk
Dr William Weir
Lanzhou Veterinary Prof. Yin Hong yinhong@public.lz.gs.cn
Research Institute, China
(LVRI)
Bernhard Nocht Institut fur Dr Volker Heussler volker.heussler@izb.unibe.ch
Tropenmedizin, Germany
(BNITM)

Institute national de la Prof. Gordon Langsley gordon.langsley@inserm.fr
sant et de la recherche
mdicale, France (Inserm)
Ministry of Agriculture and Dr Varda Shkap shkapv@int.gov.il
Rural Development, The
Kimron Veterinary Institute,
Israel (KVI)
Universitaet Bern, Prof. Dirk Dobbelaere dirk.dobbelaere@mopa.unibe.ch
Switzerland (UNIBE)
The Royal Veterinary Col- Prof. Declan McKeever dmckeever@rvc.ac.uk
lege, United Kingdom (RVC)
Adnan Menderes Dr Tulin Karagenc tulinkaragenc@yahoo.com
University, Turkey
(ADU)
The University of Edinburgh, Prof. Ivan Morrison Ivan.Morrison@ed.ac.uk
United Kingdom (UE) and Liz.Glass@roslin.ed.ac.uk
Prof. Liz Glass
Parco Tecnologico Padano Dr John Williams john.williams@tecnoparco.org
s.r.l., ltaly (PTP)
Instituto de Biologia Experi- Dr Abel Martn oliva@itqb.unl.pt
mental e Tecnologica, Gonzlez Oliva
Portugal (IBET) and
Prof. Virglio Estlio do
Rosrio
cole nationale vtrinaire Prof. Alain Chauvin alain.chauvin@vet-nantes.fr
de Nantes, France (ENVN)
Centro de Investigaciones Dr Monica mflorin@cnia.inta.gov.ar
en Ciencias Veterinarias Florin-Christensen leoschnittger@googlemail.com
y Agronomicas, Argentina and
(CICVyA) Dr Leonhard Schnittger
Intervet International BV, Prof. Theo Schetters theo.schetters@sp.intervet.com
Netherlands
3.4. Porcine circovirus diseases

[PCVD]
Control of porcine
circovirus diseases (PCVDs):
towards improved food
quality and safety
Summary PCVD that are being applied across all
Acronym:
This PCVD project was initiated on 1 EU Member States and third countries; PCVD
December 2004. It was a 51-month, 2. a reduction in the load of secondary
Project number:
research-based FP6-funded project with bacterial infections in pig herds, accom-
513928
the specific objectives of providing sound panied by a consequential reduction in
scientific information which could be the use of antibiotic therapy and risk of EC contribution:
EUR 3 450 000
used to promote food quality and safety acquired antibiotic resistance;
through the control of porcine circovirus 3. an increase in the quality and safety of Duration:
diseases (PCVDs) within EU Member States food products derived from pigs; 51 months
and regions. The project brought together 4. establishment of a common standard- Start date:
a multidisciplinary scientific team of 15 ised and harmonised reagent bank for 1 December 2004
partners representing diagnostic insti- the diagnosis and study of PCVD; Instrument:
tutes, universities and industry. This project 5. determination of the molecular mech Specific Targeted
established an EU-led multidisciplinary con- anisms of PCV2 replication and patho- Research or
sortium containing expertise in epidemiol- genesis and the early replication sites Innovation Project
(STREP)
ogy, pig genetics, pig nutrition, pathology, of PCV2 in pigs;
molecular biology, immunology, vaccinol- 7. elucidation of the role of nutrition and
ogy, bacteriology and PCV virology to gen- other environmental factors in the full
erate scientifically sound information on the clinical expression of PCVD;
aetiology and early pathogenesis of PCVDs. 8. elucidation of the early interactions of
The project met all the objectives and the PCV2 with the porcine immune system
information generated has been used to relevant to susceptibility or resistance
generate control measures for PMWS/PCVD, to PCVDs.
resulting in the reduction of the use of anti-
biotics and of secondary zoonotic bacterial
infections, meeting consumer concerns for Problem
quality/safety of pork products. The results In 1997, the first reports of a mystery wast-
can be summarised as follows: ing disease in pigs in Canada and France
started to appear in conference proceed-
1. successful development of effective and ings and the farming press and eventually
consistent control measures for PMWS/ in peer-reviewed journals. Following these
preliminary reports the disease, named post- 3. to identify the common co-factors/trig-
weaning multisystemic wasting syndrome gers in epizootic PCVD scenarios neces-
(PMWS), quickly spread around the world as sary for the full development of clinical
a global epizootic causing severe economic disease;
losses to pig producers and restricting the 4. to determine the role of nutrition in the
movement of pigs and pig products. EU susceptibility/resistance to PCVD;
researchers working within projects funded 5. to determine the sites of replication of
under the fifth and sixth framework pro- PCV2 and early pathogenesis of PCVDs;
grammes (FP5 and FP6), in collaboration 6. to elucidate the early interactions
with industry and colleagues from North of PCV2 virus with the host immune
America, have been at the forefront of stud- system;
ies on PMWS and porcine circoviruses that 7. to elucidate the role of porcine genetics
have defined the disease and helped develop in susceptibility/resistance to PCVD;
commercial diagnostics and vaccines. At the 8. to determine the molecular processes
initiation of this FP6 project in 2004, PMWS of PCV2 replication (18 months) and
was the most serious global disease affect- virulence;
ing the swine industry. Surveys at the time 9. to standardise and harmonise and
showed that the prevalence of PMWS and distribute reagents, and SOPs for use
PCVD were high in all parts of the world and within the consortium.
had both a strong sanitary and economical
impact. Wasting disease was the predomi-
nant clinical sign, and mortality, other infec- Results
tions and their direct consequences and the T his projec t has been successfully
use of antibiotics were universally increased. completed and all objectives were met.
At the beginning of the outbreaks, mortal-
ity rates peaked at 3540 %, sometimes A classical commercial vaccine was suc-
higher in some batches, and mainly occurred cessfully tested in trials in Canada, Den-
between 6 and 14 weeks of age. In 2004, no mark and Sweden and successful trials on
commercial vaccine products were available disease prevention using combined vaccine/
for control of PMWS. nutritional intervention were completed.
Vaccination is now a major tool to prevent
disease, good nutrition is very efficacious
Aim and vaccination and good nutrition are syn-
The strategic objective of this project ergistic. In the hands of the consortium,
was to generate information and control more advanced experimental vaccines did
measures for porcine circovirus diseases not appear superior to a classical marketed
that would have a positive impact on the product. Based on the data generated, it
health and welfare of pigs and meet con- can be concluded that piglet vaccination
sumer concerns for quality and safety of prevents PMWS and improves the growth
pork products. This overarching objective performance of the piglets but was lim-
was achieved by completing the nine minor ited by maternal immunity. Sow vaccin
objectives outlined below: ation improves reproduction parameters,
prevents PMWS mortality and decreases
1. to apply the information generated to global mortality and improves growth and
the elimination and/or control of PCVD; performance of the piglets and pigs.
2. to initiate and maintain a proactive
information dissemination programme Within the project, there was an active
aimed at all relevant stakeholders, programme of knowledge transfer and
including consumers, producers and interaction with stakeholders throughout
policymakers; the EU and internationally. The aim of the
knowledge transfer programme was to infection and also during clinical manifest
ensure good communication of the results ation, which enables future bio-informatic
of the programme at both the scientific analysis. Based on these studies, the early
level and at a less technical level to pro- interactions of PCV2 with the immune sys-
ducers, veterinary surgeons and general tem have been elucidated. The mode of
stakeholders so that relevant results could PCV2 interacting with different cell subsets
be applied as quickly as possible to improve as well as with production of antibodies
the health and welfare of pigs. Close col- and cytokines has been described in detail
laboration with the sixth framework specific in scientific publications.
support action PCVD-SSA helped to transfer
both knowledge and skill sets to new Mem- PCV2 is a small virus with a simple genomic
ber States, accession countries and beyond. organisation (expressing only few proteins),
The project website was central to the which shows a high similarity to PCV1. At
knowledge transfer effort and it is intended the initiation of this project, we did not have
to maintain this as a source of information an understanding, at the molecular and cel-
from the project beyond the lifetime of the lular level, of how infection of swine with
project so that the knowledge transfer will PCV2 caused a complex and severe disease
be ongoing. such as PMWS. By investigation of the entry
and the replication of the virus in different
Within the project a complementary feed- cell lines, highly deviating types of infec-
stuff comprising short-, medium- and long- tion were found. We have focused on the
chain fatty acids was shown to improve molecular interaction of PCV1 and PCV2
growth rates and FCR in field trials on with cellular components and the identi-
units with pigs diagnosed with PCVD. Pigs fication of up- or down-regulated genes
offered the feedstuff had lower levels of after PCV2 infection. This process has led
PCV2 excretion than pigs not offered the to the identification of several highly inter-
complementary feedstuff. The complemen- esting genes that may be involved in PCV2-
tary feedstuff did not work by improving induced pathogenesis.
digestibility but by improving utilisation of
the feed. At the initiation of this project, consortium
members were using a wide range of dif-
It was also shown within the project that ferent reagents and biologicals to diagnose
porcine embryos and foetuses are suscep- PCVD and carry out experimental stud-
tible to PCV2 infection and that infection ies. A work package (WP) was designed to
leads to foetal mortality irrespective of standardise and harmonise these reagents
genotype or clinical background of the virus across the consortium in an attempt to
strain. Sequential studies have elucidated ensure that results generated by consor-
that mucosal tissues are sites of early rep- tium members were comparable. In addi-
lication and these findings have been cor- tion, the WP distributed SOPs, reagents
roborated by evidence of replicative forms and biologicals to researchers and diag-
of virus in epithelial and endothelial cells nosticians outside the consortium in an
in clinical cases of PCV2 infection. Effects attempt to standardise diagnostics and
of PCV2 on intestine and intestinal cells research findings across EU Member States
have been particularly addressed. Altered and ACCs. Reagents and procedures were
transcriptome patterns after PCV2 infec- standardised and harmonised by ring test-
tion have been shown in vitro and in vivo ing and distributed to consortium members
and are suggestive for functional changes and a large number of laboratories out-
in these cells. By microarray analysis, infor- side the consortium. In addition, a number
mation has been generated of differentially of SOPs were written and posted on the
regulated gene expression early after PCV2 PCVD webside and a number of consortium
recommendations for diagnostic proce- have enabled successful technology trans-

dures were also posted and presented at fer and training of young scientists within
international meetings. EU Member States and third countries.
Throughout the lifetime of the consortium, Results generated from this project
we have focused on the characterisation on control measures and vaccinations
of new PCV2 isolates and new biologicals against PCV2 have therefore contributed
produced by consortium members. To this significantly to:
end, the consortium was the first group to
recognise and characterise new PCV2 geno- improved welfare for swine;
types (partners 1 and 10) recovered from protection of the environment;
pigs in Sweden. This was quickly followed protection of the customer, with much
by the characterisation of new genotypes less antibiotic use;
from pigs in Denmark and Spain. The asso- very significant economical benefits to
ciation of genotypes of PCV2 with virulence farmers.
and disease progression is still unclear and
a matter of debate. The strategic objective of the consortium
was fully achieved by generating infor-
mation and control measures for porcine
Potential applications circovirus diseases that will have a positive
This collaborative project successfully impact on the health and welfare of pigs
brought together scientists in 15 institutes and meet consumer concerns for quality
from 10 countries to apply a multidiscip and safety of pork products.
linary approach to the understanding
and eventual control of an economically
important disease of swine. References/publications
Misinzo, G., Delputte, P.L., Meerts, P.,
The project has been successfully com- Lefebvre, D.J., Nauwynck, H.J. (2006),
pleted within the agreed budget, with the Porcine circovirus 2 uses heparan sulfate
vast majority of the anticipated milestones and chondroitin sulfate B glycosaminogly-
and deliverables being met. The project has cans as receptors for its attachment to
significantly contributed to the understand- host cells, Journal of Virology, 80, 2006,
ing of porcine circovirus diseases (PCVDs), 34873494.
with 53 publications in the peer-reviewed
literature, and maintained and expanded Steinfeldt, T., Finsterbusch, T., Mankertz,
the established internationally recognised A. (2006), Demonstration of nicking/join-
expertise in these diseases in Europe. ing activity at the origin of DNA replication
Importantly the project has directly con- associated with the Rep And Rep proteins
tributed to the control of these diseases of porcine circovirus type 1, Journal of
around the world by application in the field Virology, 80, 2006, 62256234.
of results generated within the project on
vaccines, epidemiology and nutritional Hasslung Wikstrm, F., Meehan, M.B., Berg,
intervention. The practical application of M., Timmusk, S., Elving, J., Fuxler, L., Mag-
results generated from studies associ- nusson, M., Allan, M.G., McNeilly, F., Fossum,
ated with this project has also resulted in C. (2007), Structure-dependent modula-
improved diagnostic procedures for PCVDs tion of alpha interferon production by por-
being applied around the world. cine circovirus 2 oligodeoxyribonucleotide
and CpG DNAs in porcine peripheral blood
The association and interactions of the mononuclear cells, Journal of Virology, 81,
PCVD STREP with the NMSACC-PCVD SSA 2007, 49194927.
Mateusen, B., Maes, D., Van Soom, A., Project website

Lefebvre, D., Nauwynck, H.J. (2007), http://www.pcvd.org
Effect of a porcine circovirus type 2 infec-
tion on embryos during early pregnancy, Keywords
Theriogenology 68, 2007, 896901. porcine circovirus, virology, swine, wast-
ing disease, postweaning multisystemic
Segals, J., Olvera, A., Grau-Roma, L., Char- wasting syndrome
reyre, C., Nauwynck, H., Larsen. L., Dupont.
K., McCullough, K., Ellis, J., Krakowka, S., Coordinator
Mankertz, A., Fredholm, M., Fossum, C., Tim- Prof. Gordon Allan
musk, S., Stockhofe-Zurwieden, N., Beattie, Department of Veterinary Science
V., Armstrong, D., Grassland, B., Baekbo, P., Queens University of Belfast
Allan, G. (2008), PCV-2 genotype definition Stoney Road
and nomenclature, Veterinary Record, 162, Belfast
2008, 867868. BT4 3SD
UNITED KINGDOM
Gordon.allan@afbini.gov.uk
Partners
MERIAL SAS Dr Catherine Charreyre catherine.charreyre@merial.com
29 Avenue Tony Garnier
69007 Lyon
FRANCE
Faculty of Veterinary Medicine Prof. Hans Nauwynck hans.nauwynck@ugent.be
Laboratory of Virology
Ghent University
Salisburylaan 133
9820 Merelbeke
BELGIUM
Danish Institute for Food and Dr Lars Erik Larsen lel@dfvf.dk
Veterinary Research
Blowsvej 27
1790 Copenhagen V
DENMARK
Dept of Immunology Dr Ken McCullough kenneth.mccullough@ivi.admin.ch
Institut fuer Viruskrankheiten
and Immunprophylaxe
Sensemattstrasse 293
3147 Mittlehausen
SWITZERLAND
Department of Veterinary Dr John Ellis ellisj@duke.usask.ca
Microbiology
Western College of Veterinary
Medicine University of
Saskatchewan
52 Campus Drive
Saskatoon 57N 5B4
CANADA

The Ohio State University Dr Steven Krakowka krakowka.1@osu.edu
Department of Veterinary
Biosciences
College of Veterinary Medicine
1925 Coffey Road
Columbus OH 43210
UNITED STATES OF AMERICA
P11 Neuartige Viren / Dr Annette Mankertz A.Mankertz@rki.de
Xenotransplantation
Robert Koch-Institut
Nordufer 20
13353 Berlin-Wedding
GERMANY
The Royal Veterinary and Prof. Merete Fredholm mf@kvl.dk
Agricultural University
Department of Animal Science
and Animal Health
Division of Animal Genetics
Groennegaardsvej 3
1870 Frederiksberg C
DENMARK
Department of Molecular Prof. Caroline Fossum Caroline.Fossum@vmm.slu.se
Biosciences
Section for Veterinary Immu-
nology and Virology
Swedish University of Agricul-
tural Sciences
Biomedical Centre PO Box 588
SE-751 23 Uppsala
SWEDEN
Animal Sciences Group Dr Norbert Stockhofe Gerard.Wellenberg@wur.nl
Wageningen UR
Infectious Diseases
Edelhertweg 15
PO Box 65
8200 AB Lelystad
NETHERLANDS
Devenish Nutrition Ltd Dr Violet Beattie violet.beattie@devenishnutrition.
96 Duncrue Street com
Belfast
BT3 9AR
UNITED KINGDOM
Centre de Recerca en Sanitat Prof. Joaquim Segals Joaquim.Segales@uab.es
Animal
Facultat de Veterinaria
Universitat Autnoma de
Barcelona
08193 Bellaterra
Barcelona
SPAIN

Veterinary Programme Mr Derek Armstrong, derek.armstrong@bpex.org.uk
Manager MVB, MRCVS
BPEX
Veterinary Department
Stoneleigh Park
Kenilworth
Warwickshire
CV8 2TL
UNITED KINGDOM
Chef de lUnit Gntique Dr Andr Jestin a.jestin@ploufragan.afssa.fr
virale et Bioscurit (UGVB)
Agence franaise de scu-
rit sanitaire des aliments
(AFSSA)
22440 Ploufragan
FRANCE
The National Committee for Dr Poul Baekbo
Pig Production
Danish Bacon andd Meat
Council
Section for Veterinary
Research and Development
Vinkelvej 11
8620 Kjellerup
DENMARK
[NMSACC-PCVD]
PCVD: Towards improved

food quality and safety
within EU new Member States
and candidate countries
Summary control of PCVD were harmonised in a
Acronym:
NMSACC-PCVD
The NMSACC-PCVD specific support action range of NMSs and ACCs, and information
(SSA) was initiated on 1 January 2006. dissemination across NMSs and ACCs was
Project number:
This SSA was designed to support infor- achieved through a series of five work-
518432
mation dissemination to, and training of, shops (two additional workshops over and
EC contribution: young scientists and various groups of above those outlined in the original techni-
EUR 397 340
stakeholders and end-users from new cal annex). A network of training opportu-
Duration: Member States (NMSs) and acceding and nities for young scientists across EC Mem-
48 months candidate countries (ACCs) in a wide range ber States and ACCs has been established.
Start date: of aspects related to porcine circovirus
1 January 2006 diseases that were being explored and
Instrument: researched within the PCVD STREP. This The results can be summarised
specific support SSA allowed scientist and other end-users as follows
action and stakeholders in NMSs and ACCs to 1. Reagents and protocols for the study
benefit from the consolidation and expan- of PCVD have been harmonised across
sion of reagents, information and training a number of NMSs and ACCs through
opportunities on PCVD and other pig dis- a series of ring tests. Reagents have
eases important in food safety. The SSA been distributed to 21 institutes in 15
specifically addressed the linking of NMSs countries.
and ACCs to the PCVD STREP, resulting 2. A series of five workshops, successfully
in a multidisciplinary network of over 37 completed in NMSs, incorporated dele
partners. The harmonisation of reagents, gates from diagnostic and research lab-
dissemination of information and training oratories, field veterinarians and swine
of young researchers in a network linked producers. A workshop in Russia was
to an already established PCVD project of included in this project.
international standing consolidated and 3. An information dissemination network
strengthened the globally acknowledged was established that included a spe-
centre of expertise on PCVD within the cific website. Funding was made avail-
EU. The interactions established within able for 16 young scientists to attend
this SSA with colleagues within the NMSs international meetings.
and ACCs assisted preparations for future 4. A successful extensive training and
Community research activities. The pro- exchange programme for young scien-
ject met, and exceeded, all the objectives. tists from NMSs and ACCs and the Rus-
Reagents and protocols for diagnosis and sian Federation was completed. In this
programme, 32 young scientists from young researchers, dissemination of infor-

11 countries received training in STREP mation to end-users and assisting NMSs
laboratories. and ACCs with preparations for future Com-
5. Young scientists from NMSs and ACCs munity research activities. The objectives of
have been trained and empowered in a the SSA were:
number of aspects of organisation and
management of an EC project. 1. to harmonise across NMSs and ACCs the
reagents and procedures;
2. to establish by means of a programme
Problem of three joint workshops a common
In 1997, the first reports of a mystery understanding of the state-of-the-art
wasting disease in pigs in Canada and knowledge of the pathogenesis, pre-
France started to appear in conference vention and control of PCVDs; these
proceedings and the farming press and workshops were to include participa-
eventually in peer-reviewed journals. Fol- tion by end-users, industry and young
lowing these preliminary reports the dis- researchers;
ease, named postweaning multisystemic 3. to disseminate information and know
wasting syndrome (PMWS), quickly spread ledge on PCVD across NMSs and ACCs;
around the world as a global epizootic caus- 4. to develop a scientist exchange pro-
ing severe economic losses to pig produc- gramme between NMSs/ACCs and PCVD
ers and restricting the movement of pigs STREP members, including former USSR
and pig products. At the initiation of the countries;
FP6 STREP on PMWS and PCVD (Control of 5. to complete and distribute a com-
PCVD: Towards improved food quality and prehensive infor mation dissemi-
safety: FOOD-CT-2004-513928) in 2004, nation package on outcomes and
PMWS/PCVD was the most serious global achievements of PCVD SSA/STREP.
disease affecting the swine industry. Sur-
veys at the time showed that the preva-
lence of PMWS and PCVD were high in all Results
parts of the world, including NMSs and This project has been successfully com-
ACCs, and had both a strong sanitary and pleted and all objectives were met. Some
economical impact. It was also clear from objectives were exceeded.
these surveys that there was an urgent
need for information dissemination and Harmonisation of reagents
training platforms for scientists, field vet- At the initiation of the SSA, work within
erinarians and swine producers in NMSs the PCVD STREP had already begun on
and ACCS be set up to facilitate trans- the harmonisation of reagents and pro-
fer of knowledge gained within the PCVD cedures within laboratories in Western
STREP and to help combat this devastating Europe and Canada. From 2006, this work
disease. was expanded within the SSA, and NMSs
and ACCs were included. Partner 5 (NVRI,
National Veterinary Research Institute,
Aim Poland) was responsible for further har-
The strategic objective of the NMDACC- monisation studies, ring trials within SSA-
PCVD SSA was to establish an EU network affiliated institutes and the circulation of
of researchers and end-users that linked standardised reagents and protocols to
21 institutes and stakeholders in NMSs NMSs and ACCs. At the termination of the
and ACCs to an established EU STREP on SSA, 21 institutes in 15 countries in NMSs
the control of PCVD. The SSA focused on and ACCs had been supplied with reagents
the harmonisation of reagents, training of and procedures.
Workshops themselves of this oppor tunity, and

At the initiation of this project, three work- attended international meetings around
shops were envisaged within the duration the world.
of the three-year project. As a result of sav-
ings made within the initial budget, a total Scientific exchange programme
of five workshops and a meeting for young At the heart of this SSA was the develop-
PhD students were successfully completed. ment, training and social interaction of
These workshops were all held in NMSs or young scientists from across EU Member
ACCs countries, with the exception of Work- States and ACCs. Throughout the lifetime
shop 5 which was held in Russia. In all of of the project, special attention was given
these workshops, scientists were encour- to trying to ensure that these young sci-
aged to interact with field veterinarians entists were engaged and developed at
and swine producers to facilitate two-way all levels of the project. WP4 focused on
communication between the field and an exchange programme for young scien-
the laboratory. Written feedback from the tists from NMSs and ACCs to visit, interact
workshops supplied by delegates has indi- with and learn from their colleagues in the
cated that this format was very successful, STREP institutes. Reciprocal visits from
with an average approval rating of over STREP institutes to NMSs and ACCs were
90% for each workshop. also arranged.
Information dissemination A total of 32 young scientists (in excess

Knowledge dissemination was achieved at of the original target of 20) from 15 east
all levels within this project and was coord European countries participated in this pro-
inated by Partner 4 (Veterinary Research gramme. The success of this SSA can, in
Institute, Czech Republic). A dedicated web- part, be measured by the enthusiastic par-
site was set up within the Veterinary Bio- ticipation of young scientists from across EC
technology, Epidemiology and Food Safety Member States and ACCs in all aspects of
Network CENTAUR (http://centaur.vri.cz) the programme and their constructive inter-
providing access to PCVD news, training actions with one another, both scientific and
courses in the EU and third countries, a social, across cultural and political divides.
handbook on EU funds 200713, workshops
and translated versions of the STREP news- Management of the SSA
letter for downloading in Croatian, Czech, The SSA was successfully managed by the
English, Hungarian, Lithuanian, Macedonian, coordinator and the project management
Polish, Romanian and Slovak. The website team. This can be seen by the savings that
also provided a database of new papers were made within the original costings of the
published with abstracts (37 in 2007; 526 project and the additional training of young
since 1985). The database was updated scientists through two extra workshops and
every time a new publication appeared on 12 extra technology transfer visits that was
the Web of Science. The records (authors, achieved without additional cost to the EC.
title and source) are available on the
web page, listed according to the authors or
the year of publication. Importantly, within Potential applications
this work package, a funded programme The SSA linked to the PCVD STREP has
was available to help young researchers focused on information dissemination, the
attend international meetings to present development of young scientists in NMSs
results on their PCVD-related research. and ACCs and the interaction of laboratory-
based research with end-users in the field.
Within the lifetime of the SSA, 16 young The SSA was expanded beyond its original
scientists from 10 countries availed conception to include affiliates in Russia,
Ukraine, Serbia and Macedonia. In addition, very significant economical benefits to

extra outputs were achieved over and above farmers;
those outlined in the technical annex, and a platform of young scientists across EU
included two additional workshops and more Member States trained in PCV2-related
training exchange visits for young scientists. matters.
The association and interactions of the

PCVD STREP with the NMSACC-PCVD SSA Keywords
have enabled successful technology trans- porcine circovirus, virology, swine, wast-
fer and training of young scientists within ing disease, postweaning multisystemic
EU Member States and third countries. wasting syndrome, new Member States
Results generated from this project on Coordinator

training and information dissemination on Prof. Gordon Allan
PMWS and PCVD have contributed signifi- The Queens University of Belfast
cantly to the following outcomes in NMSs Department of Veterinary Science
and ACCs: Stoney Road
Belfast
improved welfare for swine; BT4 3SD
protection of the environment; UNITED KINGDOM
protection of the customer by the much Gordon.allan@afbini.gov.uk
decreased use of antibiotics;
Partners
ORGANISATION CONTACT CONTACT
MERIAL SAS Dr Catherine Charreyre catherine.charreyre@merial.com
29 Avenue Tony Garnier
69007 Lyon
FRANCE
BPEX Mr Derek Armstrong, MVB, derek.armstrong@bpex.org.uk
Veterinary Department MRCVS
Stoneleigh Park Veterinary Programme
Kenilworth Manager
Warwickshire
CV8 2TL
UNITED KINGDOM
Veterinary Research Prof. Karel Hruska hruska@vri.cz
Institute
Hudcova 70
621 32 Brno
CZECH REPUBLIC
National Veterinary Dr Tomasz Stadejek, DVM, stadejek@piwet.pulawy.pl
Research Institute PhD, DSc
Department of Swine
Diseases,
Partyzantow 57
14-100 Pulawy
POLAND
ORGANISATION CONTACT CONTACT

University of Veterinary Prof. Stefan Vilcek, Ing., vilcek@uvm.sk
Medicine PhD, DSc
Department of Infectious
Diseases and Parasitology
Komenskeho 73
041 81 Kosice
SLOVAKIA
Szent Istvn University Dr Tams Tuboly, DVM, PhD tuboly.tamas@aotk.szie.hu
Faculty of Veterinary
Science
Department of Microbiology
and Infectious Diseases
Budapest
Hungria krt. 2325
1143
HUNGARY
Croatian Veterinary Institute Dr Lorena Jemeri, PhD Dr. jemersic@veinst.hr
Virology Department vet. med.
Savska cesta 143
10 000 Zagreb
CROATIA
3.5. Porcine reproductive

and respiratory syndrome
[PoRRSCon]
New tools and approaches

to control porcine reproductive
and respiratory syndrome
in the EU and Asia
development. Two of these partners are
Summary leading European pharmaceutical compa- Acronym:
PoRRSCon
Porcine reproductive and respiratory syn- nies that will guide the consortium in the
drome virus (PRRSV) is the major cause direction of exploitable results. By joining Project number:
245141
of reproductive and respiratory problems their strengths, they have an ideal posi-
in pigs worldwide. It is endemic in Europe tion to be successful in one of the most EC contribution:
and other parts of the world and in the difficult challenges in pig health: control- EUR 2 999 070
United States, it is estimated that losses ling PRRS. Duration:
due to PRRSV exceed USD 560 million 54 months
annually. High abortion rates, mortality of Start date:
pre-weaned piglets and respiratory dis- Problem 1 January 2010
ease in fattening pigs are the main fea- PRRSV is a positive-sense, single-stranded
Starting date:
tures of this syndrome. Controlling this RNA enveloped virus classified within the collaborative project
disease is a top priority in pig producing genus Arterivirus. Nowadays, two geno-
countries. Due to mutations at a high fre- types are recognised (1 and 2) that origin
quency, new variants of the virus appear ally were described as European and
that are no longer effectively controlled American because of the geographic ori-
by the commercial vaccines. In addition, gin of their prototypic strains. The Euro-
highly virulent variants emerge, leading to pean genotype is further separated in
high losses. With regard to animal welfare three subtypes (I, II, III). It has been clearly
and agricultural economics, there is an shown that the best protection is induced
urgent need to control PRRS. Furthermore, when the vaccine virus belongs to the
the abusive use of antibiotics to control same genotype. Due to its high genetic
PRRSV-associated respiratory problems variability, highly pathogenic strains of
may lead to resistance that may endanger this RNA virus have emerged, causing
public health. PoRRSCon is an initiative of up to 2030 % mortality (such as High
14 partners originating from Europe and Fever Disease in China, atypical PRRS in
Asia with strong expertises in virology America and disease caused by subtype III
and immunology. They are doing front- European strains). These new strains are
line research on PRRSV and/or vaccine difficult to control by currently available
vaccines. PRRS is also undermining the Expected results

pigs immunity, leading to extensive bac- To control PRRS, a better understanding
terial infections in PRRSV infected pigs. of the pathogenesis of PRRSV infections is
Antibiotics are consequently intensively essential. With divergent strains emerging
used in order to control these bacterial in Europe and Asia, the characterisation of
infections, leading to antibiotic resistance. the different PRRSV isolates across the two
The only sustainable defence will be the continents is pivotal, with work being carried
development of adaptable, inactivated, out by partners in China, Denmark, Poland
vector and attenuated marker vaccines, and Vietnam. To understand the complex
which can safeguard the pig industry and mechanism by which the disease is caused,
animal welfare for the future. partners in Belgium, the Netherlands and
the United Kingdom are comparing the
pathogenesis of low virulence strains with
Aim highly virulent or pathogenic ones. Concom-
The aim of this project is to develop itantly, partners in Spain, the Netherlands
new tools and approaches to control and the United Kingdom are comparing the
PRRS inthe EU and Asia. To reach this immunological parameters of these differ-
final goal,the following objectives are ent strains during infection: the complex
forwarded: factors which cause immune function vari-
ation. At the fundamental level, work defin-
characterise genetically and anti ing the genetic base of PRRSV virulence is
genically current PRRSV isolates in being carried out by partners in China, Bel-
Europe and Asia; gium and Italy, who are seeking to identify
have a better understanding of the com- the cellular genes associated with virulence/
plex pathogenesis of PRRSV infections, pathogenicity. The Belgian partner is also
immune response against PRRSV and studying the entry of high and low virulence
immune modulation by PRRSV; strains in its target cell, the macrophage,
define the genetic base of PRRSV while partners in Belgium, Spain, the Neth-
virulence; erlands and the United Kingdom are analys-
identify PRRSV proteins and domains on ing the immunobiology of the virus.
these viral proteins that are involved in
the induction of the immunity against The development of a new generation of
PRRSV and in the immune modulation adaptable inactivated, vector and attenu-
of PRRSV; ated marker vaccines which can be adapted
develop new generation, efficacious for temporary changes and geographical
and safe marker vaccines that can be differences is being carried out by partners
adapted to temporary changes and across China, Belgium, Spain and Switzer-
geographical differences; land. Concomitantly, the development of
develop DIVA assays that allow differ- DIVA serological assays, which will allow dif-
entiation of infected from vaccinated ferentiation between infected and vaccinated
animals. pigs, is being carried out by one of our Span-
ish partners (the biotech company Ingenasa).
Ultimately, it will be possible to set up a The PoRRSCon consortium consists of Euro-
control strategy by combining marker vac- pean and Asian PRRS experts who are in con-
cines with DIVA (differentiation of infected tact with national and international authori-
from vaccinated animals) assays. The tools ties and farmer organisations. Furthermore,
and approaches developed in PoRRSCon will the consortium has very good interactions
be made available to all national and inter- with PRRS experts in the United States. The
national authorities that want to solve the final goal is to coordinate PRRS control on
PRRS problems anational, European and world level.
Potential applications Vanhee, M., Van Breedam, W., Costers,

Adaptable inactivated, vector and attenu- S., Geldhof, M., Noppe, Y., Nauwynck, H.
ated marker vaccines for PRRSV, DIVA (2011), Characterisation of antigenic
serological assays for PRRSV regions in the porcine reproductive and
respiratory syndrome virus by the use of
peptide-specific serum antibodies, Vaccine,
References/publications 29(2930):47944804.
Van Breedam, W., Delputte, P.L., Van Gorp,
H., Misinzo, G., Vanderheijden, N., Duan, X., Project website
Nauwynck, H.J. (2010), Porcine reproduct http://www.porrscon.ugent.be/
ive and respiratory syndrome virus entry
into the porcine macrophage, Journal of Keywords
General Virology, 91: 16591667, Review. PRRSV, RNA virus, vaccine, adaptable
inactivated vaccine, vector vaccine, attenu-
Cruz, J.L., Zuiga, S., Becares, M., Sola, ated vaccine, marker vaccine, DIVA sero-
I., Ceriani, J.E., Juanola, S., Plana, J. and logical assay, virulence, pig, antibiot-
Enjuanes, L. (2010), Vectored vaccines ics, pathogenesis, virology, immunology,
to protect against PRRSV, Virus Research, immunobiology.
154:150160.
Coordinator (UGent)
Darwich, L., Gimeno, M., Sibila, M., Diaz, Prof. Dr Hans Nauwynck
I., de la Torre, E., Dotti, S., Kuzemtseva, Laboratory of Virology
L., Martin, M., Pujols, J., Mateu, E. (2011), Faculty of Veterinary Medicine
Genetic and immunobiological diversities Ghent University
of porcine reproductive and respiratory Salisburylaan 133
syndrome genotype I strains, Veterinary 9820 Merelbeke
Microbiology, 150(12):4962. BELGIUM
Hans.Nauwynck@UGent.be
Tian, D., Zheng, H., Zhang, R., Zhuang, J.,
Yuan, S. (2011), Chimeric porcine repro-
ductive and respiratory syndrome viruses
reveal full function of genotype 1 envelope
proteins in the backbone of genotype 2,
Virology, 412:18.
Partners
Department of Molecular and Prof. Dr Luis Enjuanes L.Enjuanes@cnb.csic.es
Cell Biology
Centro Nacional De
Biotecnologa
CSIC
Campus Universidad
Autnoma de Madrid
Darwin 3
Cantoblanco
28049 Madrid
SPAIN

Centre de Recerca en Sanitat Prof. Dr Enrique Mateu enric.mateu@uab.es
Animal (Cresa)
Esfera UAB
Universitat Autnoma De
Barcelona
Bellaterra
08193 Barcelona
SPAIN
Ingenasa Inmunologa y Dr Maria Jose Rodriguez mjrodriguez@ingenasa.es
Genetica Aplicada, SA
C/Hnos. Garca Noblejas, 39.
8
28037 Madrid
SPAIN
Parco Tecnologico Padano Dr Sara Botti sara.botti@tecnoparco.org
CERSA
Via A. Einstein Localita
Cascina Codazza
26900 Lodi
ITALY
Institute of Virology and Dr Nicolas Ruggli nicolas.ruggli@ivi.admin.ch
Immunoprophylaxis (IVI)
3147 Mittelhusern
SWITZERLAND
National Veterinary Institute Prof. Dr Lars Erik Larsen lael@vet.dtu.dk
The Technical University of
Denmark
Blowsvej 27
1790 Copenhagen V
DENMARK
National Veterinary Research Prof. Dr Tomasz Stadejek stadejek@piwet.pulawy.pl
Institute
Department of Swine
Diseases
OIE Reference Laboratory for
PRRS
Partyzantw 57
24100 Pulawy
POLAND
The Roslin Institute and Royal Dr Tahar Ait Ali tahar.aitali@roslin.ed.ac.uk
(Dick) School of Veterinary
Studies
University of Edinburgh
Roslin BioCentre
The Roslin Institute Roslin
Biocentre
Roslin
EH25 9PS
UNITED KINGDOM

Virology Department Dr Trevor Drew Trevor.Drew@ahvla.gsi.gov.uk
Veterinary Laboratories
Agency (VLA)
Woodham Lane
New Haw
Addlestone
KT15 3NB
UNITED KINGDOM
Central Veterinary Institute Dr Annemarie Rebe Annemarie.Rebel@wur.nl
part of Wageningen UR
Edelhertweg 15
8219 PH Lelystad
NETHERLANDS
Department of Swine Infec- Prof. Dr Shishan Yuan shishanyuan@shvri.ac.cn
tious Diseases
Shanghai Veterinary Research
Institute
Chinese Academy of Agricul-
tural Sciences
No 518 Ziyue Road
Minhang District
200241 Shanghai
CHINA
Department of Animal Health, Dr Tung Nguyen tungncvd@gmail.com
Hanoi, Vietnam
Giaiphong No 15
Phuongmai
Dongda
Hanoi 78 844
VIETNAM
Biological R & D Dr Konrad Stadler konrad.stadler@boehringer-ingel-
Boehringer Ingelheim Vet- heim.com
medica GmbH
Binger Strae 173
55216 Ingelheim am Rhein
GERMANY
147
CHAPTER 4.
Influenza
Introduction
Influenza infections of animals have spread of H5N1, which, by spring 2006, had
recently become a great concern for animal affected eastern and central Europe and
and public health. These include the aware- Africa, causing enormous direct and indirect
ness that a swine-origin influenza virus has losses to the poultry industry, wildlife and
become the most recent human pandemic a significant number of human infections,
virus and that avian viruses of different sub- approximately 50 % of which were lethal.
types have caused enormous damage to the
poultry industry and in some cases human The entire scientific community was largely
health issues. unprepared to manage this outburst of
infections, and did not have adequate
Since the late 1990s, outbreaks of avian answers to the questions raised by deci-
influenza caused by low pathogenicity (LP) sion-makers and the industry relating to
H5 and H7 viruses and also by highly patho animal health and public health concerns
genic H5 and H7 viruses have affected the in an evolving eco-epidemiological situ
European poultry population, in some cases ation. Information was lacking on appropri-
causing devastating consequences to the ate control strategies since, at the time, the
industry. In particular, Italy was affected only products which were available were
by HP H5N2, LP H7N1 and H7N3 and by inactivated oil-emulsion vaccines devel-
a severe HP H7N1 which caused between oped for outbreaks in developing countries.
1999 and 2000 death or culling of over In addition, there was the urge to develop
16millionbirds. In 2003, the Netherlands vaccines with a companion diagnostic
was affected by the most devastating H7 test, which enabled the DIVA (differentia-
HP outbreak recorded to date which caused tion of infected from vaccinated animals)
the death or culling of 30millionbirds, strategy, essential to map infection within
approximately 50 % of the entire national avaccinated population. EU-funded efforts
poultry population. This outbreak also such as FLUAID, AIV VACC DIAGNOSIS and
caused several cases of conjunctivitis in Novaduck addressed specifically these
humans and was lethal for one veterinarian objectives, also exploring the efficacy of
involved in the emergency operations to put vaccination in diverse avian species.
the outbreak under control. It was a prel-
ude to the more significant and worrisome Direct control measures are always a crucial
implications of the H5N1 panzootic. component of control and eradication pro-
cedures, as the resistance of viruses in the
The first detection of the latter was traced environment is a key factor that influences,
to a goose in Guangdong province in China in for example, the duration of the cleaning
1996. Infection spread to Hong Kong in 1997, period before restocking. Rivers and FLU-
causing the death or culling of one and a half RESIST have generated significant data on
million birds. Infection appeared to have been environment-related aspects of prevention
eradicated until 2003 when several south- and control, and have highlighted the great
east Asian countries virtually simultaneously variability that exists between viruses, even
notified infection. As infection continued to of the same subtype.
spread in Asia, it also spread north, affect-
ing a mixed wild-domestic bird population at The behaviour of avian influenza within
the Quinghai Lake in late 2005. This was the apoultry flock, the transmission dynamics
dawn of the westward and transcontinental and the effects of vaccination on the spread
C H A P T E R 4 . I N F L U E N Z A 149
of infection were also largely unknown, and FLUTRAIN effort was focused on bridging
this knowledge gap impaired educated deci- gaps across EU borders, with third countries
sion-making in the face of an outbreak, par- mainly in Africa and Central/Eastern Asia.
ticularly in areas in which poultry was raised Specific training was provided to partners
at high densities with multiple species and concerning general information, specific
production categories. The Healthy Poultry areas of interest and transfer of technology.
consortium addressed these issues, analys-
ing retrospectively data that was generated Over the years, the EU has funded two
during the Dutch and Italian outbreaks and surveillance efforts for influenza in pigs
laying down guidelines for future manage- ESNIP2 and ESNIP3. Although swine influ-
ment approaches. enza was not perceived as a major issue
for animal and public health, a forward-
The H5N1 crises highlighted that HPAI avian looking approach allowed the EU to be
influenza viruses can infect a variety of hosts able to collect surveillance data on swine
that were previously believed to be resistant, influenza, which revealed itself as funda-
such as wild birds and some mammals. This mental when the swine-origin H1N1 (2009)
awareness triggered international efforts pandemic emerged. The FLUPIG project
on wild bird surveillance performed by the instead was developed in order to address
Directorate-General for Health and Consum- some scientific questions on the adaptation
ers and others supported by the Directorate- of influenza viruses to the pig and on the
General for Research and Innovation. Among emergence of a pandemic virus for humans.
these New-Flubird, an extensive and compre-
hensive effort to monitor influenza viruses The significant effort made by the Directo-
in wild bird populations across Europe and rate-General for Research and Innovation,
experimental studies in wild birds, has shed guided by Isabel Minguez-Tudelas tireless,
light on previously unexplored areas. The open and flexible approach has placed EU
FLUPATH project was, from certain perspec- scientists in a position of international lead-
tives, a cross-cutting project which aimed at ership. We now have a wealth of knowledge
understanding host-pathogen interactions which, in many cases, has been used abroad
in a variety of avian and non-avian species, and by the industry, and is considered as
including an immunological component to complementary to knowledge generated
answer open issues on the hosts natural and in the United States and by other coun-
induced immune response. tries. Above all, the Directorate-General for
Research and Innovations vision has ena-
The spread of H5N1 to three continents in bled Europe to consolidate existing networks
the western hemisphere, and its circulation and expand them to developed and under-
in under-resourced countries, prompted the resourced countries and thus consolidate its
EU to bridge gaps of knowledge and composition of excellence in scientific research.
munication with countries that were experi
encing massive outbreaks, and required Ilaria Capua
international support. At an EU level, the Director, Istituto Zooprofilattico
FLU-LAB-NET effort acted as a meansby Sperimentale delle Venezie
which EU expertise was made available OIE/FAO Reference Laboratory
within the EU-27 and to neighbouring for Newcastle Disease and Avian Influenza
countries, together with ConFluTech. The Padova, Italy
4.1. Avian flu

[AIV-VACC]
Vaccine, diagnostic test

development and immunology
aspects of avian influenza
Acronym:
Summary birds by serological tests, including an agar
AIV-VACC Avian influenza (AI) is a zoonotic disease gel immunodiffusion test, which is based
and seen as one of the most important on the detection of antibodies against the
Project number:
44141 emerging diseases with serious economic nucleoprotein. However, the recombinant
consequences. Although very useful in the fowlpox-H5 vaccine also requires admin-
EC contribution:
fight against AI, all currently available influ- istration by parenteral injection. Another
EUR 1 372 890
enza vaccines have considerable shortcom- major disadvantage is the fact that in cases
Duration: ings; several vaccines developed over the of previous fowlpox vaccinations, take-up of
48 months
past two decades to protect poultry against the vaccine will be prevented and no immu-
Start date: the highly pathogenic H5 or H7 are based nity against AI will develop. This is another
1 December 2006 on inactivated whole-virus vaccines. Apart serious drawback since fowlpox vaccination
Instrument: from the challenge of setting up a robust is still widely used.
Specific targeted diagnostic test for differentiating infected
research or from vaccinated animals (DIVA principle),
innovation project
(STREP)
these vaccines have to be administered by Problem
labour-intensive and expensive parenteral Inactivated poultry vaccines have disadvan-
injections. In view of the worrying spread of tages related to the application methods
epidemic AI H5N1 in the world and the large and the doses of antigen that needed to
undertaking to vaccinate billions of poultry be used in the vaccine. For mass applica-
in some parts of the world, development of tion in case of an outbreak, these aspects
efficacious vaccines that could be adminis- represent considerable disadvantages that
tered by mass application routes, such as could be overcome with live vector vaccines
spray or drinking water, is greatly needed. which could be administered via more prac-
In the endeavour to develop an improved tical routes such as drinking water, sprays
vaccine against AI, recombinant DNA tech- or eye drops.
nology has been employed to generate vec-
tored, subunit or DNA vaccines. Although
a wide range of these vaccines have been Aim
experimentally shown to be effective vac- The primary aim of this project has been
cines against AI, only a fowlpox-vectored to develop better AI vaccines through live
vaccine with H5 gene insert was commer- or vector vaccines that can be mass appli-
cially available at the start of this project. cable through sprays, drinking water or eye
This recombinant vaccine enables differ- drops. These vector vaccines would offer
entiation between infected and vaccinated considerable advantages:
being mass applicable; virus (NDV) vectors carrying inserts of recent

with less labour-intensive and more isolates of H5N1 influenza virus. Several
animal friendly application; recombinants with an H5 insert were gener-
providing protection by local and sys- ated at different gene junctions and tested in
temic immunity and less interference vitro. All expressed H5 well and no clear dif-
with eventual maternal antibodies; ference of viral replication was observed. But
providing more complete protection these recombinants experienced some inter-
through cellular and humoral immunity; ference from active or passive NDV immu-
faster onset of immunity when used in nity and therefore did not qualify as avac-
the face of an outbreak; cine virus. Another recombinant with better
cheaper production methods. antigenic features was also created with the
potential of inducing a sufficient immune
The project exploited recently acquired response in maternal antibody-positive ani-
knowledge concerning the molecular char- mals. Finally, additional recombinants were
acterisation of the viruses resulting in the generated with the aim to obtain an NDV
construction of candidate strains with backbone with higher virulence. However,
highly interesting efficacy and safety pro- these new NDV/AIV vaccine viruses have yet
file. Safety and efficacy with Newcastle dis- to be tested in animal experiments.
ease (NDV) vectors and Infectious Laryn
gotracheitis (ILT) vectors both for H5 and The objective of WP2 was the characteri-
for H7 inserts have already been demon- sation in vivo of the vector viruses gener-
strated in vivo. A system in which gene cas- ated by WP1 and the selection of the best
settes for the foreign proteins can easily be suitable potential vaccine strains based on
constructed and exchanged would be devel- preliminary safety and efficacy testing. The
oped to be able to respond very quickly to a insertion of the haemagglutinin gene of
change in antigenicity of the field virus. Fur- a highly pathogenic avian influenza virus
ther optimised additional candidate strains H5N1 and its expression by NDV did not
were constructed and extensively tested. increase the virulence of recombinant NDV.
Experiments on genetic in vitro and in vivo The intracerebral pathogenicity index (ICPI)
stability, immunological responses, viru- was 0 compared to 0.3 of the parental vac-
lence testing, spreading, and transmission cine strain. However, a slight enhancement
studies in chickens, ducks and other avian of virulence would be desirable to overcome
species were planned. Such vaccines would maternal-derived antibodies against NDV of
also have marker aspects which will allow MDA+ chickens.
differentiation of infected from vaccinated
animals (DIVA principle). Sensitive, specific The objective of WP3 was to test the effi-
and easy-to-use marker diagnostic tests cacy of NDV-H5 in one-day-old chickens
compatible with the vaccines would also be with different active and passive NDV pro-
developed. tection levels. Previous active immunisation
with NDV does interfere with the take-up
of NDVH5 in chickens when given by spray
Results vaccination. Besides, the efficacy of NDVH5
The activities of the AIV VACC DIAGNOSIS is also negatively influenced by the pres-
project have been divided into eight work ence of maternal-derived antibodies against
packages (WPs). The project, originally NDV. This interference could be related to the
planned for three years was extended to low virulence of the NDVH5 vector virus. So,
afourth and final year. efforts were undertaken to enhance the viru-
lence of the vector backbone. It was decided
The objective of the first WP was the con- to broaden the vector scope, and to include
struction of optimised Newcastle disease work on Infectious Laryngotracheitis (ILT)
virus, another avian influenza vector virus In Europe, regulatory approval was obtained
with proven vector capacities and AI efficacy in Germany. The test was also validated by
data. But, later on, ILTV vaccines were aban- GD Deventer (Netherlands).
doned from the market. So other NDVH5 vec-
tor viruses were prepared and evaluated but The objective of WP6 was to develop alter-
none had the desired properties for a vaccine native and optimised tissue culture pro-
strain, being too virulent for chickens. How- duction systems that would create inde-
ever, in the meantime, a new more promising pendency of AI vaccine production from
construct (rNDVGu) was developed in WP1 egg-based production. A variety of cell
that should have better properties in chick- lines was studied and although some cell
ens. The strain NDV Gustav induces a solid lines could be used, none tested gave titres
protection against challenge with the NDV comparable to the ones obtained in eggs.
Beaudette virus, only slightly different from Due to the uncertainty regarding the choice
that induced by the wild type NDV. So, vector of the actual vector, work continued using
viruses using NDV Gustav as the backbone the parental NDV Clone30, a vaccine strain
and inserting the genes encoding for the itself. A proprietary suspension-cell plat-
high or low pathogenic H5 of the Vietnamese form was chosen as the preferred cell sub-
H5N1 isolate, were constructed. strate for production but found to be (much)
less potent than egg-based material.
The objective of WP4 was the performance
of necessary testing for vaccine devel- The objective of WP7 was to gain an
opment and environmental risk assess- improved understanding of the immunologi-
ment purposes in avian and other species. cal pathways after vaccination with AI vac-
ANDVH5 master virus seed, working virus cines or infection with AI in chickens: immu-
seed and production virus seed were pro- nophenotyping of vector-induced immunity
duced and NDVH5 is genetically stable on against AI. Primers and the correspond-
passages through embryonated eggs. This ing probe sets were designed and partially
was confirmed by sequencing the sequences tested allowing the evaluation of Th-1 and
flanking the H5 gene and the flanking F and Th-2 immune responses as well as CTL
HN genes. However, no further work has activity. The quantitative PCR procedures
been conducted according to the original WP were tested on in-house lung tissue materi-
description due to the poor efficacy of avail- als, isolated from earlier animal experiments
able virus vectors in MDA+ chickens and in involving a respiratory influenza (H9N2)
chickens actively immunised against NDV. challenge. A new primer and corresponding
probe (chicken (ch) CD40) were designed and
The objective of WP5 was to develop and tested as possible B cell markers. In addi-
validate diagnostic marker tests that can be tion two more sets were developed: ChTGF-
used in conjunction with the marker vaccines (Th3/Treg marker) and Ch28S (a new refer-
or conventional vaccines. A large collection ence marker). Finally, the Taqman assay was
of negative and positive serum samples started for ChIFN- and ChIFN-.
from different avian species and from dif-
ferent geographical regions has been estab- The objective of WP8 was to develop meth-
lished. Test reagents (proteins and monoclo- ods allowing a quick response to changes in
nal antibodies) were developed and used for antigenicity of the field-virus, and construc-
assay optimisation. A test, the FlockChek tion of a set of vectors containing other hae-
Avian Influenza MultiS-Screen Antibody Test, magglutinin subtypes (e.g. H7, H9) for vacci-
was developed, validated and approved by nation. A NDVH5 vector backbone in which an
USDA for five species: chicken, turkey, duck, H5 can easily be substituted by another hae-
goose and ostrich. The kit is now available on magglutinin was constructed: transcription
the market in the United States and Europe. cassettes were inserted at three different
NDV gene junctions. The difference of the H5 constructs have become available. Thus,
expression level was unexpectedly low, the promising avenues for further experiments
insertion site is not crucial for the expression have been identified and will be followed
of the foreign gene. However, because of the after this FP6 project has ended, including
improved characteristics of the new recombi- work on improved cell culture systems for
nant NDV (rNDVGu), further constructs shall the production of sufficient virus titres.
be generated and experiments performed
when rNDVGu excites a sufficient antibody As stated above, avian influenza is seen as
response in MDA+ chickens. one of the most important emerging zoonotic
diseases with potentially serious economic
consequences, both in the industrialised
Potential applications world and in developing countries. Poultry
The final aim of this project was to develop farming, whether it be in an industrial set-
a H5N1 vaccine that is mass applicable ting, for example, in Europe or the small-
though sprays, drinking water or eye drops scale backyard poultry farming customary
on the basis of recombinant NDVH5 or in developing countries, as well as wildlife,
other recombinant vector viruses. In add must be considered a potential source of a
ition, to be able to respond very quickly to serious worldwide influenza outbreak. There-
a change in antigenicity of the field virus, fore, there is an absolute need for more effi-
asystem in which gene cassettes for the cacious measures to combat this virus, for
foreign proteins can easily be constructed instance measures via more efficient vacci-
and exchanged, was to be developed. The nation strategies. A well-vaccinated popula-
development of a system that allows propa tion provides a proven barricader, preventing
gation of virus vaccine in cell lines instead transmission of the virus. To date, an efficient
of eggs was started, in order to contribute and effective, mass applicable, AI vaccine
to decreasing production costs. Finally, an that could be used in either of the settings
ELISA-based assay would be developed mentioned above is lacking. In addition, there
that would enable discrimination between is demand for vaccines that allow a clear dif-
infected and immunised avian species. ferentiation between infected and vaccinated
animals (DIVA principle).
Although the mass applicable H5N1 vac-
cine has not been developed yet, consider- The AIV VACC DIAGNOSIS project has
able progress has been made with respect brought together a unique European part-
to the development of an effective vaccine nership with expertise in the development
vector backbone based on the wild-type of diagnostics (Bommeli-IDEXX), AI vac-
NDV Clone30 sequence (i.e. rNDVGu). With cine research and development (Intervet)
its improved properties concerning replica- and molecular biology of AI and NDV (FLI).
tion and virulence, this vector will be the Together, these partners have achieved
focus of experiments continuing after this considerable progress towards the aim of
FP6 project has ended. developing an efficient and effective, mass-
applicable AI vaccine. In addition, the tools
The expression cassettes that have been that have been developed thus far will allow
constructed during the course of the project, the DIVA principle to be put into practice,
as well as the ELISA-based assay, which has and include an ELISA-based assay that has
meanwhile been commercialised, will be used meanwhile been commercialised.
as tools in further investigation and optimi-
sation of its properties. Safety and efficacy The project has therefore provided a strong
testing, which has been delayed due to the basis for continued research and develop-
lack of proper vaccine vector candidates for ment in the field of avian influenza vaccine
testing, will be possible once the appropriate development, both in an academic and an
industrial setting. However, not all the goals Brown, J.D., Stallknecht, D.E., Berghaus,
of this FP6 project have been achieved: R.D., Luttrell, M.P., Velek, K., Kistler, W.,
therefore, as is indicated above, work on Costa, T., Yabsley, M.J., Swayne, D., Evalu-
reaching these goals is ongoing. ation of a Commercial Blocking Enzyme-
Linked Immunosorbent Assay To Detect
Avian Influenza Virus Antibodies in Mul-
References/publications tiple Experimentally Infected Avian Spe-
Veits, J., Rmer-Oberdrfer, A., Helfer- cies, Clinical and Vaccine Immunology,
ich, D., Durban, M., Suezer, Y., Sutter, G., (2009)16, 824829.
Mettenleiter, T.C., Protective efficacy of
several vaccines against highly pathogenic Ramp, K., Skiba, M., Karger, A., Metten-
H5N1 avian influenza virus under experi- leiter, T.C., Rmer-Oberdrfer, A., Influ-
mental conditions, Vaccine, (2008) 26, ence of Insertion Site of Avian Influenza
16881696. Virus Hemagglutinin (HA) Gene Within
the Newcastle Disease Virus Genome on
Rmer-Oberdrfer, A., Veits, J., Helferich, HA Expression, Journal of General Virol-
D., Mettenleiter, T.C., Level of protection ogy, first published on 10 November 2010
of chickens against highly pathogenic (doi:10.1099/vir.0.027268-0 92: 361369).
H5 avian influenza virus with Newcastle
disease virus based live attenuated vec- Keywords
tor vaccine depends on homology of H5 avian influenza, viral pathogens of animals,
sequence between vaccine and challenge animal immunology, animal diagnostics
virus, Vaccine, (2008) 26, 23072313.
Coordinator
Schrer, D., Veits, J., Grund, C., Dauber, Intervet International B.V.
M., Keil, G., Granzow, H., Mettenleiter, T.C., Wim de Korverstraat 35,
Rmer-Oberdrfer, A., Vaccination with 5830 AA Boxmeer,
Newcastle Disease Virus Vectored Vaccine NETHERLANDS
Protects Chicken Against HighlyPathogenic (Contract Manager/
H7 Avian Influenza Virus, Avian Diseases, Coordinator Dr Danny Goovaerts)
53 (2009) accepted. Danny.Goovaerts@sp.intervet.com
Partners
Friedrich-Loeffler-Institut, Friedrich-Loeffler-Institut Thomas.Mettenleiter@fli.bund.de
Federal Research Institute for (contract manager Prof.
Animal Health, Dr Thomas Mettenleiter)
Boddenblick 5a,
17493 Greifswald-Insel Riems,
GERMANY
Bommeli Diagnostics, Sta- Bommeli Diagnostics Christian-Schelp@idexx.com
tionsstrasse 12, CH-3097 (contract manager Dr
Liebefeld, SWITZERLAND Christian Schelp)
[FLUAID]
Generation of information
and tools to support the
management of the avian
influenza crisis in poultry
Summary millions of dead animals and raising con-
Acronym:
Avian influenza (AI) has become a great cerns regarding the loss of human lives and FLUAID
risk both for animal and human health. By the future management of the pandemic
Project number:
bringing together both European and non- potential of this virus.
022417
European laboratories, the FLUAID pro-
ject aimed to generate data on significant EC contribution:
EUR 1 200 000
issues relating to AI outbreak management Aim
where scientific knowledge at the time of FLUAIDs primary goals were: Duration:
the proposal submission was seen to be 46 months
lacking. to improve scientific knowledge on AI; Start date:
the joint development and applica- 1 January 2006
tion of novel technologies to combat AI Instrument:
Problem infections. Specific Targeted
Between 2000 and 2005, avian influenza Research or
outbreaks caused severe losses to the poul- The goals were achieved through the inter- Innovation Project
(STREP)
try industry, its stakeholders and, ultimately action of leading European institutes along
to the EU taxpayer. It is estimated that 200 with the active collaboration of laboratories
million birds died or were culled following in Egypt, Indonesia, Pakistan, South Africa
infection with influenza viruses subtypes and Vietnam.
H5 or H7. Approximately 50 million of these
birds were from Europe. Most importantly,
human infections were also reported in Results
several of these outbreaks. On a global In addition to identifying possible strains for
level, H5N1 outbreaks continue to be a seri- an EU vaccine bank and producing new com-
ous concern for food security and human panion diagnostic kits, FLUAID researchers
health with the crossing of the species bar- also improved knowledge on pathogenesis
rier representing a serious potential risk of and the transmission of the AI virus in indi-
a new human pandemic virus emerging. The vidual bird species (e.g. native chickens, tur-
increased relevance of AI in the fields of keys, mule and Muscovy ducks). Of particular
animal and human health has highlighted note was the age-related association with
the lack of scientific information available infection in Pekin ducks, with the data gen-
on several aspects of the disease. This erated suggesting that quail may act as a
has hampered the adequate management silent reservoir of AI infection. In generating
of some of the recent crises resulting in this data within FLUAID, the consortium
promoted European expertise, know-how the pathogenesis of a highly pathogenic

and scientific achievement globally. avian influenza (HPAI) H5N1 virus in Pekin
(Anas platyrhynchos) ducks infected experi-
mentally, Influenza and Other Respiratory
Potential applications Viruses, 4, 1725.
Using monoclonal antibodies generated
within the consortium, three companion Salzberg, S.L., Kingsford, C., Cattoli, G.,
diagnostic kits were produced with one Spiro, D.J., Janies, D.A., Aly, M.M., Brown,
of these commercialised successfully. I.H., Couacy-Hymann, E., De Mia, G.M., Dung,
Using phylogenetic analysis and sero- D.H., Guercio, A., Joannis, A., Maken Ali, A.S.,
logical data, suitable viral candidates for Osmani, A., Padalino, I., Saad, M.D., Savi,
a European vaccine bank were identified. V., Sengamalay, N.A., Yingst, S., Zaborsky,
Reassortant viruses with rare neuramin J., Zorman-Rojs, O., Ghedin, E., Capua, I.
idases that could be used as vaccines as (2007), Genome analysis links recent Euro-
part of a DIVA (differentiation of infected pean and African H5N1 influenza viruses,
from vaccinated animals) strategy were Emerging Infectious Diseases, 13: 7138.
generated.
Protocols for the identification of the H7 Toffan, A., Beato, M.S., De Nardi, R., Bertoli,
and H5 subtype viruses were success- E., Salviato, A., Cattoli, G., Terregino, C.,
fully transferred to mobile molecular Capua, I. (2008), Conventional inacti-
diagnostic platforms. vated bivalent H5/H7 vaccine prevents viral
localisation in muscles of turkeys infected
experimentally with LPAI and HPAI H7N1
isolates, Avian Pathology, August 2008,
37(4): 40712.
van der Goot, J.A., van Boven, M., Stegeman,

A., van de Water, S.G., de Jong, M.C., Koch, G.
(2008), Transmission of highly pathogenic
avian influenza H5N1 virus in Pekin ducks is
significantly reduced by a genetically distant
H5N2 vaccine, Virology, 382(1): 917.
Keywords
avian influenza, vaccines, diagnostics
Coordinator
Ilaria Capua DVM PhD
OIE/FAO and National Reference Laboratory
References/publications for Avian Influenza and Newcastle Disease
Bos, M.E., Nielen, M., Koch, G., Stegeman, A., Istituto Zooprofilattico Sperimentale delle
De Jong, M.C. (2008), Effect of H7N1 vac Venezie
cination on highly pathogenic avian influ- Legnaro (PD)
enza H7N7 virus transmission in turkeys, ITALY
Vaccine, 25, 26: 63228. icapua@izsvenezie.it
Lndt, B.Z., Nunez, A., Banks, J., Alexan-

der, D.J., Russell, C., Richard-Lndt, A.C.,
Brown, I.H. (2009), The effect of age on
Partners
Veterinary Laboratories Agency Dr Jill Banks j.banks@vla.defra.gsi.gov.uk
Virology Dept,
Woodham Lane
Addlestone, KT15 3NB, Surrey
UNITED KINGDOM
Central Veterinary Institue (CVI) Dr Guus Koch Guus.Koch@wur.nl
Houtribweg 39,
8221 RA Lelystad
NETHERLANDS
Agence Franaise de Scurit Sanitaire Dr Vronique v.jestin@ploufragan.afssa.fr
des Aliments, AFSSA Jestin
site de Ploufragan, B.P. 53
22440 Ploufragan
FRANCE
National Agricultural Research Centre Dr Khalid Naeem naeem22@isb.comsats.net.pk
Park Road, Islamabad
PAKISTAN
Institute of Virology and Immuno- Dr Kenneth Kenneth.McCullough@ivi.admin.ch
prophylaxis Sensemattstrasse McCullough
293 Mittelhaeusern, SWITZERLAND
CSIRO Livestock Industries, Australian Dr Paul Selleck Paul.Selleck@csiro.au
Animal Health Laboratory
P.O. BAG 245, Portarlington Road,
Geelong
AUSTRALIA
Forsite Diagnostics Ltd. Mr Chris Danks c.danks@ForsiteDiagnostics.com
Sand Hutton, York YO41 1LZ
UNITED KINGDOM
Innovative Diagnostic, Vet 1682 Mr Philippe philippe.pourquier@id-vet.com
Rue de la Valsire Pourquier
340810 Montpellier
FRANCE
Dept of Biochemistry Prof. Dirk Uwe dub@sun.ac.za
University of Stellenbosch, Bellstedt
Box X1
1 Victoria St, 7602 Stellenbosch
SOUTH AFRICA
Directorate of Animal Health, Disease Dr Isep Sulaiman
Investigation Centre
JL Raya Yogya Wates Km 27,
Wates Yogyakarta
INDONESIA
National Center for Veterinary Dr Thanh Long To thanhto@fpt.vn
Diagnosis,
Department of Animal Health
1178th Lane, Giai-Phong,
St Dong-da, Hanoi
VIETNAM

Central Laboratory for Veterinary Dr Mona Aly monaaly5@yahoo.com
Quality Control on Poultry Production,
Nadi El-Seid Street, Dokki, Giza,
PO Box 264-Dokki
EGYPT
[Novaduck]
Novel AI DIVA recombinant

vaccines for duck
Acronym: Summary Problem
Novaduck The Novaduck project gathered eight pub- The ongoing outbreak of H5N1 HPAI has
lic and private partners that aimed to spread from Asia to Europe and Africa and
Project number:
044217 develop and evaluate new avian influenza become endemic in several countries, pos-
live vaccines for ducks based on live vec- ing a real public threat as HPAI can occa-
EC contribution:
tors and compatible with the DIVA strat- sionally infect humans. Ducks play a major
EUR 1 416 380
egy (differentiation of infected from vac- role in the epidemiology of avian influenza
Duration: cinated animals). Viral vectors have been because wild waterfowl, including ducks,
39 months
engineered to express the best protective constitute the natural reservoir of all sub-
Start date: gene selected in a DNA vaccination model. types of influenza A virus. Experimental
1 January 2007 Two types of vectors (fowlpox and Newcas- infection of ducks with recent isolates indi-
Instrument: tle disease virus (NDV)) have been evalu- cates a longer shedding period and a selec-
Specific Targeted ated for safety and immunogenicity using tion for lower virulence variants, suggesting
Research or newly developed duck-specific immunologi- that duck has become the Trojan horse of
Innovation Project
(STREP)
cal tools. Their efficacy was also assessed Asian H5N1 AI.
against highly pathogenic avian influenza
(HPAI) H5N1 challenge in both Pekin and Although biosecurity is the first line of
Muscovy ducks. The Novaduck project has defence against HPAI, strategic use of vac-
demonstrated, for the first time, the proof- cination is clearly recognised as a tool to
of-concept of the use of a NDV-based help eradicate HPAI in an infected country.
vector vaccine administered once by a Most studies evaluating the efficacy of AI
non-parenteral route in day-old Pekin or vaccines have been performed in chickens,
Muscovy ducklings. The level of protection and duck studies have been relatively rare.
against HPAI H5N1 was further improved by Existing inactivated AI vaccines are less
a prime-boost regimen involving two differ- immunogenic in ducks than in chickens and
ent viral vectors. The compatibility of these must generally be administered twice to
vaccines with the DIVA strategy has been be fully efficient; furthermore, there is no
confirmed and the effect of vaccination commercially available DIVA test to monitor
on genetic and antigenic drift of H5N1 has AI infection in birds injected with this type
been assessed. The Novaduck project suc- of vaccine. Therefore, highly efficient, cost-
ceeded in achieving most of its initial objec- effective, DIVA-compatible AI vaccines for
tives and valorisation of results is ongoing. ducks are still greatly needed.
In this specific context, live vector-based candidates against recent highly patho-
vaccines hold the greatest promise and are genic avian influenza (HPAI) H5N1 and
one of the most effective options. Indeed, compare it with existing vaccines;
some live recombinant vector-based AI vac- to study the effect of vaccination on
cines have shown excellent results in chick- genetic/antigenic drift;
ens, but they are not necessarily adapted to select the best candidate(s) to be
for use in ducks. Expected advantages of developed based on its (their) immu-
this type of vaccine include administration nogenic and protective properties as
at a younger age, mass administration, well as its (their) estimated cost of
rapid onset of immunity, and compatibility production and administration mode
with the DIVA strategy. flexibility (e.g. individual versus mass
administration).
The Novaduck project was designed to dem-
onstrate and exploit the potential of live Results
vector vaccines to develop a new generation An optimal protective gene was selected
of highly efficient and cost-effective AI vac- using a needle-free DNA vaccination
cines for ducks and therefore could contrib- model in SPF Muscovy ducks. The immune
ute to decreasing AI from the ecosystem. response induced by plasmid DNA able to
generate retrovirus-based virus-like par
ticles (VLPs) was also evaluated in both
Aim mice and Muscovy ducks. This optimal gene
The Novaduck project aimed to develop and was inserted in different viral vectors includ-
evaluate new, highly protective and cost- ing poxvirus. The safety of three different
effective avian influenza live vaccines for poxvirus vectors expressing the same HPAI
ducks based on live vectors and in line with H5N1 HA (haemagglutinin) gene was found
the DIVA strategy. to be excellent in Muscovy ducks. However,
the immune humoral response induced by
More specifically, the main objectives of the these pox vectors in ducks appeared to be
Novaduck project were: low and transient. A significant increase of
immunogenicity was obtained when the pox
to identify the optimal avian influenza vector-induced primary immune response
(AI) immunogenic sequence(s) to be was boosted by an inactivated vaccine
inserted into the selected live vectors; and the intensity of the boost effect was
to generate and optimise three types of dependent on the dose of the pox vector.
live recombinant vector-based vaccines;
to develop reliable and cost-effective The optimal protective gene was also
duck-specific immunological tools to expressed in an avian paramyxovirus 1
measure the immune response induced (APMV1, better known under the name
by the different vaccine candidates and of Newcastle disease virus (NDV)) vector.
to detect infection in a vaccinated duck The HA was shown to be expressed at the
(DIVA strategy); surface of NDV virions. The safety of this
to assess the safety and immunogen NDV-based vector vaccine was excellent
icity of the new vectored vaccine can- and the AI HA surface expression did not
didates and compare these with those alter the tissue tropism of the NDV vec-
of existing vaccines to select the best tor in both Pekin and Muscovy ducks. The
vaccine candidate(s); humoral immune response induced by the
to set up a challenge model in ducks for NDV vector candidate when administered
vaccine evaluation of efficacy; by a non-parenteral route at day-of-age
to measure the efficacy of the most ducklings was very low or undetectable
immunogenic vec tored vaccine by the HI test or an H5 blocking ELISA; the
seroneutralisation test was found to be analysis of H5N1 virus recovered from ducks
slightly more sensitive but antibody titres vaccinated and challenged did not show any
were low. significant antigenic or genetic drift.
Immunological tools were developed to In summary, the Novaduck project suc-

evaluate the duck immune response. Sev- ceeded in achieving most of its initial
eral monoclonal antibodies recognising objectives. The NDV-based vector vaccine
the duck interferon gamma (DuIFN) were was shown to be safe and it induced a high
generated but they were unfortunately level of H5N1 protection in all bird species
unable to detect natural DuIFN produced tested after only one non-parenteral admin-
by mitogen-activated splenocytes. A real istration at day-of-age. A heterologous
time RT-PCR to quantify DuIFN mRNA was prime-boost regimen using two different
then developed and could better detect vectors further improved the level of pro-
duck cell-mediated immune (CMI) response tection. Further improvement of duck-spe-
than the HD11 cell-based bioassay. AIV- cific immunological tools will be needed to
specific IgA, IgM, and IgY ELISAs were also reproducibly detect the protective immune
developed to evaluate the humoral as well response induced by one administration of
as the mucosal immune response. These the NDV-based vector.
duck-specific immunologicals remained
not sensitive enough to reproducibly detect
an immune response induced by one Potential applications
administration of the NDV-based vector. The work performed within Novaduck has
demonstrated for the first time the proof-
HPAI H5N1 challenge models were devel- of-concept of the use of a NDV-based
oped in Muscovy, Pekin and mule ducks vector vaccine in day-old ducklings. This
and the Pekin and mule ducks were found vector allows the mass application of vac-
to be much more resistant than Muscovy cine at the hatchery and is compatible
ducks. The fowlpox/inactivated prime/boost with the DIVA strategy. Additional stud-
vaccination scheme was shown to induce ies will be needed to confirm the efficacy
a good level of protection after H5N1 chal- profile of the selected vector candidates
lenge in both Muscovy and Pekin ducks. against other epidemiologically relevant
However, the use of the inactivated vaccine H5N1 isolates and in presence of mater-
suppressed the possibility to use the DIVA nally-derived anti-vector and/or anti-AI
strategy based on commercially available antibodies before entering full develop-
NP-based ELISA. The NDV-based vector ment. The prime-boost vaccine regimen
vaccine induced a high level and duration of consisting of a prime with fowlpox vector
H5N1 protection in all three types of ducks and a boost with inactivated vaccine can
after only one non-parenteral administration now be evaluated in field conditions since
at day-of-age in spite of non-detectable or both fowlpox and inactivated vaccines are
very low antibody response. To our know commercially available. The immunology
ledge, this is the first time that a vaccine tools developed for ducks can be used to
administered by a non-parenteral route at evaluate the immune response induced by
day-old is shown to induce such level of other vaccines in this species. NP-based
protection in birds. A heterologous prime- ELISAs have been thoroughly evaluated on
boost regimen using two different vectors a panel of duck sera and were shown to be
further improved the level of protection. In valuable as DIVA tests in birds vaccinated
addition, we confirmed that this prime-boost with vector vaccines. HPAI H5N1 challenge
combination of vector vaccines was compat- models have been developed in different
ible with the DIVA strategy using internal or duck types and will be useful tools to eval-
commercial NP-based ELISA. The sequence uate current and new AI vaccines for ducks
and to define the standard for evaluation Infection with H5N1 and H7N1 Low Patho-
of AI vaccine efficacy. genic Avian Influenza Viruses, Avian Dis-
eases, Vol. 54, No 1, Supplement 2010,
Overall, the Novaduck project has greatly pp.660667.
increased the knowledge of the immune
response of ducks, H5N1 challenge models Steensels, M., Bublot, M., Van Borm, S., De
for the different types of ducks and the Vriese, J., Lambrecht, B., Richard-Mazet, A.,
AI vaccine evaluation and performances Chanavat-Bizzini, S., Duboeuf, M., Le Gros,
to be expected in ducks. Some results of F.-X., van den Berg, T., Prime-boost vacci-
the Novaduck project have already been nation with a fowlpox vector and an inac-
published in peer-reviewed scientific jour- tivated avian influenza vaccine is highly
nals and in another scientific bulletin. Ten immunogenic in Pekin ducks challenged
abstracts have been presented at seven with Asian H5N1 HPAl, Vaccine, Vol. 27,
international scientific congresses and Issue 5, 29 January 2009, pp. 646654.
results of the projects have been presented
at 13 local scientific meetings and at sev- Project website
eral seminars with stakeholders. Additional http://www.novaduck.eu
publications containing the latest data on
efficacy are being prepared and an exploita- Keywords
tion plan has been initiated to valorise the avian influenza, recombinant vaccines,
results of the project. vectored vaccines, ducks, DIVA, immune
response
References/publications Coordinator
Bublot, M., Richard-Mazet, A., Chanavat- Dr Michel Bublot
Bizzini, S., Le Gros, F.-X., Duboeuf, M., Stoll, Merial SAS
A., Plfi, V., Guionie, O., Niqueux, E., Dren, N., 254 rue M Mrieux
Immunogenicity of Poxvirus Vector Avian 69007 Lyon
Influenza Vaccines in Muscovy and Pekin FRANCE
Ducks, Avian Diseases, Vol. 54, No 1, Sup- Tel. +33 472725973
plement 2010, pp. 232238. michel.bublot@merial.com
Bublot, M., La vaccination antigrippale chez

lanimal, Bulletin et Mmoires de lAcadmie
royale de Mdecine de Belgique, Vol. 164,
Anne 2009, No 10.
Guionie, O., Guillou-Cloarec, C., Courtois, D.,

Bougeard, S., Amelot, M., Jestin, V., Experi-
mental Infection of Muscovy Ducks with
Highly Pathogenic Avian Influenza Virus
(H5N1) Belonging to Clade 2.2., Avian Dis-
eases, Vol. 54, No 1, Supplement 2010,
pp.538547.
Lage Ferreira, H., Pirlot, J.F., Kaspers, B.,

Kothlow, S., van den Berg, T., Lambrecht, B.,
Development of Specific Enzyme-Linked
Immunosorbent Assays to Evaluate the
Duck Immune Response After Experimental
Partners
Agence National de Scurit Dr Vronique Veronique.JESTIN@anses.fr
Sanitaire (ANSES) Jestin
Laboratoire de Ploufragan-Plouzan
BP 53 Zoople
22440 Ploufragan
FRANCE
Veterinary and Agrochemical Dr Thierry thvan@var.fgov.be
Research Centre (VAR) Vandenberg
Dept of Small Stock
Diseases, Avian Virology and
Immunology Unit
Groeselenbergstraat 99
1180 Brussels
BELGIUM
Veterinary Medical Research Dr Csaba Dren nickdren@gmail.com
Institute of the HAS
Avian Immunology and Tumour
Virology
Hungria krt 21
PO Box 1581 Pf 18
Budapest
1143
HUNGARY
Central Agriculture Office Veterinary Dr Vilmos Plfi palfiv@oai.hu
Diagnostic Directorate
Dept of Virology
Tbornok utca 2.
Budapest
1149
HUNGARY
The Secretary of State For Dr Jill Banks j.banks@vla.defra.gsi.gov.uk
Environment, Food and Rural
Affairs
Virology Dept Veterinary
Laboratories Agency
Woodham Lane, New Haw
Addlestone KT15 3NB
UNITED KINGDOM
Diasource Fabienne Mathieu Fabienne.Mathieu@diasource.be
Rue de lIndustrie 8
1400 Nivelles
BELGIUM
Epixis S A Charlotte Dalba cd@epixis.com
1618 rue de la Glacire
75013 Paris
FRANCE
[Healthy Poultry]
Development of new integrated

strategies for prevention,
control and monitoring of
epizootic poultry diseases
Summary groups: aplatform of experts and deci-
Acronym:
When highly pathogenic strains of influenza sion-makers (the users of the project Healthy Poultry
break out in poultry, the consequences can results), and representatives of major
Project number:
be devastating. Destruction of infected stakeholders in the poultry business who
513737
birds is the primary means of control today would be most affected by the implemen-
but even such drastic measures cannot tation of new policies. Close cooperation EC contribution:
EUR 1 119 404
totally prevent disaster. Given the global between these panels and the project
spread of the H5N1 AI subtype, policymak- research teams could quickly lead to new Duration:
ers across Europe are looking at revising epizootic disease policies that could cir- 45 months
their strategies to prepare for future out- cumvent disaster if H5N1 ever gets into Start date:
breaks of epizootic diseases in poultry: that poultry stocks. 1 November 2004
is how to best avoid infection and, if infec- Instrument:
tions occur, how best to limit their spread Specific Targeted
and impact. Problem Research or
The EU aims at assuring a high level of Innovation Project
(STREP)
Healthy Poultry aims to assess current animal health and animal welfare with-
scientific understanding of epizootic AI. New out compromising the functioning of
strategies will be suggested for their pre- the internal market. Nevertheless, in
vention, control and monitoring. Guidelines the last decade, several epizootics of AI
will be provided for the implementation of occurred throughout the EU. These had
these strategies in EU Member States for a devastating veterinary and economic
specific situations at the regional level. impact. Fear amongst the population
increased because of a possible impact
The partners will complement their recom- on human health, particularly during the
mendations with a risk assessment toolkit. last couple of years. Finally, control of AI
A GIS system will allow policymakers to currently coincides with severe problems
evaluate the possible consequences of dif- related to socio-ethical issues and animal
ferent strategies on the health of poultry welfare.
flocks, the geographical spread of a dis-
ease and economic outcomes of particular Intensive trade contacts (of animals and
interventions. poultry products) between Member States
pose considerable risks to poultry in the
The results of this project will be EU once a single Member State is struck
disseminated through two important by AI.
Strategies and measures for prevention and a GIS-based toolbox for spatial, struc-
control of AI need improvement to fulfil EU tural, demographic and basic disease
objectives. Future prevention and control of risk analysis;
AI should be more efficient, ethically accept- a spatial and geo-statistical analysis of
able and less costly. Self-evidently, because poultry production.
of the single market context of EU livestock
production, only a comprehensive approach Epidemiological aspects:
at the level of the EU is likely to be suc-
cessful. Healthy Poultry aims at addressing an epidemiological analysis of Italian
these issues. data on AI;
an epidemiological analysis of Dutch
data on AI.
Aim
The primary aim of the project is to provide Strategy and economic aspects:
scientifically-based support to decision-
makers in the field of epizootic poultry an analysis of monitoring systems for AI;
disease prevention and control. a qualitative regional risk assessment
for AI;
The objectives of the project are to: an epidemiological-economic analysis of
prevention and control strategies for AI;
develop new integrated strategies for t he pr ov isio n of g uid elin e s for
prevention, control and monitoring of management of AI.
epizootic poultry diseases;
a n a ly s e t h e s e s t r at e g ie s i n a Potential applications
comprehensive way; The primary field of application of the
provide guidelines for the implementa- results is policy and decision-making with
tion of these strategies in EU Member regard to prevention and control of epizo-
States; otic poultry diseases (i.e. AI) particularly at
develop user-friendly toolboxes for the level of the EU and of Member States.
strategy evaluation; Some of the results will also be valuable
disseminate project results to a broad for other stakeholders within the poultry
relevant audience. production chains, for example integrated
production chains, animal health services,
Results product boards.
The main results of the project can be
summarised as follows per task.
References/publications
Spatial and organisational aspects: The full scientific report is available on
the projects website: there, a link can also
spatial parameters and conversion be found to the publications originating
tables; fromthe project and the corresponding
a n orga nisational a nd economic authors.
database;
a farm economic analysis of poultry Project website
production; http://www.healthy-poultry.org/
a spatial, structural and demographic
database of poultry production; Keywords
a database on migratory birds issues; avian influenza, epizootic poultry diseases,
a descriptive analysis of migratory birds disease control and prevention, policy and
issues; decision-making
Coordinator
Dr Helmut Saatkamp
Wageningen University
Dept of Social Sciences
Business Economics Chair
(formerly Farm Management Group)
Hollandseweg 1
6706 KN Wageningen
NETHERLANDS
Tel. +31 317482232
helmut.saatkamp@wur.nl
Partners
University of Vechta, ISPA Hans-Wilhelm hwindhorst@isp.uni-vechta.de
PO Box 1553 Windhorst
49377 Vechta
GERMANY
IZSVe Stefano smarangon@izsvenezie.it
Viale dellUniversit 10, Marangon
35020 Legnaro (PD),
ITALY
Utrecht University, Arjan Stegeman j.a.stegeman@uu.nl
PO Box 80 163,
3508 TD Utrecht,
NETHERLANDS
LEI Peter van Horne peter.vanhorne@wur.nl
PO Box 35,
6700 AA Wageningen,
NETHERLANDS
Szent Istvan University, Oszkar Biro obiro@univet.hu
Faculty of Veterinary Science
Pb. 2
Budapest
1400
HUNGARY
INFS Vittorio Guberti vittorio.guberti@infs.it
Via CaFornacetta 9,
40064 Ozzano E BO
ITALY
[FLUPATH]
Avian influenza: Impact

of virus-host interactions
on pathogenesis and ecology
Acronym: Summary chicken immunology will use multidiscip
FLUPATH Highly pathogenic avian influenza viruses linary and complementary approaches to
(HPAIV) have acquired the unprecedented address key problems and unanswered
Project number:
044220 and alarming capacity to infect humans. questions with respect to the ecology and
By establishing a permanent ecological pathogenesis of avian influenza.
EC contribution:
niche in wild birds, HPAIV will pose a con-
EUR 1 915 800
tinuous risk for poultry and fatal human FLUPATH will provide knowledge and tools
Duration: infections, especially if these birds excrete for new strategies which will be tailored
42 months
HPAIV without showing any clinical signs of for the control and management of avian
Start date: disease. These changes in the ecology of influenza at the European and international
1 January 2007 the disease and behaviour of the virus may level. This will limit the impact of the dis-
Instrument: create opportunities for a pandemic virus to ease both in terms of human health and
Specific Targeted emerge. losses to the poultry industry. The accom-
Research or panying reduction in animal slaughter and
Innovation Project
(STREP)
Attempts to avoid or contain HPAIV out- financial and economic losses will place
breaks have been largely unsuccessful. This a significantly lower demand on EU and
can be directly linked to our lack of funda- Member States budgets.
mental knowledge. Therefore, it is essential
to increase our knowledge of the ecology
and pathology of avian influenza virus Problem
infections in poultry and other species. Avian influenza represents one of the major
concerns for public health that has recently
Full understanding of the ecology and emerged from the animal reservoir. The
pathogenesis of HPAIV requires a multi- increased relevance of avian influenza in
disciplinary approach determining host- the fields of animal and human health has
pathogen interactions and the role played highlighted the lack of scientific informa-
by the host immune response. To this end, tion on the disease. This has hampered the
the FLUPATH consortium was established. management of some of the recent crises,
resulting in millions of dead animals, con-
FLUPATH is composed of 12 partners, six of cern over loss of human lives and manage-
which are National Reference Laboratories ment of the viruss pandemic potential. For
for avian influenza. The consortium further this reason, and for the devastating effects
includes five academic institutions and two on the poultry industry, international organ-
institutions that specialise in animal science isations such as WHO, OIE and FAO have
and health. The participants, with expertise worked together and established a coord
in chicken genomics, microarray technology, inated set of guidelines and action plans to
pathology, receptors, innate immunity and combat the ongoing Asian epidemic.
Due to the low profile of avian influenza until patterns of its evolution, and its behaviour
1997, a significant amount of information in avian and mammalian species. Work on
and the specific tools necessary to man- currently circulating viruses will allow us
age avian influenza epidemics adequately to track changes in the present situation
are lacking. This includes both the EU situ- and thus issue precise warnings, should the
ation and the ongoing H5N1 crisis. Recent threat of a pandemic increase.
outbreaks of HPAI have affected avian spe-
cies that are showing a reduced susceptibil- It is anticipated that this approach will
ity to this virus. If HPAI infection of the wild permit a more complete understanding
bird host becomes compatible with normal of the immunological, cell biological and
behavioural patterns and migration, the molecular basis of the threat posed by
result will be the development of an endemic HPAIV such as the current H5N1. Essential
cycle in wild birds. The consequences of such information on critical viral and host fac-
a situation are unpredictable and potentially tors, which determine the transmissibility of
very dangerous. the virus to mammals, will be determined.
This knowledge will clearly be of high value
Retrospective analysis of recent outbreaks when implemented in novel strategies to
has permitted the identification of weak combat avian influenza.
points in the management system that
represent areas of uncertainty in which
improvement is required. Several of these Summary of most important
weak points can be directly linked to our results
lack of fundamental knowledge about the WP1 addressed the issue of pathogen-host
importance of both viral as well as host fac- interactions and virulence determinants at
tors in determining the outcome of infec- the molecular level. Little was know about
tion. Therefore, it is essential to increase our the host-response following infection with
effort to enhance our knowledge about the viruses that differ in virulence and gene
ecology and pathology of avian influenza constellation. The contribution of specific
virus infections in poultry and other species. viral genes or gene sequences to
pathogenesis, host- and tissue-tropism as
well as their role in interference with host
Aim defence mechanisms have been examined
This proposal aimed to generate data on in vivo in different species in animal experi-
significant issues linked to AI outbreak ments as well as in vitro. Gene-expres-
management on which scientific knowledge sion profiling and suppression subtractive
is currently lacking. These issues are all hybridisation (SSH) were used to identify
related to the virus-host interactions. Four host genes specifically involved in the reac-
major tasks (work packages WP) were tion to infection by different avian influ-
identified. WP1 studied the host-pathogen enza virus genotypes. The involvement and
interactions. WP2 was dedicated to the importance of viral genes as well as host
ecology and pathogenesis of the viruses. genes were evaluated by cross-validation
WP3 focused on receptor specificity and and in vitro experiments.
interspecies transmission whereas WP4
was concerned with the immunology/innate Chicken and ducks were infected either
immunity response in the infected host. with highly pathogenic avian influenza
virus (HPAIV) strain H7N1 or a low patho-
genic avian influenza virus (LPAIV) strain
Expected results H7N1. LPAIV was able to spread system
The FLUPATH proposal improved our ically in chickens and was even found in
understanding of the origins of HPAIV, the the brain. A cytokine storm was not found
in chickens due to H7 HPAIV infection as Experiments on pigs were also conducted

has been found for H5 HPAIV in some to assess their role in maintenance and
mammalian species. However, the cytokine transmission of avian influenza since pigs
expression was only investigated within may serve as a mixing vessel for avian
the first 24 hours post-infection in the and human viruses. The results demon-
lungs. Analysis of host responses revealed strated that an avian influenza virus has the
that chicken and ducks respond differently capacity to infect pigs under experimental
on infection with LPAIV. The virus prefer- conditions but it is not sufficient to allow
entially replicated in respiratory tissue in transmission to another pig or ferret (model
chickens and in intestinal tissue in ducks. for humans). Such transmission events are
A notable difference was observed in the thought to be involved in the adaptation
interferon response between ducks and of an avian influenza virus to a mamma-
chickens in different organs. Whether, and lian host. The replication level of an avian
how, the difference in these responses influenza virus in pigs is not necessarily a
is also correlated with the difference in good parameter for its capacity to transmit
clinical outcome is not known. Finally, our to another mammalian host. Because the
results confirm the notion that differences replication of both swine and avian influ-
in the outcome of influenza virus infec- enza viruses reached the highest levels in
tion are determined by multiple genetic the lungs, future studies will have to be
determinants. performed to assess if there is a correlation
with the generation of reassortants in this
WP2 studied the ecology and pathology of location.
different avian influenza strains in different
host species. Specific attention was given to The NS1 protein is a multifunctional protein
waterbirds such as ducks which seem to be in AIV that counteracts the host immune
clinically less susceptible to highly patho- response by blocking the synthesis of type
genic avian influenza and, therefore, are I interferon (IFN). Recent work has identified
implicated in transmission of the virus. The truncations and elongations of the C-ter
Asian lineage H5N1 (HPAIV) caused large minus of the NS1 protein of a number of
numbers of deaths in both poultry and wild different avian influenza isolates. The aim
bird populations. Recent isolates of this here was to determine what role these
virus have been reported to cause disease alterations of the C-terminus could play in
and death in commercial ducks, which has the pathogenicity of the viruses that pos-
only rarely been reported for other HPAI sess them. The analysis has clearly shown
viruses. Although the classic dogma main- that the length variations seen between
tains that HPAI virus infection in duck and influenza A viruses isolated from different
waterfowl is limited to the respiratory and species is not a random event. The removal
enteric tract, the results showed that three or addition of specific amino acid residues at
of the six classic HPAI viruses are able to the C-terminal of the protein must confer an
replicate in locations different to the res- advantage or disadvantage on the viruses.
piratory and gastrointestinal tract, which The effects on the host immune system of
can lead to increased lethality for the host. viruses carrying different NS1 truncations
The identification of genetic markers that was also investigated (see WP4).
determine the pathogenicity of HPAI for
ducks requires further characterisation, WP3 tested the role of receptor specificity
coupled with pathogenesis studies aimed and neuraminidase activity of avian viruses
at the understanding of the innate immune in interspecies transmission, pathogenicity
response in varying age ducks that allows and the emergence of potentially new pan-
them to modulate the outcome of HPAI demic strains. We characterised the recep-
infection. tor specificity of more than 200viruses
from different avian species. All H7 sub- epithelium. These findings highlight the
type viruses, irrespective of host species, importance of further systematic studies
displayed a poultry-virus-like binding. This in order to identify avian species and viral
may explain the propensity of aquatic bird binding phenotypes with a high potential for
H7 viruses to cause outbreaks in poultry. zoonotic transmission to humans.
Many also showed weak but significant
binding to human-type receptors, consist- In WP4, gene expression analyses revealed
ent with the ability of H7 viruses to cause that H5N1 disseminated to multiple organs
occasional human infections. where immune responses could be iden-
tified. Among those cytokines strongly
Different receptor phenotypes might pro- induced following H5N1 infection were the
vide the virus with an enhanced potential Th1-associated cytokines but not Th2-ass
for interspecies transmission to pigs and ociated cytokines. The role of the HA pro-
humans. Molecular mechanisms for avian- tein in eliciting in vivo and in vitro cytokine
to-avian, avian-to-pig and avian-to-human responses was also investigated. The data
transmission were studied by using dif- obtained support the cytokine storm
ferent cells and tissues including tracheal hypothesis to explain the particular viru-
explants from chickens, turkeys and ducks, lence of HPAI H5N1 in chicken, placing typeI
and cultures of human airway epithelium. IFN and Th1 cytokine responses at the cen-
In ex vivo porcine organ cultures, we could tre of the immunopathological events.
show a clear, but not absolute, correlation
between the receptor distribution and the A method to generate chicken bone mar-
sensitivity of these tissues to infection with row-derived dendritic cells (DC) was devel-
a particular virus. Our findings indicate that oped. While infection of chicken DC with
changes in the binding tropism towards LPAIV did not affect their activation, HPAIV
a more avian-like virus reduce (but do induced enhanced DC function. Both LPAIV
not abolish) the virus replication in nasal, and HPAIV possess the ability to induce
tracheal and especially bronchial organ type I IFN in chicken leukocytes, implying
cultures. These results, for the first time, the presence of a plasmacytoid-like DC
directly demonstrated that receptor speci- population, which efficiently responds to
ficity of influenza viruses contribute to their AIV despite the presence of the IFN antago-
distinctive tissue tropism in swine respira- nist NS1. Altogether, this knowledge on the
tory tract. Our results also indicate that an early immune response against infection
exclusive change of the binding tropism of with avian influenza viruses may be very
a virus is not sufficient to change its host important for the development of vaccines.
tropism. Therefore, pandemic risk assess- Targeting the early innate immune response
ments of newly isolated viruses in the field, which starts within hours after vaccination
should not only be based on a change of may be a very effective way of controlling
binding tropism. Finally, our results suggest virus replication, thereby limiting tissue
that the susceptibility of pigs to avian influ- damage and promoting adaptive immune
enza viruses was overestimated in the past responses required for protective immunity.
(and therefore also its exclusive role as a
mixing vessel for the generation of new Human pulmonary microvascular endothe-
pandemic viruses). lial cells had a clearly higher susceptibil-
ity to infection by H5N1 HPAIV than to
Studies of different H5N1 showed marginal infection by human influenza viruses. This
but statistically significant differences dem- was related to a relatively higher binding
onstrating that fine distinctions in the recep- capacity to cellular receptors and associ-
tor specificity of avian viruses may affect ated with endothelial cell activation and
viral replication efficiency in human airway apoptosis. Reverse genetics analyses
demonstrated a major role for HA in this type I responses in the lung, plasma and
cell tropism. Overall, avian H5N1 viruses spleen during highly pathogenic H5N1
have a particular receptor specificity tar- infection of chicken, Veterinary Research,
geting endothelial cells that is different 42: 6.
from human influenza viruses. This H5N1
receptor specificity could contribute to Rebel, J.M.J., Peeters, B., Fijten, H., Post, J.,
disease pathogenesis. Cornelissen, J., Hoek, A., Vervelde, L. (2011),
Highly pathogenic or low pathogenic avian
Investigations on NS1 revealed that specific influenza virus subtype H7N1 infection in
C-terminal truncations are shared between chicken lungs: small differences in gen-
different influenza A subtypes indicating a eral acute responses, Veterinary Research,
potential role in influenza infectivity and 42:10.
pathogenicity. Additional analysis of recom-
binant viruses also revealed a potential role Van Poucke, S., Nicholls, J., Nauwynck,
of these truncations in altering the expres- H., Van Reeth, K. (2010), Replication of
sion of iNOS with an associated reduction in avian, human and swine influenza viruses
NO production. in porcine respiratory explants and asso-
ciation with sialic acid distribution, Virology
Journal, 7: 38.
FLUPATH will ultimately provide knowledge
and tools for new strategies which will be Project website
tailored for the control and management of http://www.flupath.eu
AI at both European and international level.
This should result in a noticeable reduction Keywords
in the impact that this disease has had in avian influenza virus, virus-host inter
the past. actions, virus-receptor interactions, host-
specificity, expression profiling, virulence,
pathology, ecology, innate immunity
Giannecchini, S., Clausi, V., Di Trani, L., Fal- Coordinator
cone, E., Terregino, C., Toffan, A., Cilloni, F., Dr Ben Peeters
Matrosovich, M., Gambaryan, A.S., Bovin, Central Veterinary Institute of Wageningen
N.V., Delogu, M., Capua, I., Donatelli, I., University and Research Centre
Azzi, A. (2010), Molecular adaptation of Houtribweg 39
an H7N3 wild duck influenza virus follow- 8203 AA Lelystad
ing experimental multiple passages in quail NETHERLANDS
and turkey, Virology, 408: 16773. Tel. +31 320238693
ben.peeters@wur.nl
Jansen, C.A., van de Haar, P.M., van Haarlem,
D., van Kooten, P., de Wit, S., van Eden, W.,
Viertlbck, B., Gbel, T.W.G. and Vervelde, L.
(2010), Identification of new populations of
chicken natural killer (NK) cells, Develop-
mental and Comparative Immunology, 34:
759767.
Moulin, H.R., Liniger, M., Python, S., Guzy-

lack-Piriou, L., Ocana-Macchi, M., Ruggli,
N., Summerfield, A. (2011), High interferon
Partners
ORGANISATION CONTACT NAME CONTACT
Agence Franaise de Scurit Dr Vronique Jestin v.jestin@ploufragan.afssa.fr
Sanitaire des Aliments
AFSSA site de Ploufragan
B.P. 53 22440 Ploufragan
FRANCE
CSIRO Livestock Industries, Dr John Lowenthal, john.lowenthal@csiro.au
Australian Animal Health Dr Ton Schat kas24@cornell.edu
Laboratory
Post Bag 24
5 Portarlington Rd
3220 Geelong
AUSTRALIA
Istituto Zooprofilattico Speri- Dr William Dundon, wdundon@izsvenezie.it
mentale delle Venezie Dr Ilaria Capua icapua@izsvenezie.it
Laboratorio di Virologia
Viale dellUniversit 10
35020 Legnaro (PD)
ITALY
University of Ghent Dr Kristien van Reeth, Dr kristien.vanreeth@UGent.be
Faculty of Veterinary Medicine Karen van de Meulen karen.vandermeulen@ugent.be
Salisburylaan 133
9820 Merelbeke
BELGIUM
Veterinary Laboratories Agency Dr Jill Banks i.h.brown@vla.defra.gsi.gov.uk
Woodham Lane j.banks@vla.defra.gsi.gov.uk
Addlestone
Surrey KT15 3NB
UNITED KINGDOM
Institute of Virology Dr Mikhail Matrosovich m.matrosovich@gmail.com
Philipps University
Hans-Meerwein-Str. 2
35043 Marburg
GERMANY
Tierrztliche Hochschule Dr Georg Herrler georg.herrler@tiho-hannover.de
Hannover
Institute of Virology
Bnteweg 17
30559 Hannover
GERMANY
Institute of Virology and Dr Artur Summerfield, Dr artur.summerfield@ivi.admin.ch
Immunoprophylaxis Ken McCullough ken.mccullough@ivi.admin.ch
3147 Mittelhusern
SWITZERLAND
Utrecht University Dr Willem van Eden, Dr w.eden@vet.uu.nl
Faculty of Veterinary Medicine Lonneke Vervelde l.vervelde@vet.uu.nl
Division of Immunology
Yalelaan 1
3508 Utrecht
NETHERLANDS
ORGANISATION CONTACT NAME CONTACT

The Roslin Institute, University Dr David Burt dave.burt@roslin.ed.ac.uk
of Edinburgh
Division of Genetics and
Genomics
Roslin Institute
Midlothian, EH25 9PS
UNITED KINGDOM
The Hebrew University of Dr Dan Heller dheller@agri.huji.ac.il
Jerusalem
Faculty of Agricultural Food and
Environmental Sciences
Dept of Animal Sciences
PO Box 12
Rehovot 76100
ISRAEL
[New-Flubird]
Network for early warning

of influenza viruses in
migratory birds in Europe
Summary AIVs in different scenarios, on a scale and
Acronym:
New-Flubird
The overall objective of New-Flubird depth previously not possible. The future
to provide Europe with an early warn- challenge is to maintain a baseline sam-
Project number:
ing and risk assessment system for the ple flow into the New-Flubird database,
044490
threat posed to animal and human health whereby targeting certain indicator spe-
EC contribution: by avian influenza (AI) viruses in migra- cies in defined high-risk regions and pos-
EUR 1 855 350
tory birds has been achieved. With sibly at selected times during the year
Duration: the identification of higher risk species where appropriate capacity and expertise
36 months by experimental infection in the labora- will also need to be assured.
Start date: tory, the development of flyway maps
1 January 2007 and associated identification of high-risk
Instrument: sites, the upgraded skills and expertise Problem
Specific Targeted levels, the establishment of a sampling The threat posed to animal and human
Research or network, the integration of the data and health by avian influenza viruses from
Innovation Project development of mathematical models, a migratory birds.
(STREP)
firm basis has been obtained for AI virus
(AIV) surveillance in migratory birds. The
available data, assembled in the New- Aim
Flubird database and the models devel- The urgent development of early warn-
oped allow for investigating the spread of ing and rapid response systems to combat
animal and human influenza threats posed help people easily obtain information on the
by AI viruses from migratory birds. most important sites for migratory water-
birds at both national and international
level.
Results
The waterbird migration in and to Europe was At the basis of the New-Flubird activities
described in maps to facilitate risk assess- and results lies the creation and mainten
ment and selection of sites for AI sampling ance of a network of sampling sites for
in wild birds. Furthermore, a list of higher sampling AIVs in wild birds as a component
risk species was developed: a review of the of an early warning system. Concerning the
current situation of highly pathogenic avian mortality monitoring, New-Flubird decided,
influenza (HPAI) H5N1 in and around Europe after consultation with a selected group of
was initiated, leading to a preliminary list in advisors coordinating waterbird counting in
2007 and resulting in a finalised list, which the framework of the International Waterbird
all ornithological partners within the New- Census (Wetlands International), to recon-
Flubird consortium finalised in 2008. A list of sider its efforts in relation to the develop-
82 higher risk species was issued and inte- ment of a mortality-monitoring programme
grated into the New-Flubird database with because this activity was covered by others.
the technical tools (coding systems) required. Concerning live bird monitoring, New-Flubird
has collected over 25 000 samples in over 10
Based on data from the International countries across Europe and Africa during the
Waterbird Census (IWC, Wetlands Inter course of New-Flubird, covering the higher
national) and the Important Bird Areas risk species sites. The sampling effort was
database (Birdlife International), New- accompanied by a range of on-site capacity-
Flubird established and published combined building activities in wild bird capturing and
migratory flyways allowing for the selection sampling (e.g. duck traps, duck decoys, swan
of important sites for inclusion in the early pipes and cannon netting, sample collection,
warning system network and for forecast- storage and transport, personnel health and
ing and modelling movements of birds in safety and animal welfare).
the event of an outbreak. All flyway maps
were finalised and made available through The collected samples were screened for
GIS shape files to be introduced in the the presence of influenza A virus, the influ-
New-Flubird database and published in the enza viruses subtypes characterised by hae-
Wader Atlas in 2009. magglutination inhibition (HI) assays and/
or nucleotide sequence analysis. The infor-
Also based on the IWC, the high numbers mation on the prevalence of low pathogenic
of higher risk species counts per site were avian influenza (LPAI) and HPAI viruses in
mapped for each species and then com- wild birds in Europe was integrated into the
bined. This analysis allowed the identifica- New-Flubird database, together with over
tion and the targeting of sites for strategic 145 000 samples from the active and pas-
surveillance and for establishing a network sive monitoring of the EU Member States.
of semi-continuous sites.
New-Flubird contributed to the standard
The Wader Atlas was officially launched dur- isation of field and laboratory methodology
ing the African-Eurasian Waterbird Agree- and training of the technical and scientific
ments 15th Anniversary in The Hague, personnel in three consortium workshops,
the Netherlands, providing Europe with establishing and validating methods that
comprehensive information on 561 popu- are robust, repeatable and safe for person-
lations and 294 waterbird species from 3 nel and wildlife, thereby ensuring a highly
020 wetland sites. It has been designed to qualitative process from sample in the field
to result in the participating laboratories. diving duck species. The main clinical signs
In addition, New-Flubird has built capacity were neurological and were caused by viral
for influenza surveillance in wild birds by encephalitis. In contrast, the remaining four
organising training courses and workshops species of dabbling ducks were clinically
at key regions in Africa and Europe on the unaffected. Pharyngeal excretion of HPAIV
crucial skills sets for safe and effective wild (H5N1) varied significantly among the six
bird surveillance. duck species and cloacal excretion was
uncommon. Of the six wild duck species
A large set of representative virus isolates studied, the mallard (Anas platyrhynchos)
has been characterised genetically by is the prime candidate for being a long-
full genome sequencing. This set of virus distance vector of HPAIV (H5N1) because
genome sequences serves as a reference it was the only species to show abundant
set in the public sequence databases that virus excretion without clinical or pathologic
are used by the scientific community. evidence of debilitating disease. Pochards
and tufted ducks are more likely to act as
sentinels for HPAIV (H5N1) in wild bird pop-
ulations. However, pochards cannot be ruled
out as potential vectors because some
birds excreted abundant virus in absence of
severe clinical signs.
The stress hormone corticosterone contrib-

utes to regulating physiological changes that
prepare wild birds for migration, but also
may lead to higher susceptibility of migra-
tory birds to infection. The effect of these
physiological changes on the spread of HPAIV
H5N1 in migratory birds is unclear. Therefore,
we infected red knots (Calidris canutus) with
HPAIV H5N1 before the migration period, just
prior to migration, at the time of migration,
Next to the surveillance and capacity-build- and after the migration period and measured
ing activities, New-Flubird has also focused both clinical signs and virus excretion. Red
on identifying migratory and other wild knots infected at the time of migration had
bird species that could act as spreaders pharyngeal virus titres that were strongly
or sentinels for HPAI (H5N1) virus infec- positively correlated with plasma concentra-
tion and the assessment of the contribution tion of corticosterone. Furthermore, birds that
of contacts with migratory and other wild developed severe disease had higher pharyn-
bird species to the overall risk posed by this geal virus titres than those that remained
virus infection to poultry, other birds, and asymptomatic. Therefore, wild birds in migra-
mammals (including humans). To test the tory condition may shed more HPAIV H5N1
hypothesis that wild waterbirds can excrete and may be more susceptible to developing
HPAIV (H5N1) in the absence of debilitat- severe disease, which may shorten or delay
ing disease and so potentially act as long- migratory flights, slowing yet not necessarily
distance virus vectors, groups of six species abrogating the geographical spread of HPAIV
of wild ducks were infected experimentally H5N1 by wild migratory birds.
with HPAIV (H5N1). HPAIV (H5N1) infec-
tion caused clinical signs of disease only in At the heart of New-Flubird was the estab-
tufted ducks (Aythya fuligula) and common lishment of a web-based database manag-
pochards (Aythya farina), both of which are ing surveillance data of AIVs in wild birds
as a central data repository describing and conducted within the framework of the
analysing the epidemiology of AIV infection New-Flubird project.
in wild bird populations.
Characteristic migration patterns for dif-
For this purpose, laboratory data are com- ferent species were also included in the
bined with ornithological and environmental model.
data. As a central instrument, a database
system has been developed to store, man- Burn-in runs highlighted parameters to
age and analyse data from the different which the model reacted sensitively. By
disciplines. Flanking environmental data defining rule sets in accordance with H5N1
are provided as well. Data access rights outbreaks observed in the wild (e.g. regard-
can be configured independently for differ- ing observed lengths of outbreaks and
ent users, and different data types, respec- measured prevalences), thresholds were
tively. Interaction by project participants is defined for these. Changes in the overall
possible via a secured Internet connection length of the epidemic course, as well as
and a web interface, which provides the dif- amplitude and time point of the number
ferent tools and modules for data process- of maximally infected birds were used as
ing. An exchange of data and information readouts.
with related initiatives, for example the wild
bird monitoring campaigns in the European New-Flubird has achieved the vast major-
Union or the Global Avian Influenza Network ity of its planned results. In some cases,
for Surveillance (GAINS) of the World Con- New-Flubird has gone beyond its original
servation Society (WCS), is easily achieved plans (e.g. the New-Flubird database con-
since data structures and coding systems tains data from the International Waterbird
were implemented and designed to preserve Census, with millions of records, to provide
compatibility. Emphasis is placed on the background on waterbird distribution with
integrative process of combining the inter- time series) whilst in other areas, it did not
disciplinary data for analysis, which is real- meet expectations. This includes monitoring
ised on different levels. Interactive software the occurrence of dead birds in the field as
modules allow for the creation of database part of the International Waterbird Census,
queries and target parameters which are which has not proven feasible and another
shared by the different types of data. Basic approach will be needed. The overall objec-
and advanced features of the New-Flubird tive of New-Flubird to provide Europe
database have been introduced to New- with an early warning and risk assessment
Flubird partners in two workshops. system for the threat posed to animal and
human health by avian influenza (AI) viruses
New-Flubird has developed an agent- in migratory birds has been achieved.
based, stochastic epidemiological model
assuming a three-species scenario with
intra- and interspecies transmissions in Potential applications
mallards, mute swans (Cygnus olor) and The knowledge gained from the experimen-
common pochards at fixed geographic tal infection studies has several implica-
localities and during migration. For all spe- tions for surveillance in wild ducks. Active
cies, juvenile animals were modelled to surveillance should give priority to mallards
have a twofold higher susceptibility com- and, to a lesser degree, pochards. Sampling
pared to adults. Course and outcome of should not be limited to cloacal swabs, but
HPAIV H5N1 infection for the individual should include pharyngeal swabs. Passive
bird was modelled along species-depend- surveillance should pay extra attention to
ent characteristics based on findings from tufted ducks and pochards. Sampling of
experimental infection studies partly wild duck carcasses should not be limited
to cloacal, pharyngeal, and tracheal swabs techniques such as blood sampling, feather
and should include internal organs. collection and water sampling can be imple-
mented in the capacity-building activities.
The developed agent-based, stochastic epi-
demiological model allows for investigat-
ing the spread of HPAIV H5N1 in various References/publications
scenarios. Starting with a single-location Artois, M., Bicout, D., Doctrinal, D., Fouchier,
scenario, each modelling step, correspond- R., Gavier-Widen, D., Globig, A., Hagemeijer,
ing to one day, simulates randomised con- W., Mundkur, T., Munster, V., Olsen, B., Out-
tacts between individuals of the different breaks of highly pathogenic avian influenza
model compartments: Susceptible, Exposed, in Europe: the risks associated with wild
Infectious, Removed (SEIR). By varying the birds, Rev. Sci. Tech., April 2009, 28(1):
effective contact rates within and between 6992.
model species, differences in feeding habits
and seasonal gregariousness are respected. Hoye, B.J., Munster, V.J., Nishiura, H.,
Klaassen, M., Fouchier, R.A.M., Surveil-
When considered in relation to the onset of lance of wild birds for avian influenza
migration, the chance of migration-capable virus, Emerging Infectious Diseases, 16:
birds propagating the virus and spread- 182734(2010).
ing over larger distances can be exam-
ined. Furthermore, the current surveillance Keawcharoen, J., van Riel, D., van Amer-
schemes in wild birds using the model and ongen, G., Bestebroer, T., Beyer, W.E., van
data from the New-Flubird database were Lavieren, R., Osterhaus, A.D., Fouchier, R.A.,
re-evaluated. Kuiken, T. (2008), Wild Ducks as Long-Dis-
tance Vectors of Highly Pathogenic Avian
With the identification of higher risk species, Influenza Virus (H5N1), Emerging Infectious
the development of flyway maps and asso- Diseases, 14: 600607.
ciated identification of high risk sites, the
improved capacity and expertise levels, and Munster, V.J, Baas, C., Lexmond, P., Wal-
the establishment of a sampling network, denstrom, J., Wallensten, A., Fransson, T.,
a firm basis has been obtained for the AIV Rimmelzwaan, G.F., Beyer, W.E.P., Schutten,
surveillance in migratory birds. The available M., Olsen, B., Osterhaus, A.D.M.E., Fouchier,
data, assembled in the New-Flubird data- R.A.M. (2007), Spatial, temporal and species
base and the models developed allow the variation in prevalence of influenza A virus in
investigation of the spread of AIV in different wild migratory birds, PLoS Pathogens, 3:e61.
scenarios, on a scale and depth previously
not possible. The future challenge is to main- Roche, B., Lebarbenchon, C., Gauthier-
tain a baseline sample flow into the New- Clerc, M., Chang, C-M., Thomas, F., Renaud,
Flubird database, focusing on certain indica- F., van der Werf, S., Gugan, J.-F. (2009),
tor species in certain high risk regions Avian influenza dynamics in wild birds are
and possibly (at certain times) target species driven by water-borne transmission, Infec-
at key sites where appropriate capacity and tion Genetic and Evolution, 9 (5): 800805.
expertise will also need to be assured.
Project website
If this network is extended and strength- http://www.new-flubird.eu
ened, geographic gaps in the surveillance
(Caspian Sea, Lake Chad and other areas) Keywords
can be covered, surveillance of dead birds avian influenza, virus infections, flyway,
can be implemented, existing pathology database, early warning system, migratory
networks can be involved and other new birds, poultry, H5N1, highly pathogenic
Coordinator Nicolas Tubbs

Prof. Dr A.D.M.E. Osterhaus DVM, PhD Wetlands International
Erasmus MC Biodiversity & Ecological Networks
s-Gravendijkwal 230 P.O. Box 471
3015 CE Rotterdam 6700 AL Wageningen
NETHERLANDS NETHERLANDS
a.osterhaus@erasmusmc.nl Tel. +31 318660927
Nicolas.tubbs@wetlands.org
Co-Coordinator
Ward Hagemeijer
Wetlands International
Biodiversity & Ecological Networks
P.O. Box 471
6700 AL Wageningen
NETHERLANDS
Tel. +31 318660910
Ward.Hagemeijer@wetlands.org
Partners
Friedrich-Loeffler-Institut Dr C. Staubach timm.harder@fli.bund.de
Institut fuer Epidemiologie, Institut Prof. Dr T C Harder
fuer Virusdiagnostik
Sdufer 10
17493 Greifswald-Insel Riems
GERMANY
Linnaeus University Prof. B. Olsen bjorn.olsen@medci.uu.se
Section for Zoonotic Ecology and
Epidemiology
Dept of Biology and Environmental
Science
SE-391 82 Kalmar
SWEDEN
Danish Institute for Food and Dr P.H. Jorgensen pojo@vet.dtu.dk
Veterinary Research
Hangvej 2
8200 Aarhus N
DENMARK
National Veterinary Research Dr Zenon Minta zminta@piwet.pulawy.pl
Institute
Department of Poultry Diseases
Al Partyzantow 57
24-100 Pulawy
POLAND
National Veterinary Institute Monika Hjortaas monika.hjortaas@vetinst.no
Section for Virology and Serology
P.O. Box 750 Sentrum
N-0106 Oslo
NORWAY

Istituto Zooprofilattico Sperimentale Dr C. Terregino cterregino@izsvenezie.it
delle Venezie
Viale dellUniversit 10
35020 Legnaro (PD)
ITALY
Foundation Tour du Valat Dr M. Gauthier-Clerc@tourduvalat.org
Le Sambuc Gauthier-Clerc
13200 Arles
FRANCE
Veterinary Laboratories Agency Dr I. Brown i.h.brown@vla.defra.gsi.gov.uk
Woodham Lane
New Haw
Addlestone, KT15 3NB
Surrey
UNITED KINGDOM
UR Gestion Intgre de la Faune Dr F. Monicat francois.monicat@cirad.fr
Avenue Agropolis
34398 Montpellier Cedex 5
FRANCE
Wildfowl & Wetland Trust Dr R. Cromie Ruth.Cromie@wwt.org.uk
Slimbridge, GL2 7BT
Gloucestershire
UNITED KINGDOM
OMPO Dr Alexandre vanneau@ompo.org
5 Ave des Chasseurs Czajkowski
75017 Paris
FRANCE
[FLURESIST]
Avian influenza virus survival

in poultry commodities,
poultry manure and the
environment
Summary knowledge about virus content of com-
Acronym:
Avian influenza (AI) outbreaks have recently modities, the stability of the virus in these FLURESIST
caused severe losses to the poultry indus- products, in litter and the environment, is
Project number:
try, its stakeholders and, ultimately, to the needed.
044311
EU taxpayer. In addition, the ongoing Asian
H5N1 outbreak is a serious concern for food EC contribution:
EUR 870 000
security and human health worldwide. Problem
The circulation of the HPAI virus in Asia and Duration:
36 months
In Asia, due to both social conditions and in the Middle East and Africa could rep-
the particular characteristics of the H5N1 resent the origin of a pandemic virus for Start date:
virus, the crossing of the species barrier humans, and many questions have been 1 March 2007
represents a serious potential risk of a new raised with a view to finding a way to com- Instrument:
human pandemic virus emerging. Evidence bat the ongoing AI crisis. Due to the lack of Specific Targeted
is growing that highly pathogenic avian field and experimental data, certain ques- Research or
Innovation Project
influenza (HPAI) H5N1 is not only spread- tions on virus survival in the environment
(STREP)
ing through trade but is also carried by and in poultry and other avian commodities
wild birds. H5N1-infected wild birds, mainly are not yet answered, and these knowledge
waterfowl, have been detected in the EU gaps should be filled following the results
in Germany, Greece, France, Italy, Austria, of the ongoing and new research efforts of
Poland and Sweden. These findings raise the scientific community.
our awareness that H5N1 is becoming
ever more endemic in wild birds. The dis-
covery of infected mammals such as cats, Aim
leopards, stone martens and raptors that The aim of the project is to obtain data
died as a result of infection with H5N1 and provide knowledge on the presence of
has uncovered the consequences of this influenza viruses in commodities and lit-
development. ter of infected poultry. In order to develop
validated protocols for cleansing, disin-
Questions are being raised about the risk fection and treatment of litter, and to be
of contamination of surface water in rela- able to assess the risk of carcass disposal,
tion to the health of other animals and treated litter and poultry commodities
humans. To answer these questions and to such as meat, feathers and eggs, virus
be able to assess the risks involved in trad- survival will be determined in a standard-
ing in poultry commodities and litter, more ised manner in different environments.
The project also aims to create knowledge severe consequences for those engaged in
about environmental factors that influence the industry. Similarly, consumer misgiv-
virus stability. Data collected will be used ings over the safety of poultry, and even
for proper risk assessment of the trade meat or eggs of vaccinated poultry, have
in treated and fresh poultry commodi- to be addressed even for consumption at
ties, poultry litter and the contaminated home. Finally, in the event of an outbreak,
environment. there would be a need to dispose of a large
quantity of carcasses and litter and the
virological consequences of this have to
Expected results be considered if we are to avoid potential
Research will provide data on the survival contamination problems for ground and
of different AI viruses and the effect of surface water, and even possibilities of
physical parameters such as pH and tem- virus survival in soil. For these reasons, this
perature on survival. project is designed to provide data on the
levels and stability of the virus in differ-
Virus concentrations in poultry commod ent materials, particularly avian materials
ities such as meat, feathers and eggs will and products, and thus help calm public
be determined. Studies will generate quan- concerns.
titative data on survival of viruses in such
commodities at ambient temperatures
and at temperatures used to treat poultry Project website
products. http://www.fluresist.eu
Knowledge will be obtained on whether Keywords

certain soils, lake silts and living organisms, avian influenza, poultry commodities, virus
enteric factors of waterfowl, sterile faeces survival, biosecurity measures, disinfection,
or gut flora and sewage pollution increases cleansing operations, public health risk
or decreases virus survival. Based on the
results, protocols for waste treatment, Coordinator
carcass disposal and disinfection will be Dr Guus Koch
adapted and/or validated. Data will be used Centraal Veterinary Instituut Wageningen
to make proper risk assessments. University Research
P.O. Box 65
Houtribweg 39
Potential applications 8200 AB Lelystad
The current threat from AI poses serious NETHERLANDS
threats to the poultry industry, and perhaps Tel. +31 320238800
eventually to man. In view of these threats, Guus.Koch@wur.nl
society faces a number of problems and
this project is designed to provide the scien-
tific data to underpin the formulation of any
response and a basis for the development
of further biosecurity measures. Any such
response must be driven by an understand-
ing of the behaviour of the virus. Threats
to industry include the introduction of the
virus from wild populations or trade; this,
in turn, threatens the competitiveness of
the industry: an outbreak would certainly
restrict a nations trading capacity and have
Partners
ORGANISATION CONTACT
Istituto Zooprofilattico Sperimentale delle Venezie Dr Ilaria Capua DVM
Legnaro
ITALY
Veterinary Laboratories Agency Dr Ian Brown
Addlestone
UNITED KINGDOM
Kimron Veterinary Institute (KVI) Dr Irit Davidson
Bet Dagan
ISRAEL
Sttens Veterinarmedicinska Anstalt (SVA) Dr Jakob Ottoson
Uppsala
SWEDEN
University of Surrey Dr Michael J. Carter
Guildford
UNITED KINGDOM
In collaboration with the project

ORGANISATION CONTACT
US Department of Agriculture Dr D.E. Swayne
Athens
UNITED STATES
Canadian Food Inspection Agency (CFIA) Dr Loyd Spencer
Ottawa
CANADA
US Department of Agriculture Dennis A. Senne
Ames
UNITED STATES
[Rivers]
Resistance of influenza
viruses in environmental
reservoirs and systems
Summary of virus survival from a virological view-
Acronym:
Rivers
The surge of the global avian influenza (AI) point; (b) understand the impact of physical
epizootic mainly caused by the genotype and chemical elements on virus survival; (c)
Project number:
Z highly pathogenic avian influenza virus evaluate the role of environmental reser-
044405
(HPAIV) has posed numerous questions, in voirs; (d) propose standardised protocols
EC contribution: particular to risk managers and policymak- for the concentration and detection of AIVs
EUR 1 395 000
ers. Scientific knowledge is scarce on many in waters, including waste waters, and in
Duration: aspects of the ecology and environmental different matrices including food; and (e)
36 months properties of HPAIVs, in particular H5N1. provide a database together with analytical
Start date: Virus survival, a key element in control tools to allow the generation of evidence-
1 February 2005 strategies, is an illustration of this paucity based guidelines for the prevention and
Instrument: of knowledge. Data from the literature on control of influenza outbreaks in animal and
Specific Targeted AIV survival is rather limited, often very old human populations, especially at times of
Research or and sometimes not confirmed from one restocking.
Innovation Project study to another or is even contradictory.
(STREP)
The results obtained with various subtypes
of influenza A viruses cannot be extrapo- Problem
lated to the current A (H5N1) viruses with- Highly pathogenic avian influenza (HPAI)
out careful consideration. Furthermore, epizootics associated with zoonotic human
little information is provided regarding cases.
the survival of IVs in the air and on sur-
faces no standardised protocols exist
to detect AIVs in waters, in the air or in/on Aim
solid matrices. Ideally, the virus detection The overall aim of this project is the pre-
technique to be used should be sensitive, vention and control of AI type A (H5N1) in
quantitative, rapid and routinely applica- the animal population through the following
ble before or after a standardised sam- specific objectives: (a) gathering data on the
pling method, that does or does not include survival of AI viruses (AIVs) in natural envir
concentration. onments; (b) generating scientific know
ledge about the survival of AIVs in experi-
For this project, nine institutions directly mental settings; (c) providing figures about
involved in AIV, three from Asian countries, the effect of various treatments, either
have joined forces in order to investigate chemical (e.g. disinfectants) or physical
the prevention and control of influenza out- (e.g. UV light), on influenza virus survival;
breaks in the animal population at present (d) providing figures on the effect of various
and at the time of restocking. More specific types of food processing on influenza virus
objectives are to: (a) understand the basis survival; and (e) elaborating models about
the survival of AIVs in natural environ- with a pandemic potential in Europe

ments to demonstrate, in connection with and the rest of the world will be drawn
projects relevant to tasks 3 and 4 of the from the data obtained through Riv-
SSP-5B-Influenza call, their perpetuation in ers in afinal report to the European
nature both in biological and environmental Commission to allow evidence-based
reservoirs. policymaking.
International guidelines for the control
and prevention (through virus inactiva-
Expected results tion and disinfection, for example) of
Criteria for bio-equivalence in relation to outbreaks in domestic birds but also
virus survival between IV strains. in humans will benefit from the data
Method of virus viability assessment generated by the project.
other than virus titration on cell culture,
approvable and standardised protocols
for influenza virus recovery from various Project website
surfaces, approvable and standardised http://www.rivers-project.eu
protocols for testing the effect of chem
ical and physical treatments of different Keywords
types of water on influenza virus sur- influenza, avian, pathogenic, zoonotic,
vival, standard operating procedures for resistance, inactivation, virus, environment,
virus disinfection/inactivation in different reservoirs, systems, models, restocking
settlements.
Data on the prevalence of AIVs in Coordinator
aquatic environments (lakes, ponds and Dr Jean-Claude Manuguerra
rivers) throughout the year and along Institut Pasteur
the stream of rivers, data on gastropods 25 rue du Docteur Roux
and bivalve molluscs regarding their 75724 Paris Cedex 15
potential role as concentrators of AIVs in FRANCE
aquatic biotopes. Tel. +33 14 0613807
Data (database) on IV survival in the air jmanugu@pasteur.fr
and on various kinds of surfaces and
under various conditions, data on the
prevalence of AIVs in the surroundings of
farms with present and past outbreaks
throughout the year.
Descriptive, data-driven, low-level simu-
lation models of AIV perpetuation, viabil-
ity and deactivation in: (a) various water
environments, laboratory-controlled and
natural; (b) in air in laboratory-controlled
environments; (c) in avian faeces and
farm manure.
Multi-scale agent-based simulation
model of possible determinants for AIV
stability, perpetuation and deactivation.
Recommendations for the prevention
and control of current and future AI
outbreaks in wild and domestic birds
Partners
ORGANISATION COUNTRY
Cantacuzino Bucharest, ROMANIA
National Institute of Research and Development Microbiology
and Immunology
The Stephan Angeloff Institute of Microbiology Sofia, BULGARIA
Institut Pasteur du Cambodge Phnom Penh, CAMBODIA
Pasteur Institute of Shanghai Shanghai, CHINA
Centre de coopration internationale en recherche Montpellier, FRANCE
agronomique
Institut Pasteur de Lille Lille, FRANCE
University of Warsaw Warsaw, POLAND
Wuhan Institute of Virology Wuhan, CHINA
[FLUTEST]
Improved diagnosis and early

warning systems for avian
influenza outbreak management
Acronym:
Summary commercially available tests. We examined
FLUTEST The primary goal of this project was the sophisticated laboratory-based methods,
joint development and application of tech- high throughput techniques for molecular
Project number:
nologies to combat avian influenza (AI) and serological testing, penside testing
044429
infections. This goal was pursued through and simplified tests for use in laboratories
EC contribution: the interaction of leading European insti- with limited resources or experience. Efforts
EUR 1 502 880
tutes along with the collaboration of non- were particularly focused on the validation
Duration: EU laboratories experienced in AI outbreak of tests for use on clinical materials derived
48 months control and management. from Anseriformes, other wild bird species
Start date: and selected mammalian species.
1 February 2007 Studies were conducted to establish the
Instrument: effectiveness of the current EU surveillance
Specific Targeted and early warning systems for AI, and then Problem
Research or to develop blueprints for improvements to In recent years, AI outbreaks have caused
Innovation Project these programmes in disease-free periods severe losses to the poultry industry, its
(STREP)
and during outbreaks. The models include stakeholders and the EU taxpayer. The
criteria for harmonised diagnostic tests for ongoing Asian H5N1 outbreak is a serious
on-farm outbreak investigation. concern for food security and human health.
It is estimated that between 2000 and 2007,
A range of diagnostic tools were developed more than 200 million birds died or were
and evaluated alongside a range of culled following infection with influenza
viruses subtypes H5 or H7. Approximately the impact that control measures have on
50 million of these birds were from Europe. these parameters. Time to initial detection
Human infections have also been reported and the time taken to perform tracings of
in several of these outbreaks. In Asia, the potentially infected premises can have a
crossing of the species barrier represents a strong influence. Delays to the initial detec-
serious risk of a new human pandemic virus tion can be exacerbated by problems with
emerging. the differential diagnosis of poultry dis-
eases that present with similar clinical signs
AI is a highly contagious trans-boundary in the early phases of disease.
animal disease. Thus, the prompt identifi-
cation of infected animals is crucial for con- We developed and evaluated a range of novel
trol and eradication. Surveillance must be molecular based tests for the rapid detec-
targeted to appropriate areas and species, tion and differential diagnosis of suspected
and diagnostic tests must be appropriate cases of HP avian influenza. Some offered no
for the setting in which they will be used, improvement over EU-recommended meth-
be properly validated and fit for purpose. ods, for example multiplex PriProET assays,
but others offered promise for pen-side test-
ing. A range of control reagents was also
Aim developed and validated, potentially leading
The project aims were to generate data to better standardisation and harmonisation
on issues linked to AI surveillance and of testing in the EU Member States. A rapid
outbreak diagnosis and management, (compared with sequencing) and sensitive
on which scientific knowledge is lacking. pathotyping method showed notable promise
We also planned to develop and validate and an international patent application has
laboratory tests that can be used as tools been filed on the method.
in early warning systems and surveillance
programmes for AI, in the presence and Domestic ducks play a pivotal role in LP
absence of vaccination. and HP AI epidemiology and tools for detec-
tion of prior infection by antibody detec-
tion were lacking. We developed a range
Results of ELISAs for the detection of antibodies
Mathematical models have been developed against the H5, H7, N2, N3 and N9 sub-
that can serve as a blueprint for the design types. Assays for the neuraminidase anti-
of a surveillance programme for HPAI as bodies are important where vaccination and
well as for LPAI. These models have been the DIVA (differentiation of infected from
parameterised using data from experiments vaccinated animals) principle are applied.
and epidemics. Moreover, a risk assessment We also assessed the performance of four
model has been developed. commercial antibody detection ELISAs
against the gold standard haemaggluti-
FLUTEST addressed issues of molecular nation inhibition (HI) test for poultry sera.
diagnostics, antigen detection and novel The best ELISAs had high sensitivity but
technologies. Focusing efforts on the lower specificity than HI, although all were
development, evaluation, application and > 90 %. An ELISA and sampling regime was
harmonisation of these novel molecular developed using egg yolk as a sample. This
techniques, for detection and differential test has applications for the automated
diagnosis of AI virus infections in domestic high-volume surveillance of layer flocks at
and free-living avian populations. reduced cost to current protocols.
Many factors can influence the size, spread Label-free AI nucleic acid and/or antigen
and duration of HPAI outbreaks, and also detection was a novel approach we pursued
with the aim of using electrochemical Low-pathogenic notifiable Avian Influenza

detection sensor devices. However, the sen- serosurveillance and the risk of infection
sitivity of these devices currently does in poultry A critical review of the Euro-
not improve on the current PCR-based pean Union active surveillance programme
approach, although we demonstrated the (200507), Influenza and other respiratory
potential for this technology in the future. viruses, 4, 9199.
The use of microarray for AI sample sub- Kukol, A., Li, P., Estrela, P., Ko Ferrigno, P.,
typing was successful both in sensitivity Migliorato, P. (2008), Label-free electrical
and specificity and could offer advantages detection of DNA hybridisation for the
over other tests, however the cost of this example of influenza virus gene sequences,
technology is currently prohibitive. Analytical Biochemistry, 374 (2008),
143153.
Potential applications Leijon, M., Ullman, K., Thyselius, S., Zohari,

Early warning and risk assessment sys- S., Pedersen, J.C., Belk, S., Novel real-time
tems will be used by policymakers as input PCR technique for rapid molecular avian
for decisions and control measures for AI influenza pathotyping (submitted to Jour-
viruses coming from migratory birds. nal of Clinical Microbiology).
The data generated will aid decision-mak- van der Goot, J.A., Engel, B., Van de Water,
ers within the European Commission with S.G., Buist, W., De Jong, M.C., Koch, G., van
regard to the prevention and control of Boven, M., Stegeman, A. (2010), Valid
epizootic disease of poultry. The project ation of diagnostic tests for detection of
addresses the needs within the EU for sus- avian influenza in vaccinated chickens using
tained improvements in animal health and Bayesian analysis, Vaccine, 281, 17717.
welfare standards, particularly for a disease
which has resulted in high economic losses Project website
and poses potential risks to human health. http://www.flu-test.eu/
A key impact of FLUTEST is the develop- Coordinator

ment and provision of validated diagnostic Dr Jill Banks
tests tailored to complement surveillance Virology Department
and outbreak management programmes. Animal Health and Veterinary Laboratories
The results have indicated that some Agency
technologies are not currently suited to AI Woodham Lane
detection and cannot currently be recom- Addlestone, Surrey, KT15 3NB
mended, and the project has provided con- UNITED KINGDOM
fidence that current surveillance protocols jill.banks@ahvla.gsi.gov.uk
are largely fit for purpose.
Comin, A., Klinkenberg, D., Marangon, S., Tof-
fan, A., Stegeman, A., Transmission dynam-
ics of low pathogenicity avian influenza
infections in turkey flocks, (submitted).
Gonzales, J.L., Elbers, A.R.W., Bouma, A.,

Koch, G., De Wit, J.J., Stegeman, J.A. (2010),
Partners
Central Science Laboratory Dr Neil Boonham n.boonham@csl.gov.uk
(FERA), Sand Hutton,
York YO41 1LZ
UNITED KINGDOM
Laboratorio di Virologia, Dr Stefano Marangon smaragon@izsvenezie.it
Istituto Zooprofilattico Speri-
mentale delle Venezie, Viale
dellUniversit, 10, 35020
Legnaro (PD)
ITALY
Central Institute of Dr Guus Koch guus.koch@wur.nl
Wageningen (UR-CVI),
PO Box 2004, Houtribweg
39, Lelystad 8203 AA
NETHERLANDS
Onderstepoort Veterinary Dr Celia Abolnik AbolnikC@arc.agric.za
Institute
SOUTH AFRICA
FLI Insel Riems, Boddenblick Dr Martin Beer martin.beer@fli.bund.de
5a, 17493 Greifswald-Insel Head of the Institute of
Riems Diagnostic Virology
GERMANY
Departments of Virology, Prof. Sndor Belk sandor.belak@sva.se
Ulls vg 2B DVM, PhD, DSc, Head of
SE-751 89 Uppsala the Joint R & D Division
SWEDEN
Cambridge University Prof. Piero Migliorato, pm@eng.cam.ac.uk
Engineering Department Professor of Physical
University of Cambridge Electronics,
UNITED KINGDOM Polysilicon TFT Group
Sanitaire des Aliments,
AFSSA-site de Ploufragan,
B.P. 53, Ploufragan 22440
FRANCE
National Veterinary Dr Poul Henrik Jr- pojo@vet.dtu.dk
Institute gensen,
DENMARK Head of Avian Virology
section
Institute of Virology and Dr Martin Hofmann Martin.Hofmann@ivi.admin.ch
Immunoprophylaxis, Sense-
mattstrasse 293,
3147-Mittelhusern
SWITZERLAND

University of Utrecht, Fac- Dr J Arjan Stegman j.a.stegeman@vet.uu.nl
ulty of Veterinary Medicine,
Androclus building, Yalelaan
1, De Uithof, 3584 CL Utrecht.
Postal address: Faculty bureau
of Veterinary Medicine
P.O. Box 80.163
3508 TD Utrecht
NETHERLANDS
Cepheid Europe, Vira-Solelh, Dr Niklas Finnstrom nfinnstrom@cepheideurope.fr
Maurens-Scopont FRANCE
Kantonales Laboratorium Claudia Bagutti Claudia.Bagutti@bs.ch
Basel-Stadt
SWITZERLAND
Forsite Diagnostics Ltd. Sand Dr Chris Danks c.danks@forsitediagnostics.com
Hutton
York YO41 1LZ
UNITED KINGDOM
Leeds Innovation Centre 103 Dr Paul Ko-Ferrigno P.KoFerrigno@leeds.ac.uk
Clarendon Road
Leeds LS2 9DF,
UNITED KINGDOM
Bath University, Department Dr Pedro Estrela P.Estrela@bath.ac.uk
of Electronic & Electrical
Engineering, University of Bath,
Bath BA2 7AY
UNITED KINGDOM
[FLU-LAB-NET]
Development and
enhancement of laboratory
networks for avian influenza
Summary laboratory capacity and capabilities for
Acronym:
FLU-L AB-NET provides new opportuni- AI diagnosis and surveillance. The rapidly FLU-LAB-NET
ties for enhancement and reinforcement of expanding growth in AI work has high-
Project number:
the Community Reference Laboratory and lighted the benefits of closer collabora-
044453
National Reference Laboratory network for tion within existing AI laboratory networks.
avian influenza (AI) within the European Spread to humans has also highlighted real EC contribution:
EUR 930 000
Union. This contributes to the strengthening, concerns of the pandemic potential of H5N1
harmonisation and development of labora- and, more recently, the emergence and Duration:
tory and diagnostic methods, coordination global spread of the pandemic H1N1(2009) 48 months
of research efforts and sharing of expertise. virus in humans has reinforced the need Start date:
Rapid responses to national and global emer- for closer integration between veteri- 1 March 2007
gencies with data sharing have been key nary and public health laboratories and Instrument:
areas of exploitation, contributing to aEuro- demonstrated the benefits in real time. Coordination action
pean laboratory task force capability for AI
in animal species. Rapid, formal interactive
communications are addressed through web- Aim
based forums. Laboratories involved in influ- To share and exchange methodological, viro-
enza research in domestic mammals also logical, genetic, epidemiological and clinical
participate. FLU-LAB-NET has fostered for- information on influenza. The network will
mal links with corresponding human, swine present up-to-date, quality information on
and equine influenza networks. FLU-LAB-NET influenza activities for scientists, policymak-
provides opportunities for identification and ers, professionals and the public. It will also
development of the complementarities of encourage the identification of duplicate
global, multi-disciplinary influenza research areas of work including surveillance and
programmes. Strategically important third- research projects at a European level.
country and INCO partners continue to be
included in this network, in order to raise lab-
oratory standards and benefit from knowl- Expected results
edge sharing. This will promote greater trust, Enhancement and reinforcement of the
understanding and early access to informa- existing Community Reference Labora-
tion that may be of importance to both vet- tory and National Reference Laboratory
erinary and public health in the EU. network for avian influenza (AI) within
the European Union Member States (EU
MS) achieved and ongoing.
Problem Development and implementation of
The global H5N1 HPAI crisis has resulted web-based, global interactive communi-
in a significantly increased demand for ties facilitating rapid, formal interactive
communications forums achieved and sharing, are key areas of exploitation, con-
ongoing. tributing to a European laboratory task
Strengthening harmonisation and devel- force capability for AI in animal species.
opment of laboratory and diagnos- FLU-LAB-NET provides opportunities for the
tic methods, coordination of research identification and development of the com-
efforts, and sharing of expertise plementarities of both EU and global multi-
achieved and ongoing. disciplinary influenza research programmes.
Facility for rapid responses to national This promotes greater trust, understanding
and global emergencies, with data and early access to information that may be
sharing achieved and ongoing. of importance to both veterinary and public
Extension of knowledge sharing and health in the EU.
laboratory support to strategically
impor tant third-countr y and INCO
partner laboratories achieved and References/publications
ongoing. Anon (2007), Development and Enhance-
Participation of laboratories involved ment of Laboratory Networks For Avian
in influenza research in domestic Influenza FLU-L AB-NET, Influenza
mammals achieved and ongoing. Research EU-Funded Projects 200107,
Fostering of formal links and coordin Luxembourg, Office for Official Publi-
ation with corresponding human, swine cations of the European Communities,
and equine influenza networks 2007 (January), 96 pp., ISBN 978-92-79-
achieved and ongoing. 05420-4, pp. 7475 (http://ec.europa.eu/
research/health/poverty-diseases/doc/
influenza-research_en.pdf).
By reinforcing and enhancing the exist- Anon (2008), FLU-L AB-NET: Enhanc-
ing Community Reference Laboratory and ing Global Avian Influenza Networks, VLA
National Reference Laboratories network Annual Review 2007/8.
for AI within the EU, FLU-LAB-NET con
tinues to facilitate improvements and har- Brown, I.H. (2008), FLU-LAB-NET Pro-
monisation of laboratory and diagnostic gress Report, Joint 14th Annual Meeting of
methods. Following the development and the National Laboratories for Avian Influ-
implementation of FLU-LAB-NET as a web- enza and Newcastle Disease of European
based, global interactive community, the Union Member States, Brussels, Belgium,
EU Member State, third-country and INCO 911 April 2008 (http://ec.europa.eu/food/
partner laboratories have facilities to opti- animal/diseases/controlmeasures/avian/
mise rapid, formal interactive communica- docs/8_FLU-LAB-NETCRL14-final.pdf).
tions forums. In turn, the network continues
to be extended to participating laboratories Brown, I.H., Banks, J. (2009), Influenza
involved in influenza research in domes- activities at VLA-Weybridge relevant to the
tic mammals, and provide a hub fostering human-animal interface (inc. FLU-LAB-
formal links and coordination between cor- NET), CMO-CVO joint meeting on Influenza,
responding human, swine and equine influ- Brussels, 30 October 2009.
enza networks.
Project website
In addition, FLU-LAB-NET allows for the http://www.flu-lab-net.eu/
coordination of research efforts and data
exchange, as well as development and Keywords
sharing of expertise. Rapid responses to avian influenza, laboratory, network,
national and global emergencies, with data FLU-LAB-NET
Coordinator
Prof. Ian H. Brown
Veterinary Laboratories Agency
Weybridge, Woodham Lane
New Haw, Addlestone
Surrey
KT15 3NB
UNITED KINGDOM
i.h.brown@vla.defra.gsi.gov.uk
Partners
Veterinary & Agrochemical Dr Thierry van den Berg thvan@var.fgov.be
Research Centre Department
of Small Stock Diseases, Avian
Virology & Immunology Unit,
99 Groeselenberg, 1180 Brussels
BELGIUM
Danish Institute for Food and Poul Henrik Jrgensen phj@dfvf.dk
Veterinary Research, Hangoevej 2
Aarhus N,
DENMARK
Friedrich-Loeffler-Institute, Dr Martin Beer martin.beer@fli.bund.de
Federal Research Institute for
Animal Health (FLI) Insel Riems,
Boddenblick 5a, 17493 Greifswald-
Insel Riems,
GERMANY
Estonian Veterinary and Food Mr Ants Jauram ants.jauram@vetlab.ee
Laboratory, Kreutzwaldi 30, Tartu,
ESTONIA
Ministry of Rural Development and Vasiliki Rousi vrousi@yahoo.gr
Food, Centre of Athens Veterinary
Institutions, 25 Neapoleos Street
Athens,
GREECE
Sanitaire des Aliments,
AFSSA-site de Ploufragan, B.P. 53,
Ploufragan 22440,
FRANCE
Central Veterinary Research Pat Raleigh pat.raleigh@agriculture.gov.ie
Laboratory, Department of
Agriculture and Food Laboratories,
Backweston Campus, Staccumny
Lane, Celbridge, Co. Kildare
IRELAND

Istituto Zooprofilattico Ilaria Capua icapua@izs.venezie.it
Sperimentale delle Venezie,
Laboratorio di Virologia, Viale
dellUniversit, 10, 35020 Legnaro
(PD),
ITALY
Veterinary Services, 1417 Dr Kyriacos Georgiou director@vs.moa.gov.cy
Athalassa, Nicosia, CYPRUS
National Diagnostic Centre of Food Mrs Ieva Rodze ieva.rodze@ndc.gov.lv
and Veterinary Service, Lejupes
str. 3, Riga,
LATVIA
Central Veterinary Institute, Prof. Vilmos Plfi palfiv@oai.hu
Tbornok utca 2.,
Budapest,
1149,
HUNGARY
Central Institute for Animal Disease Dr Ben Peeters ben.peeters@wur.nl
Control, Division of Virology,
PO Box 2004, Houtribweg 39,
Lelystad 8203 AA,
NETHERLANDS
Dr Eveline Wodak, AGES IVET Dr Eveline Wodak eveline.wodak@ages.at
Moedling, Robert Koch Gasse 17,
Moedling,
AUSTRIA
National Veterinary Research Dr Zenon Minta zminta@piwet.pulawy.pl
Institute, Department of Poultry
Diseases, Al Partyzantw 57,
24-100 Pulawy,
POLAND
Laboratorio Nacional de Dr Miguel A.S.P. miguel.fevereiro@lniv.
Investigacao Veterinarial, Estrada Torres Fevereiro min-agricultura.pt
de Benfica 701, Lisboa,
PORTUGAL
University of Ljubljana Veterinary Dr Olga Zorman Rojs olga.zorman-rojs@vf.uni-lj.si
Faculty, Institute of Poultry Health
and Protection, Gerbieva 60,
1000 Ljubljana,
SLOVENIA
State Veterinary Institute Zvolen, Miroslav Moji mojzis@svuzu.sk
National Reference Laboratory
for AI and ND, Poddrahami 918,
960 86 Zvolen,
SLOVAKIA
Finnish Food Safety Authority Evira, Liisa Sihvonen liisa.sihvonen@evira.fi
Virology Unit, Mustialankatu 3,
FI-00790 Helsinki,
FINLAND
The National Veterinary Institute, Prof. Sndor Belk sandor.belak@bvf.slu.se
Ulls vag 2B, Uppsala,
SWEDEN

Agri-food and Biosciences Institute, David Graham david.graham@dardni.gov.uk
Veterinary Sciences Division,
Stoney Road, Stormont, Belfast
BT4 3SD,
NORTHERN IRELAND
National Veterinary Institute, Christine Monceyron christine.monceyron-jonassen@
Section for Virology and Serology, Jonassen vetinst.no
PO Box 8156 Dep., 0033 Oslo
NORWAY
National Diagnostic Research Dr Gabriela gvgoujgoulova@abv.bg
Veterinary Medical Institute, 15 Goujgoulova
Pencho Slaveikov Blvd, Sofia 1606,
NRL of AI and NDV, 190 Lomsko
shose Blvd., 1231 Sofia,
BULGARIA
Institute for Diagnosis and Animal Dr Aurelia Ionescu ionescu.aureila@idah.ro
Health, Bucharest, ROMANIA aureliaonescu@yahoo.com
National Reference Centre for Prof. Richard Hoop rhoop@vetbakt.unizh.ch
Poultry Diseases, Dept of Poultry
Diseases, Institute of Veterinary
Bacteriology, Winterthurerstr. 270,
8057 Zrich,
SWITZERLAND
Investigational and Diagnostic Dr Wlodek Stanislawek wlodek.stanislawek@maf.govt.nz
Centre-Wallaceville, Biosecurity
New Zealand, Ministry of Agriculture
and Forestry, PO Box 40742,
5018 Upper Hutt,
NEW ZEALAND
Kenya Agricultural Research Dr Yatinder Singh karibiotech@kari.ng;
Institute, Biotechnology Center, P.O. Binepal ybinepal@yahoo.com
Box 57811-00200, Nairobi, KENYA
Veterinary Faculty Poultry Emina Reidbegovi eminar@vfs.unsa.ba
Centre, Zmaja od Bosne
90, 71000 Sarajevo
BOSNIA AND HERZEGOVINA
Clinics Laboratory, School of Prof. Chukwudozie cezeokoli@fms.uwi.tt
Veterinary Medicine, Faculty of Daniel Ezeokoli ezeokoli01@yahoo.com
Medical Sciences, The University
of the West Indies, St. Augustine,
Trinidad and Tobago,
WEST INDIES
Animal Influenza Laboratory of Prof. Hualan Chen hlchen1@yahoo.com
Ministry of Agriculture, Harbin
Veterinary Research Institute,
Chinese Academy of Agricultural
Sciences, 427 Maduan Street,
Harbin 150001,
CHINA

National Reference Laboratory for Dr Khalid Naeem NAEEM22@isb.comsats.net.pk
Poultry Diseases, National Agricul-
tural Research Centre, Park Road,
Islamabad 45500,
PAKISTAN
Croatian Veterinary Institute, Vladimir Savi vsavichr@yahoo.com
Poultry Centre, Heinzelova 55,
10000 Zagreb,
CROATIA
Animal Health Trust, Lanwades Debra Elton debra.elton@aht.org.uk
Park, Kentford, Newmarket, Suffolk,
CB8 7UU, UNITED KINGDOM
University of Gent, Sint-Pieters Kristen Van Reeth kristien.vanreeth@UGent.be
nieuwstraat 25, Gent,
BELGIUM
Laboratorio Central de Veterinaria Concepcin cgometze@mapya.es
(Algete), Ctra. De Algete, km. 8, Gmez-Tejedor
28110 Algete (Madrid),
SPAIN
Federal Centre for Animal Health, Dr Andriyasov Artem andriyasov_av@arriah.ru
ARRIAH, Yurevets, Vladimir,
600901,
RUSSIAN FEDERATION
Bornova Veterinary Control and Fethiye Coven covenfethiye@hotmail.com
Research Institute, Osman Kibar
Meydani, Ege. Univ., Kavsagi,
Bornova/Izmir, 350010,
TURKEY
Centre de Recherche public Claude Muller claude.muller@ins.etat.lu
de la Sant
84, rue Val Fleuri
1526 Luxembourg
LUXEMBOURG
Ministry of Rural development George Georgiades gkgeorgi@otenet.gr
and Food, Centre of Thessaloniki
Veterinary Institutes,
26th October St, 80,
Thessaloniki 546 27,
GREECE
[FLUTRAIN]
Training and technology transfer

of avian influenza diagnostics
and disease management skills
Summary both for serological and virological
The ongoing worldwide avian influenza (AI) diagnosis that were developed in three Acronym:
FLUTRAIN
crisis has highlighted the need for com- European projects namely AVIFLU, LAB-
prehensive training and the transfer of ON-SITE and FLUAID. In 2009, the global Project number:
technology to accession and International outbreaks of pandemic H1N1 highlighted 044212
Cooperation Countries (INCO) with the clear the unpreparedness of many laborato- EC contribution:
goal of aiding these countries in combat- ries to deal with such a crisis particularly EUR 1 809 133
ing AI with the provision of the most up-to- if the virus infected pig populations. In Duration:
date diagnostic and disease management order to address this issue, the project 51 months
procedures. The FLUTRAIN project aimed to was amended so that important data
Start date:
fulfil this requirement at two levels. on the immunity of pig populations to 1 March 2007
pandemic H1N1 could be generated and
Instrument:
It resolved the need for training by pro- transferred to laboratories in developing Coordination action
viding two workshops over the duration countries.
of the project that allowed experts in the
AI field to pass on their expertise in the The consortium was made up of 11 mem-
diagnosis and management of AI to par- bers (including two SMEs) in addition to
ticipants from accession and INCO coun- eight associated partners, many of whom
tries. Follow-on training opportunities were recipients of training.
were provided in partner laboratories
in order to consolidate the information
and practical experience gained during Problem
the workshops. In addition, FLUTRAIN Avian influenza, and more recently pandemic
identified and supplied funds for one-off H1N1, infections have increased in rele-
specific support missions that targeted vance both from animal and human health
specific AI problems in recipient coun- perspectives. With the extension of the
tries. The project partners have also H5N1 epidemic from Asia to eastern Europe
developed an e-learning course (E-Flu) and Africa and the global spread of H1N1,
for AI that was designed to support vet- notwithstanding the efforts of international
erinary administrations in training newly organisations, there is clear evidence that in
recruited staff involved in AI prevention, the current situation, attempts to stop the
diagnosis and control activities. infections progress are insufficient. This
The transfer of technology to accession has highlighted the need for comprehensive
and INCO countries via the provision of training and the transfer of technology to
new, simplified and cost-effective diag- accession and INCO countries with the clear
nostic methods and reagents. It also goal of aiding these countries in combating
involved the transfer of deliverables, AI, and possibly H1N1 in swine populations,
using the most up-to-date diagnostic and 7. Generation of online course on avian influ-
disease-management procedures. enza management and control (E-Flu).
8. Generation of important tools and
information on the immunity of pig
Aim populations to pandemic H1N1.
Avian influenza infections caused by several 9. Generation of important tools and infor-
subtypes are endemic in vast areas of the mation on avian H1, H2 and H3 subtype
world, particularly in developing countries viruses
and countries where poverty is widespread.
Under these circumstances, AI infections Project website
are very difficult to control, due to the lack http://www.flutrain.eu
of funds to train staff, produce or purchase
diagnostic reagents or apply modern diagnos- Publications
tic technologies. The objective of FLUTRAIN Capua, I., Kajaste-Rudnitski, A., Bertoli, E.,
was therefore to bridge the gap of knowledge Vicenzi, E., Pandemic vaccine prepared-
between EU scientists and colleagues in AI- ness have we left something behind?,
affected countries. This project also had the PLoS Pathogens, June 2009, 5(6): e1000482.
objective of supporting countries affected by
AI infections through training and the trans- Fusaro, A., Joannis, T., Monne, I., Salviato, A.,
fer of knowledge and technology. Yakubu, B., Meseko, C., Oladokun, T., Fassina,
S., Capua, I., Cattoli, G., Introduction into
As a result of the global H1N1 (2009) Nigeria of a distinct genotype of avian influ-
pandemic, FLUTRAIN also covered impor- enza virus (H5N1), Emerging Infectious Dis-
tant aspects of swine influenza. In 2009, eases, March 2009, 15(3): 4457.
it was not known to what extent pigs were
immune to infection with the pandemic Monne, I., Joannis, T.M., Fusaro, A., De Ben-
H1N1 virus. Indeed, many developing coun- edictis, P., Lombin, L.H., Ularamu, H., Egbuji,
tries do not have the expertise or facilities A., Solomon, P., Obi, T.U., Cattoli, G., Capua, I.,
to answer such questions and so, using the Reassortant avian influenza virus (H5N1) in
European situation as a model, data were poultry, Nigeria, 2007, Emerging Infectious
generated to determine the cross reactiv- Diseases, April 2008, 14(4): 63740.
ity of the EU swine population to pandemic
H1N1 (2009). Monne, I., Ormelli, S., Salviato, A., De Bat-
tisti, C., Bettini, F., Salomoni, A., Drago, A.,
Zecchin, B., Capua, I., Cattoli, G., Develop-
Results ment and validation of a one-step real-
1. Data on alternative methods for virus time PCR assay for simultaneous detection
isolation and detection. of subtype H5, H7, and H9 avian influenza
2. Development of alternative methods for viruses, Journal of Clinical Microbiology,
reagent production and improvement. May 2008, 46(5): 176973.
3. Training of diagnosticians and scientists
in EU laboratories. Pizzuto, M., De Benedictis, P., Maniero, S.,
4. Inter national training wor k shops Toson, M., Dundon, W.G., Seck, B., Capua,
addressing general and specific topics I. (2011), Improving heat stability of
related to AI. haemagglutinating antigens for avian
5. Two international meetings in collabora- influenza, Biologicals (in press).
tion with OIE, FAO and WHO to discuss AI
at the human-animal interface. Keywords
6. Ad hoc support and training missions in avian influenza, training, technology
Egypt, Senegal and Sierra Leone. transfer, INCO
Coordinator Legnaro 71
Dr Ilaria Capua DVM, PhD ITALY
OIE\FAO and National Reference Laboratory icapua@izsvenezie.it
for Avian Influenza and Newcastle Disease
Istituto Zooprofilattico Sperimentale delle
Venezie
Partners
Veterinary Laboratories Agency, Virol- Dr Jill Banks j.banks@vla.defra.gsi.gov.
ogy Dept, Woodham Lane, Addlestone, uk
Surrey,
UNITED KINGDOM
Central Veterinary Institue (CVI), Dr Guus Koch Guus.Koch@wur.nl
Houtribweg 39 8221 RA Lelystad THE
NETHERLANDS
Veterinary and Agrochemical Research Dr Thierry van den Berg Thierry.vandenBerg@var.
Center Brussels, fgov.be
BELGIUM
Ghent University, Faculty of Veterinary Dr Kristien van Reeth kristien.vanreeth@UGent.be
Medicine, Merelbeke,
BELGIUM
Swedish University of Agricultural Sci- Dr Sandor Belak sandor.belak@bvf.slu.se
ences
SWEDEN
Istituto Zooprofilattico Sperimen- Dr Maura Ferrari maura.ferrari@bs.izs.it
tale della Lombardia e dellEmilia-
Romagna, Brescia,
ITALY
Technical University of Denmark, Anker Dr Poul Henrik PHJ@vet.dtu.dk
Engelundsvej 1, bygning 101 A, Lyngby,
DENMARK
Dept of Public Health, University of Dr Alessandra Piccirillo alessandra.piccirillo@
Padua, unipd.it
ITALY
Capture Productions Ltd, Carlow Mr Kevin McDermott kevin.mcdermott@capture.
IRELAND ie
Svanova Biotech AB, Uppsala SWEDEN Dr Malik Merza malik.merza@svanova.com
Department of Virology, Division of Dr Wendy Barclay w.barclay@imperial.ac.uk
Investigative Science, Faculty of Medi-
cine, Wright Fleming Institute, Norfolk
Place, London, W2 1PG,
UNITED KINGDOM
Associated Partners
US Department of Agriculture , South- Dr Dennis Senne dennis.a.senne@aphis.
east Poultry Research Laboratory, Iowa, usda.gov
UNITED STATES

Institute for Diagnosis and Animal Dr Ionescu Aurelia daureliaionescu@yahoo.
Health Bucharest, com
ROMANIA
Croatian Veterinary Institute Zagreb, Dr Vladimir Savic vsavichr@yahoo.com
CROATIA
University of Ljubljana, Veterinary Dr Olga Zorman-Rojs olga.zorman-rojs@vf.uni-lj.
Faculty, Ljubljana, si
SLOVENIA
Firat University, Veterinary Medicine, Dr Aykut Ozdarendeli aozdarendeli@firat.edu.tr
TURKEY
National Research Veterinary Institute Dr Tony Joannis tmjoannis@yahoo.com
Vom,
NIGERIA
National Center for Veterinary Diag- Dr Thanh Long To thanhto@fpt.vn
nosis , Dept of Animal Health 1178th
Lane , Giai-Phong St, Dong-da Hanoi,
VIETNAM
National Diagnostic and Research Dr Georgi Georgiev georgivet@yahoo.com
Veterinary Medical Institute BULGARIA
[ConFluTech]
Capacity-building for the control

of avian influenza through
technology transfer and training
Acronym: Summary Besides the fact that a number of countries
ConFluTech Avian influenza (AI) or bird flu is a highly were surprised by the outbreaks, an even
contagious viral infection which can affect greater number of developing countries do
Project number:
44462 all species of birds and can manifest itself not have adequate tools to detect and dif-
in different ways depending mainly on the ferentiate HPAI and are lacking experience
EC contribution:
pathogenicity of the virus involved and on to manage the outbreak of the disease.
EUR 547 255
the species affected. The highly patho- Thus, there is an urgent need for technol-
Duration: genic avian influenza (HPAI) virus causes ogy transfer and training. To fill these gaps,
60 month
serious disease with high mortality (up to the partners of this consortium will:
Start date: 100%) notifiable to the OIE. Worry-
1 July 2007 ingly, HPAI has been shown to infect and (a) organise technical workshops to facili-
Instrument: cause death in humans. Up to now, a total tate technology transfer particularly in
specific support of 103 deaths have been recorded due to the field of molecular diagnostic tools
action HPAI infection in a number of countries for pathogen detection and differenti
such as Vietnam, Turkey and Iraq (WHO, ation, to reinforce epidemiological
21 March 2006). analysis for monitoring and modelling
of avian influenza especially and to (b) Standardisation and validation of diag-

respond to outbreaks of infectious dis- nostic tools according to OIE instruction;
eases of livestock in general; (c) Epidemiological tools for monitoring and
(b) provide training through organisation modelling avian influenza outbreaks;
of seminars and short-term courses in (d) Management of disease outbreaks.
well-qualified laboratories of a number
of EU Member States;
(c) organise technical workshops, courses Potential applications
and training in the INCO target countries The following technical and scientific
to improve the technical experimental advances are expected:
level of the staff and laboratories in
charge of livestock infectious diseases. (a) transfer of knowledge and technology
for surveillance systems to exploit and
understand disease transmission path-
Problem ways, establishment of reliable indica-
Avian influenza is a highly contagious viral tors and tools for disease-monitoring
infection which can affect all species of activities (analytical and modelling) to
birds and can manifest itself in different describe the present situation and pre-
ways depending mainly on the pathogen dict the future course of AI as well as for
icity of the virus involved and on the spe- a better response to diseases;
cies affected. The highly pathogenic avian (b) standardised diagnostic tools and
influenza (HPAI) virus causes serious dis- methods;
ease with high mortality (up to 100 %). (c) tools in decision-making, communication
Worryingly, HPAI has been shown to infect and information systems.
and cause death in humans in a number of
countries such as Iraq, Turkey and Vietnam.
Besides the fact that a number of countries Activities
were surprised by the outbreaks, an even A number of activities, including technical
greater number of developing countries do workshops, courses, training and collabor
not have adequate tools to detect and dif- ations were organised at both national and
ferentiate HPAI and are lacking experience regional levels.
to manage the outbreak of the disease.
Thus there is an urgent need for technology In the context of collaboration, links were
transfer and training. established to:
authorities in charge of veterinary and

Aim human public health, research centres
The overall aim of this project is to facili- and universities in many countries;
tate technology transfer and training to a number of other EU-funded pro-
promote capacity-building in INCO (Interna- jects such as FLUTRAIN, LAB-ON-SITE,
tional Cooperation) target countries, with a Income, ASEM-Dialog, EPIZONE, etc.,
particular emphasis on countries that border dealing with avian influenza or other
the EU, for better control of avian influenza infectious diseases;
and general outbreaks of infectious diseases FAO, European Society for Veterinary
in livestock. This will be achieved through Virology, CIRAD and the FGI-ARIAH in the
the organisation of technical workshops and Russian Federation.
training courses in the following fields:
Through these links, it was possible to
(a) Molecular diagnostic tools for pathogen extend the activities of ConFluTech to cen-
detection and differentiation; tral and eastern Europe, the Middle East,
Central Asia and North Africa. Thus, sem and activities as well as six standard
inars and technical workshops for veterinary operation procedures (SOP).
staff from Croatia, Czech Republic, Estonia, D is t r i b u t i o n of C D s c o n t a i n i n g
Hungary, Lebanon, Lithuania, Macedonia, presentations and protocols of the
Mauritania, Moldova, the Palestinian workshops.
National Authority, Serbia, Slovakia, Slovenia Distribution of a special issue of Trans-
were successfully organised. The seminars boundary and Emerging Diseases,
and technical workshops held by ConFluTech Vol.55, number 56, August 2008, con-
covered areas of great relevance for the taining the proceedings of the Income
control of the diseases including: project held at the Lisbon meeting,
2007; supplemental issue of Vaccine,
biosecurity at farm and diagnostic Vol. 26, Suppl. 6, December 2008 con-
laboratory level; taining proceedings of the ICTTD-3
e p i d e m i o l o g y, m o n i t o r i n g a n d meeting held in Borstel, 2007.
surveillance tools; Maintenance of a specific website for
sampling and transport of biological this purpose (http://www.conflutech.net).
material;
preparation of awareness booklets in
different languages. Keywords
avian influenza, polymerase chain reaction,
To cover these fields, experts in the fields epidemiology, disease outbreak
of management of poultry and poultry dis-
eases, laboratory diagnostics, biosecurity,
outbreak management, epidemiology and Coordinator
disease monitoring and surveillance were Prof. Jabbar Ahmed
invited. These included Prof. Hafez Ahmed Research Center Borstel
Hafez, Head of the Institute of Poultry Parkallee 22
Diseases from the Free University of Ber- 23845 Borstel GERMANY
lin and President of the World Society for Tel. +49 4537188428
Poultry, Dr Abdulwahab from the same E-mail: jahmed@fz-borstel.de
institute, Dr Filip Claes, Institute of Tropical
Medicine, Antwerp and Dr Mohammed El-
Nator, Veterinary Faculty, Jordan Technical
University, Irbid.
In summary, a total of 22 technical

workshops and workshops were held on a
national or regional basis in the following
countries: Armenia, Austria, Bulgaria, Geor-
gia, Germany, Greece, Iraq, Jordan, Mauri-
tania, Romania, Russia, Slovakia, Sweden,
Syria and Turkey.
Dissemination activities
The following information has been
disseminated.
Distribution of a booklet, containing

information about the project aims
Partners
Agricultural Sciences, Ulls vg 2B Prof. Sndor Belk sandor.belak@sva.se
SE-751 89 Uppsala
SWEDEN
Catedrtico de Sanidad Animal, Prof. J.M. jmvizcaino@vet.ucm.es
Universidad Complutense, Dpto Snchez-Vizcano
Sanidad Animal, Avda Puerta de
Hierro s/n, 28040 Madrid
SPAIN
Cantacuzino Institute, Dr Viorel roinfluenza@cantacuzino.ro
ROMANIA Alexandrescu
Pendik Veterinary Control, Cad Dr Iyisan, Ayse selmai@superonline.com
No 10, 81480 Pendik, Istanbul Selma
TURKEY
State Veterinary Inspection Dr Hokobyan horhannes_Hokobyan@yahoo.com
ARMENIA Hohannes
Ganja State Agricultural Academy, Prof. Jafarov sabinashukurova@yahoo.com
AZERBAIJAN Mamedtagi
Georgian State Agricultural Prof. Levan
University, Tbilisi, Makaradze
GEORGIA
University of Tehran, Faculty Ass. Prof. Parviz pshayan@ut.ac.ir
of Veterinary Medicine, Shayan
Dept of Parasitology,
IRAN
Dohuk University and Research Dr Lokman Taib lokman_ommer@yahoo.com
Center, Faculty of Veterinary Omer
Medicine, Dohuk,
IRAQ
Faculty of Veterinary Medicine, Prof. Dr Darem spana@net.sy
Al-Baath University, Hama Tabbaa
SYRIA
Faculty of Agriculture and Prof. Dr Labib Sharif sharif@just.edu.jo
Veterinary Medicine, The Jordan
University of Science and
Technology,
JORDAN
Production Agricultural, Prof. Rogdakis erog@aua.gr
University of Athens, Laboratory Emmanouil
of General and Special Animal
Technology
GREECE
National Reference Laboratory of Ass. Prof. Dr Rositsa kotseva@ncipd.netbg.com
Influenza and Acute Respiratory Kotseva
Diseases, 26 Yanko Sakazov Blvd,
Sofia,
BULGARIA
Animal Production and Health Dr Adama Diallo a.diallo@iaea.org
Section, FAO/IAEA Joint Division,
Wagramer Strasse 5, P.O. Box 100,
1400 Vienna,
AUSTRIA
4.2. Swine flu

[ESNIP 2]
European Surveillance
Network for Influenza in Pigs 2
Summary to ensure broad protection. So far, there
Acronym: The European Surveillance Network for had been little surveillance for SIVs and
ESNIP 2
Influenza in Pigs (ESNIP) 2 maintained and there was a lack of standardised reagents
Project number: expanded a surveillance network that was and protocols for subtyping and antigenic/
022749 established during a previous EC concerted genetic characterisation of SIV. A rapid and
EC contribution: action (ESNIP 1, QLK2-CT-2000-01636). The simple test would be valuable. Finally, pigs
EUR 300 000 main aim was to gain a better understand- are known to be susceptible to infection
Duration: ing of the epidemiology and evolution of with both human and AI viruses, but the
36 months swine influenza virus (SIV) in Europe through true public health risk of this was unknown,
Start date:
an organised surveillance programme and due to a lack of screening of pigs for influ-
1 January 2006 extensive antigenic/genetic characterisation. enza virus from other hosts.
The data were used to improve the diagno-
Instrument:
Coordination action sis of SIV by updating the reagents used in
classical techniques and by the development Aim
of a rapid molecular test. The virus bank The first objective of ESNIP 2 was to expand
and electronic database of ESNIP 1 were our knowledge of the epidemiology and evo-
expanded. In addition, we performed a sero- lution of SIVs in Europe, and to apply this
logical monitoring of swine for avian influ- knowledge to optimise diagnostic techniques
enza (AI) viruses. Influenza viruses currently for swine influenza. Therefore, we aimed to:
circulating in European swine were compared
with those in avian species and humans as keep track of major changes in the
well as with influenza viruses circulating in epidemiology of SIV in Europe;
southern China and the United States. study the extent of antigenic and genetic
evolution of SIVs;
improve the diagnosis of SIV;
Problem expand the SIV bank and electronic
The epidemiology of swine influenza in database of ESNIP 1.
Europe has become particularly complex. At
least three SIV subtypes (H1N1, H3N2 and The second objective of ESNIP 2 was to pro-
H1N2) are circulating and new reassortants vide insights into the public health risk of
between these subtypes have been detected influenza in swine by monitoring swine for
occasionally. The heterogeneity in SIVs has AI viruses and by comparison of influenza
important implications for diagnosis and viruses in swine and in human populations.
control. The strains used in serodiagnostic Therefore, we aimed to:
tests need to be matched to the current
epidemic viruses, and inactivated SIV vac- screen European swine populations for
cines should contain all prevailing subtypes the circulation of AI viruses;
compare the influenza situation in swine vaccination strategies on individual swine

with that in humans and birds, and farms. In addition, it provided insight into
ensure dissemination of information to the SIV subtypes that are currently circu-
human and AI researchers. lating in Europe and their antigenic charac-
teristics in comparison with vaccine strains.
Major outcomes are: These insights are crucial in the decision
whether changes in the vaccine strain com-
maintenance and expansion of the pig position may be required. Furthermore,
surveillance network; ESNIP 2 guaranteed transparency of ani-
data about the prevalence and circula- mal disease status worldwide. It helped to
tion of SIV subtypes in swine in Europe collect, analyse and disseminate veterinary
and comparison with the situation in scientific information to Member States in
China and the United States; order to improve their methods for control
antigenic and genetic characterisation of of influenza.
the isolated SIVs;
recommendations for the reagents to be
used in classical diagnostic tests for SIV; Project website
standardisation and validation of http://www.esnip.UGent.be
rapid tests for the diagnosis and
characterisation of SIVs; Keywords
maintenance and expansion of the SI influenza, swine, surveillance, evolution,
virus bank and electronic database; diagnosis, public health risk
first approaches in the development of
aserologic assay for AIVs in swine; Coordinator
formal interaction between the swine, Prof. Kristien Van Reeth
avian and human surveillance networks. Ghent University
Laboratory of Virology
Potential applications Salisburylaan 133
ESNIP 2 improved and facilitated the 9820 Merelbeke
diagnosis of SI and the subtyping of SIVs. BELGIUM
This is essential for a rational design of Kristien.VanReeth@UGent.be
Partners
CONTACT ORGANISATION
Dr Ian Brown Veterinary Laboratories Agency, Addlestone, UNITED KINGDOM
Dr Willie Loeffen CIDC-Lelystad, Lelystad, NETHERLANDS
Dr Emanuela Foni Istituto Zooprofilattico Sperimentale della Lombardia e
dellEmilia Romagna, Parma, ITALY
Dr Franois Madec Agence Franaise de Scurit Sanitaire des Aliments, Ploufragan,
FRANCE
Dr Mikhail Matrosovich National Institute for Medical Research, London, UNITED KINGDOM
Dr Michel Bublot Discovery Research, Merial, Lyon, FRANCE
Jaime Maldonado Veterinary Diagnostic Services DIAGNOS, Gerona, SPAIN
Prof. Ivaylo Chenchev National Diagnostic Veterinary Research Institute, Sofia, BULGARIA
Prof. Malik Peiris The University of Hong Kong, HONG KONG
Prof. Christopher W. Olsen University of Wisconsin-Madison, Madison, UNITED STATES
[ESNIP 3]
European Surveillance
Network for Influenza in Pigs 3
Summary improved pandemic preparedness and plan-
Acronym: This European Surveillance Network for ning for human influenza whilst providing
ESNIP 3
Influenza in Pigs (ESNIP) 3 will maintain an evidence base for decisions in relation
Project number: and expand surveillance networks estab- to veterinary health. The project consortium
00259949 lished during previous EC concerted actions consists of 24 participants, which contrib-
EC contribution: (ESNIP 1, QLK2-CT-2000-01636; ESNIP 2, ute a blend of different specialisms and
EUR 1 000 000 SSPE-022749). Three work packages (WP2, skills ensuring multi-disciplinary cutting-
Duration: 3, 4) aim to increase the knowledge of edge outputs. The vast majority of the part-
36 months the epidemiology and evolution of swine ners are actively working with SIV, some in
Start date:
influenza (SI) virus (SIV) in European pigs a field setting. Twenty-one participants are
1 November 2010 through organised field surveillance pro- from 11 EU Member States, seven of which
grammes (WP2). Virus strains detected were actively involved in ESNIP 2. Coop-
Instrument:
Coordination and in these programmes will be subjected to eration with partners in China and North
support action detailed characterisation both antigenically America will continue to promote a greater
(WP3) and genetically (WP4) using stand- understanding of the epidemiology of SIVs
ardised methodology. Specifically, this will at a global level.
involve timely information on genomic data
and generation of antigenic maps using
the latest technology. These analyses will Problem
provide significant and timely added value SI is enzootic in the major swine-producing
to knowledge of SIV. A strong focus will be countries of Europe and the epidemiology
monitoring spread and independent evolu- of SI in Europe is recognised to be differ-
tion of the pandemic H1N1 (2009) virus in ent from that in Asia or North America.
pigs. All these data will, in turn, be used to However, unlike human or equine viruses,
improve the diagnosis of SI by updating the organised surveillance for SIVs only began
reagents used in the recommended tech- relatively recently. Gaps in surveillance in
niques (WP2). The virus bank and electronic pigs have been criticized by public health
database that were established during officials in the light of the emergence of
ESNIPs 1 and 2 will also be expanded and the pandemic H1N1 (2009) virus putatively
formally curated with relevant SIV isolates from pigs. This large European consortium
and information for global dissemination is critical in ensuring that uncertainties over
within and outwith the consortium (WP5). the epidemiology of SI in European pigs are
ESNIP 3 represents the only organised sur- thoroughly addressed.
veillance network for influenza in pigs and
seeks to strengthen formal interactions Three antigenic and genetically distinct
with human and avian surveillance net- swine influenza virus subtypes (namely
works previously established in ESNIP 2. avian-like swine H1N1; human-like swine
A timely and transparent interaction with H3N2 (reassortant of human and avian
these networks will be a key output. These viruses); and swine H1N2 (reassortant
approaches are entirely consistent with of human and avian viruses)), have
co-circulated for many years within the 1. the standardisation of protocols for
swine population in Europe. However, the swine influenza (SI) virus (SIV) isolation,
completed ESNIP 1 and ESNIP 2 coordinat- serology, antigenic and genetic typing of
ing actions showed that the prevalence and SIV isolates;
incidence of individual subtypes may vary 2. the selection and production of refer-
from one country or region to another. For ence virus strains and (hyperimmune)
example, the H3N2 virus seems to have dis- sera these were made available to all
appeared from some regions, whereas the participants for preliminary subtyping of
H1N2 virus is becoming one of the most SIV isolates;
prevalent subtypes in others. Furthermore, 3. the establishment of a central SIV bank
new reassortant viruses, not only between with a collection of recent isolates from
the three endemic SIV subtypes, but also various geographical areas in Europe;
between SIV and seasonal human influenza 4. the establishment of an electronic data-
viruses, have occasionally been detected base with relevant information on the
during the last 10 years. Recently, isolated SIV isolates that were obtained in dif-
outbreaks of infection with the pandemic ferent countries during the life of the
H1N1 (2009) virus (pH1N1) have been network;
reported in several pig herds in the world, 5. the antigenic and genetic characterisa-
including Europe. The continued spread of tion of a number of recent H1N1, H3N2
this pandemic virus of potential swine ori- and H1N2 SIV isolates from different
gin in the human population and the dem- European countries;
onstrated high susceptibility of pigs to the 6. the organisation of a serological survey
virus makes it likely that the risk of it enter- to obtain preliminary data on the preva-
ing pig farms in Europe will increase in the lence of different SIV subtypes in various
foreseeable future. In fact, endemnicity can European countries.
be expected based on the ease of transmis-
sibility between pigs and parallels with pre-
vious human pandemic strains that became Expected results
established in global pig populations. Expan- ESNIP 3 represents the largest structured
sion and consolidation of the detection and consortium delivering coordinated surveil-
identification of swine influenza viruses in lance for influenza in pigs in Europe to date.
pig herds in Europe is necessary to provide This will be an invaluable resource to offi-
new data about potential changes in the epi- cials responsible for veterinary and public
demiology of the three endemic European health alike. The coordination action will
SIV subtypes, as well as adaptation and directly impact on the diagnosis and control
circulation of novel reassortant viruses in of SI in Europe and thus enhance the wel-
European pig herds and the introduction, and fare of swine and the profitability of swine
possible ongoing transmission, of the pH1N1 farmers. In addition, it will increase our
virus into European pigs. Recent reports of a understanding of the public health risks of
novel H3N2 human-avian reassortant virus influenza in swine. Comprehensive informa-
emerging in pigs and spreading to mink, tion relating to the epidemiology and evo-
despite no reported detection in swine, dem- lution of swine influenza in pig populations
onstrates the importance of a coordinated across Europe will be made available. This,
surveillance network for monitoring pigs in in turn, will enable a robust scientific evi-
Europe for influenza viruses. dence base to be available when assessing
public health risk from SI, directly contribut-
ing to the production of policy documents
Aim and risk assessments prepared by ECDC.
To build on the achievements of ESNIP 1 Such enhanced interaction will be timely
and 2 which were: following the emergence of the pandemic
H1N1 (2009) virus that has already been of the zoonotic potential of influenza in
detected in pigs in Europe. The continued swine. These data will be communicated to
evolution of this virus in pig populations will decision-makers and authorities in both vet-
be a key output from the project and will erinary and public health spheres. Contin-
be of strategic benefit especially for public ued evolution of the pandemic H1N1 (2009)
health. Furthermore, the authorities posi- virus, should it become established in Euro-
tion will be strengthened when handling pean pigs, will provide an evidence base to
these issues in the knowledge of a coor- policymakers on the need for enhanced dis-
dinated surveillance network addressing ease control measures for this non-notifi
influenza virus in pigs. Context at a global able disease. Policy documents have already
level will be facilitated through interaction been produced at EU level and are subject
with key global bodies (i.e. WHO) and insti- to continual review in the light of develop-
tutes furthering the EU preparedness and ments in the field. This project will directly
know-how in this subject area. inform the decision-making process through
determining the level of threat for both swine
and human health as continued evolution of
Potential applications the virus is monitored. Recommendations
Strategic impact: ESNIP 3, as the only organ- on reagents to be used for SI diagnosis and
ised surveillance network for influenza in on SIV vaccine strain composition may have
pigs, will strengthen formal interactions an impact at national or international level.
with human and avian surveillance net- Data from WP2 (serological screening of
works. Specific innovation-related outputs swine for avian influenza viruses) may have
are the improvements to and validation of a significant impact on the control of avian
rapid tests for the detection of SIVs. Sig- influenza if developments indicate changes
nificant improvements to the way SIVs are in the normal host range of notifiable avian
characterised both antigenically and genet influenza viruses. This, in turn, could have
ically will be realised through the application asignificant influence on the design of
of the latest technologies by international control strategies for avian influenza even
experts in their respective fields. This project though some provision is made in Council
will deliver the first complete antigenic maps Directive2005/94/EC.
of European SIVs using cutting-edge tech-
nologies whilst whole genome characterisa-
tion of these viruses will be carried out on References/publications
a scale not achieved previously. The latest Brookes, S.M., Irvine, R.M. et al. (2009),
tools for understanding virus evolution will Influenza A (H1N1) infection in pigs,
be used to analyse the genetic data sets. Veterinary Record, 164 (24): 760761.
Contributions to standards: Standardised Brown, I.H. (2005), Epidemiology of swine

protocols (conforming to those specified in influenza in Great Britain and emerging
the OIE Terrestrial Manual) and reference global issues, Pig Journal, 56: 145150.
reagents, for both antigenic and genetic
characterisation agreed during ESNIP 2 will Kyriakis, C.S., Brown, I. et al. (2009), Viro-
continue to be used and updated where logical surveillance and preliminary anti-
applicable. genic characterisation of influenza viruses
in pigs in five European countries from
Policy developments: Epidemiological data 2006 to 2008, Zoonoses and Public Health
on the circulation of avian/human influ- (in press).
enza viruses in swine and their compari-
son with viruses circulating in avian/human Van Reeth, K., Brown, I.H. et al. (2004),
populations will increase our understanding G e n e t ic r el at io n s hi p s , s er olo g ic a l
cross-reac tion and cross-protec tion Keywords

between H1N2 and other influenza A sub- swine inf luenza virus, sur veillance,
types endemic in European pigs, Virus serology, antigenic cartography, genetic
Research, 103: 115124. characterisation
Van Reeth, K., Brown, I.H. et al. (2008), Coordinator

Seroprevalence of H1N1, H3N2 and H1N2 Prof. Ian Brown
swine influenza viruses in seven European Animal Health and Veterinary Laboratories
countries in 200203, Influenza and other Agency
Respiratory Viruses, 2 (3): 99105. New Haw, Addlestone, Surrey
KT15 3NB
UNITED KINGDOM
Project website ian.brown@ahvla.gsi.gov.uk
http://www.esnip3.eu/
Partners and lead scientists

University of Gent, Belgium Kristien Van Reeth, Kristien.VanReeth@UGent.be
professor, Ghent
University, Faculty of
Veterinary Medicine,
Laboratory of Virology,
Salisburylaan 133, 9820
Merelbeke
BELGIUM
Agence nationale de Scurit Galle Kuntz-Simon, Unit gaelle.simon@anses.fr
Sanitaire de lalimentation, de Virologie Immunologie
lenvironnement et du travail Porcines, ANSES-LERAPP,
Zoople Les Croix, BP 53,
22440 Ploufragan
FRANCE
Istituto Zooprofilattico Speri- Emanuela Foni, Isti- emanuela.foni@bs.izs.it
mentale della Lombardia e tuto Zooprofilattico
dellEmilia Romagna Sperimentale della
Lombardia e dellEmilia
Romagna, Sezione
Diagnostica di Parma,
Via dei Mercati, 13/A,
43100 Parma,
ITALY
Danmarks Tekniske Universitet Lars Erik Larsen, DVM, lael@vet.dtu.dk
PhD, Head of Virology,
Department of Veterinary
Diagnostics and Research,
Technical University
of Denmark, National
Veterinary Institute,
Blowsvej 27, 1790
Copenhagen V

National Veterinary Research Iwona Markowska-Daniel, iwonamd@piwet.pulawy.pl
Institute Professor, Department
of Swine Diseases,
Panstwowy Instytut
Weterynaryjny PIWET,
Partyzantow 57, Pulawy,
POLAND
Ministerio de Medio Ambiente y Montserrat Agero maguero@jcasator.jazztel.es
Medio Rural y Marino Garcia, Directora Adjunta,
Laboratorio Central Vet-
erinario-Sanidad Animal,
Ctra Algete km 8, 28110
Algete (Madrid),
SPAIN
IDT Biologika GmbH Ralf Drrwald, Biologika ralf.duerrwald@idt-biologika.de
GmbH, Am Pharmapark,
06861 Dessau-Rosslau,
GERMANY
Finnish Food Safety Authority Anita Huovilainen, anita.huovilainen@evira.fi
EVIRA Officer Finnish Food
Safety Authority EVIRA,
Veterinary Virology,
Mustialankatu 3,
FI-00790 Helsinki,
FINLAND
Kimron Veterinary Institute Hagai Yadin, Head of hagaiy@moag.gov.il
Virology Department and
FMD Laboratory, Kimron
Veterinary Institute,
1 Kimron Street, Rishon
LTzion, ISRAEL
Central Agricultural Office dm Dan, Head of danadam.hu@gmail.com
Laboratory, Central
Agricultural Office,
Veterinary Diagnostic
Directorate, Molecular
Biology Laboratory
Budapest
Tbornok u. 2.
1149
HUNGARY
Stichting Dienst Landbouwkun- Willie Loeffen, Project Willie.Loeffen@wur.nl
dig Onderzoek, part: Central Leader Classical Swine
Veterinary Institute of Wagen- Fever and Aujeskys
ingen UR Disease Central Veterinary
Institute of Wageningen
UR (CVI-Lelystad)
NETHERLANDS

University of Thessaly Charalambos Billinis, billinis@vet.uth.gr
Professor of Virology and
Viral Diseases, Labora-
tory of Microbiology and
Parasitology, Faculty of
Veterinary Medicine Uni-
versity of Thessaly, 224,
43100 Trikalon Karditsa
GREECE
Genome Research Limited, Paul Kellam, Virus pk5@sanger.ac.uk
operating as, Wellcome Trust Genomics Team Leader
Sanger Institute Wellcome Trust Sanger
Institute, Wellcome Trust
Genome Campus, Hinxton,
Cambridge, CB10 1SA,
UNITED KINGDOM
University of Cambridge Derek Smith, Professor of dsmith@zoocam.ac.uk
Infectious disease infor-
matics, Centre for patho-
genic evolution, Down-
ing Street, Cambridge,
CB2 1TN,
UNITED KINGDOM
The Chancellor, Masters and Oliver Pybus, oliver.pybus@zoo.ox.ac.uk
Scholars of the University of Department of Zoology,
Oxford South Parks Road, Oxford,
OX1 3PS,
UNITED KINGDOM
Federal Institute Martin Beer Bundes- martin.beer@fli.bund.de
for Animal Health, forschungssinstitut Fuer
Friedrich-Loeffler-Institute Tiergesundheit, 10 Sude-
ter, 17493 Greifswald-
Insel Riems,
GERMANY
Istituto Zooprofilattico Speri- Giovanni Cattoli, gcattoli@izsvenezie.it
mentale delle Venezie Viale DellUnicersita,
35020 Legnaro,
ITALY
United States Department of Amy Vincent, Veterinary amy.vincent@ars.usda
Agriculture Medical Officer, National
Animal Disease Center,
1920 Daytona Avenue,
Ames 5010,
UNITED STATES
Harbin Veterinary Research Hualan Chen, Director of Hlchen1@yahoo.com
Institute Animal influenza labora-
tory of MOA, 427 Maduan
Street, Harbin 150001
CHINA

Merial S.A.S. Michel Bublot, Virology Michel.Bublot@merial.com
Department, Discovery
Research, Merial,
254 rue Marcel Mrieux,
69007 Lyon
FRANCE
Laboratorios Hipra, S.A. Jaime Maldonado Garcia, jmg@hipra.com
Veterinary Diagnostic
Services DIAGNOS,
Laboratorios HIPRA S.A.,
Avenida La Selva s/n,
Amer 17170 Gerona
SPAIN
Animal Health Trust Debra Elton, Science Pro- debra.elton@aht.org.uk
gramme Manager, Centre
for preventative medicine,
Lanwades Park, Kentford,
Suffolk
CB8 7UU
UNITED KINGDOM
Agrifood and Biosciences Michael Welsh, Head of michael.welsh@afbini.gov.uk
Institute Virology Branch, 18a
New Forge Lane, Belfast,
BT4 3SD,
UNITED KINGDOM
[FLUPIG]
Pathogenesis and transmission

of influenza in pigs
Summary origin, and have receptors for both avian
Pandemic influenza viruses come from wild and mammalian origin viruses. As such, Acronym:
FLUPIG
birds, but must adapt to efficient replica- they represent an ideal vessel for viral
tion and transmission in humans to cause reassortment or adaptation. However, there Project number:
a pandemic. Pigs are considered important is no direct evidence that pigs played a role 258084
intermediate hosts in which avian viruses in any of the three pandemics of the 20th EC contribution:
can adapt to mammals before they trans- century. Only the 2009 pandemic (H1N1) EUR 4 854 452
mit to humans. However, the exact role influenza virus almost certainly comes from Duration:
of the pig and the nature of the genetic pigs. It is still unclear why and how this 54 months
changes required for efficient replication novel H1N1 virus obtained the capacity for Start date:
of avian viruses in pigs and for subsequent human-to-human spread. It is also unclear 1 July 2010
virus transmission between pigs, from what role is played by genes of avian origin
Instrument:
pigs to humans and between humans is in the generation of pandemic viruses, par- collaborative project
still largely unclear. FLUPIG aims to study ticularly in view of the widespread infection
the role of adaptive mutations, genetic of poultry with H5N1 and H9N2 viruses.
reassortment, host and environmental
factors in the adaptation. Another aspect in the influenza ecology is
that pigs support the endemic circulation of
The occurrence and severity of a pan- viruses that belong to the same subtype as
demic also depends on the immune sta- viruses that have been shown (to date) to
tus of the human population. FLUPIG will cause pandemics in humans. Understanding
study the extent and mechanism of cross- how the immune system of pigs responds
protection between antigenically differ- to infection and the extent of cross-protec-
ent H1N1 influenza viruses (heterovariant tion within and between virus subtypes is
cross-protection), and between influenza crucial to limiting the degree of circulation
viruses belonging to different haemagglu- of these viruses in pigs, as is the develop-
tinin subtypes (heterosubtypic cross-pro- ment of novel vaccination strategies. Cross-
tection). We will also evaluate the capac- protection studies in both pigs and ferrets
ity of novel generation vaccines to broaden will also help us to understand the suscep-
cross-protection. tibility of humans to swine-origin influenza
viruses.
Problem
The mechanisms by which influenza viruses Aim
gain the capacity to abandon the ani- We aim at gaining new insights into the role
mal reservoir and become widespread in of pigs in overall influenza ecology, with
human beings are largely unknown. The pig particular reference to the generation of
is believed to play an essential role since human pandemic viruses. In order to allow
pigs are susceptible to all subtypes of influ- us to more accurately predict, respond to,
enza A viruses, including those of avian and control such events, in-depth research
on the pathogenesis and the transmission Potential applications

of influenza viruses between pigs and from Various approaches will be used to design
pigs to other relevant species is essential. and test experimental live-attenuated
In particular, we want to improve our know influenza virus vaccines. The results of
ledge on the gene constellation and genetic these studies will allow us to conclude on
interactions that are necessary to generate a rational vaccine design. Furthermore,
pandemic viruses in combination with an a large collection of genetically modified
improved understanding of host-dependent influenza virus mutants will be generated.
variables such as receptor distribution and The primary goal is to use these mutants
immune response. Combined with improved to study the effect of certain mutations on
surveillance for influenza in animals, pathogenesis and transmission of influenza
effective vaccines and antiviral drugs, this virus. However, certain mutants may turn
knowledge will be critical to the control of out to be potential vaccine candidates.
future influenza pandemics.
Project website
Expected results http://www.flupig.ugent.be/index.html
The use of state-of-the-art technologies
will allow us to develop advanced and Keywords
innovative knowledge on virus-host inter- pig, influenza, pathogenesis, transmission,
actions, specific factors that determine cross protection, pandemic, H1N1, genetic
species barriers and replication efficiency adaptation
of influenza viruses of various origin, and
immune mechanisms that generate protec- Coordinator
tion against homologous and heterologous Prof. Kristien Van Reeth
influenza virus subtypes. This knowledge is Universiteit Gent
of critical importance to assess the prob- Salisburylaan 133
ability and risk of transmissibility of influ- 9820 Merelbeke
enza viruses from swine to other mam- BELGIUM
malian hosts, and further spread within flupig@ugent.be, kristien.vanreeth@ugent.
mammalian hosts. be
The knowledge generated by FLUPIG will

provide clear insight into the role of pigs in
overall influenza ecology and, in particular,
in the zoonotic potential of SIVs. Conse-
quently, it will allow us to improve preven-
tion and control strategies for human influ-
enza pandemics.
Partners
CONTACT ORGANISATION
Dr Ilaria Capua Istituto Zooprofilattico Sperimentale delle Venezie,
Legnaro, ITALY
Prof. Wendy S. Barclay Imperial College London, London, UNITED KINGDOM
Prof. Ian H. Brown Veterinary Laboratories Agency, London,
UNITED KINGDOM
Dr Mikhail Matrosovich Philipps Universitt Marburg, Marburg, GERMANY
Prof. Guus F. Rimmelzwaan Erasmus Medisch Centrum, Rotterdam, NETHERLANDS
Prof. Iwona Markowska-Daniel Panstwowy Instytut WeterynaryjnyPanstwowy Instytut
Badawczy, Pulawy, POLAND
Prof. Malik Peiris HKU Pasteur Research Centre, HONG KONG
Prof. Jrgen A. Richt Kansas State University, Manhattan, Kansas,
UNITED STATES
Prof. Thomas Mettenleiter Friedrich-Loeffler-Institut, Greifswald-Insel Riems,
GERMANY
European Commission
Luxembourg: Publications Office of the European Union
2012 213 pp. 17.6 x 25.0 cm
ISBN 978-92-79-21035-8
doi:10.2777/73975
How to obtain EU publications
Free publications:
This publication is dedicated to the memory of Isabel Minguez
via EU Bookshop (http://bookshop.europa.eu);
Tudela who made a major contribution to the Animal Health at the European Unions representations or delegations. You can obtain their contact details on the
research sector over many years in the Commission services, but Internet (http://ec.europa.eu) or by sending a fax to +352 2929-42758.
who sadly passed away on 16 April 2011.
Priced publications:
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and reports of cases before the Court of Justice of the European Union):
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EUROPEAN COMMISSION

Directorate E Biotechnologies, Agriculture, Food
Unit E4 Agriculture, Forest, Fisheries, Aquaculture
E-mail: RTD-FP7-KBBE-Activity1@ec.europa.eu
Contact: Anne-Sophie Lequarre
European Commission
Office SDME 08/018
B-1049 Brussels
Tel. (32-2) 29-80068

Fax (32-2) 29-63029
E-mail: Anne-Sophie.Lequarre@ec.europa.eu
KI-31-11-002-EN-C
This publication gathers information on initiatives
funded by the European Commission over the last
decade regarding animal health challenges. There are
Animal Health

some general actions which have been developed for
the harmonisation of European funding schemes, for
the prioritization of research topics or for implementing
improved surveillance systems on emerging and re-
emerging diseases. Several projects concern notifiable
diseases, which are often highly epidemic and are
Research
frequently regulated by the culling of infected animals.
The goal pursued is to achieve the capacity to react more
quickly by using fast and reliable diagnostics tools, but
also to develop alternative disease eradication protocols
through new generation vaccines. Regarding endemic
diseases, which decrease the efficiency and profitability
of the livestock sector and are
difficult to phase out, the aim is to progressively reduce
their incidence. Finally, there is a chapter which gives
information on projects targeting the influenza viruses,
for controlling animal infection and improving our
knowledge on cross-species infection within a one
health vision.
Research and
Innovation

A Decade of EU-funded Animal Health

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A decade of EU-funded Animal Health Research

Directorate-General for Research and Innovation

Contact: Anne-Sophie Lequarre

Tel. (32-2) 29-80068

FP7 Cooperation Work Programme

Directorate-General for Research and Innovation

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CHAPTER 1. NETWORK, SURVEILLANCE  11

[EMIDA] Coordination of European research on emerging

[STAR-IDAZ] Global Strategic Alliances for the Coordination of Research

[EPIZONE] Network of Excellence

[DISCONTOOLS] Development of the most effective tools

[Arbo-Zoonet] Network for Capacity-building

[WildTech] Novel Technologies for Surveillance of Emerging

[ICONZ] Integrated control of neglected zoonoses:

CHAPTER 2. EPIDEMIC DISEASES OF LIVESTOCK  41

European policies for animal health sustained

2.1 Foot-and-mouth disease  44

[FMD_ImproCon] Improvement of foot-and-mouth disease control

[CA FMD/CSF] Foot-and-mouth disease (FMD)

[FMD-Disconvac] Development, enhancement and complementation

2.2. Classic swine fever  59

[CSFVACCINE & WILD BOAR] Epidemiology and control

[CSF-GoDIVA] Improve tools and strategies for the prevention

2.3. African swine fever  71

[ASFRISK] Evaluating and controlling the risk

[BTVAC] Improved vaccines for bluetongue disease  77

[Medreonet] Surveillance network of reoviruses, bluetongue

[ORBIVAC] Development of vaccines for bluetongue virus (BTV),

CHAPTER 3. ENDEMIC DISEASES 95

The economic impact and research synergy resulting

3.1. Mycobacterial diseases  101

[VENoMYC] Veterinary network of laboratories researching

[TB-STEP] Strategies for the eradication of bovine tuberculosis  107

3.2. Parasites  112

[DELIVER] Design of effective and sustainable control strategies

[PARASOL] Novel solutions for the sustainable control

[PARAVAC] Vaccines against helminth parasites of livestock

3.3. Tick-borne diseases  125

[PiroVac] Improvement of current and development of new vaccines

3.4. Porcine circovirus diseases  129

[PCVD] Control of porcine circovirus diseases (PCVD):

[NMSACC-PCVD] PCVD: Towards improved food quality and safety

3.5. Porcine reproductive and respiratory syndrome  141

[PoRRSCon] New tools and approaches to control Porcine Reproductive

CHAPTER 4. INFLUENZA  147

Introduction by Ilaria Capua 148

4.1. Avian flu 150

[AIV-VACC] Vaccine, diagnostic test development

[FLUAID] Generation of information and tools to support the management

[Healthy Poultry] Development of new integrated strategies

[FLUPATH] Avian influenza: Impact of virus-host interactions

[New-Flubird] Network for early warning of influenza viruses

[FLURESIST] Avian influenza virus survival in poultry commodities,

[Rivers] Resistance of influenza viruses in environmental reservoirs

[FLUTEST] Improved diagnosis and early warning systems

CHAPTER 1. NETWORK, SURVEILLANCE 11

CHAPTER 2. EPIDEMIC DISEASES OF LIVESTOCK 41

2.1 Foot-and-mouth disease 44

2.2. Classic swine fever 59

2.3. African swine fever 71

[BTVAC] Improved vaccines for bluetongue disease 77

CHAPTER 3. ENDEMIC DISEASES 95

3.1. Mycobacterial diseases 101

[TB-STEP] Strategies for the eradication of bovine tuberculosis 107

3.2. Parasites 112

3.3. Tick-borne diseases 125

3.4. Porcine circovirus diseases 129

3.5. Porcine reproductive and respiratory syndrome 141

CHAPTER 4. INFLUENZA 147

Introduction by Ilaria Capua 148

4.1. Avian flu 150

4.2. Swine flu 202

[ESNIP 2] European Surveillance Network for Influenza in Pigs 2 202

[ESNIP 3] European Surveillance Network for Influenza in Pigs 3 204

[FLUPIG] Pathogenesis and transmission of influenza in Pigs 211

2. The European effects will be mirrored

3. The project results will also have indi-

of distinguishing vaccinated and infected 6. Important data on the safety of inacti-