AQUACULTURE HEALTH I N T E R N A T I O N A L

ISSUE 13 JUNE 2008 NZ$10.00

NEW CALEDONIA BREAKS

SHRIMP INBREEDING

ALIEN FISH INDICATE

AQUATIC HEALTH
BIOFLOC HAS POTENTIAL
TO FIGHT INFECTION
CONTENTS ISSUE 13, JUNE 2008

3 EDITORIAL
Finding the key to good biosecurity
4 RESEARCH FOCUS
Breaking inbreeding in domesticated shrimp (Litopenaeus stylirostris) in New
Caledonia
7 NEWS FEATURE
Venezuela passes new law in fisheries and aquaculture
8 RESEARCH FOCUS
Horizon scanning: the potential use of biofloc as an anti-infective strategy in
aquaculture – an overview
11 NEW PUBLICATION
Pearl Oyster Health Management manual on line
12 RESEARCH FOCUS
The BC Pacific Salmon Forum, Nanaimo, Canada
4 15 AQUAFEED
A news roundup courtesy of the Aquafeed website www.aquafeed.com
16 NEWS
Updates from around the globe
22 RESEARCH FOCUS
Selective breeding can reduce salmon lice problems
23 EVENTS
Forthcoming fish and shellfish events
24 COMMERCIAL FOCUS
Cytogenix announces agreement to develop DNA vaccines
28 LABORATORY FOCUS
The Central Fish-Health Laboratory in Israel
32 RESEARCH FOCUS
What’s new in pancreas diseases research?
34 BOOK REVIEW AQUACULTURE
Aquaculture biosecurity: prevention, control
HEALTH I N T E R

22 and eradication of aquatic animal disease NEW CALEDO

N A T I O
ISSUE 13
N A L
JUNE 2008
NZ$1 0.00

NIA BREAKS
36 ORNAMENTALS FOCUS S HRIMP INBREE
DING
A basic overview of multicellular parasitic
infections
38 VACCINE STATISTICS
Monthly Norwegian
vaccine statistics ON THE
provided by Pharmaq COVER ALIEN FISH IND
AQUATIC HEA ICATE
40 RESEARCH FOCUS A farmed shrimp BIOFLOC HA
S
LTH
TO FIGHT INF POTENTIAL
Spread and L stylirosis in New ECTION

colonisation of alien Caledonia is tagged with

fish species in open coloured silicone. .
waters: a reliable MAIN PHOTO COURTESY
36 indicator of aquatic J Patrois, Ifremer
health INSET PHOTO COURTESY
Y Harache, Ifremer

EDITORIAL DIRECTOR: WEBSITE: Web4U

Dr Scott Peddie
CONTRIBUTORS:
ISSN 1176-86330 ISSN (web) 1176-8649 PUBLISHER: D Ansquer, Kartik Baruah, Gerald Bassleer,
Keith Ingram
An informative journal for the Peter Bossier, F Broutoi, P Brun, David Conroy,
aquaculture health professional MANAGER: Vivienne Ingram S de Decker, Suzi Fraser Dominy, R Dufour, Rama

Published by:
VIP PUBLICATIONS LTD
4 Prince Regent Drive
Half Moon Bay, Auckland 1706
New Zealand
Ph +64 9 533 4336, Fax +64 9 533 4337
Email keith@aquaculturehealth.com
www.aquaculturehealth.com
ACCOUNT MANAGER: Fiona Peddie Falk, C Galine, Bjarne Gjerde, E Goyard, C Goarant,
Y Harache, WS Lakra, Marian McLoughlin,
ASSISTANT EDITOR: Mark Barratt-Boyes
J-R Maillez, J Patrois, Scott Peddie, JM Peignon,
DESIGNER: Rachel Walker D Pham, AK Singh, Amit Kumar Sinha, E Vourey
GENERAL: Reproduction of articles and materials published in Aquaculture Health International in whole or part, is permitted, provided the source and author(s)
are acknowledged. However, all photographic material is copyright and written permission to reproduce in any shape or form is required. Contributions of a nature
relevant to the aquaculture industry are welcomed and industry participants are especially encouraged to contribute. Articles and information printed in
Aquaculture Health International do not necessarily reflect the opinions or formal position or the publishers unless otherwise indicated. All material published in
Aquaculture Health International is done so with all due care as regards to accuracy and factual content, however, the publishers cannot accept
responsibility for any errors and omissions which may occur. Aquaculture Health International is produced quarterly.

2 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008

 EDITORIAL
FINDING THE KEY
TO GOOD BIOSECURITY
SCOTT PEDDIE, EDITORIAL DIRECTOR

T
he team at Aquaculture Health International is delighted to bring
you the thirteenth issue of the magazine, celebrating our third
full year of production. As always, we have endeavoured to bring
you a diverse range of articles, and so we have included contributions
from Israel, Ireland, Norway, India, New Caledonia, France, Venezuela,
Belgium, the United States of America and Canada.
This issue of Aquaculture Health International also contains a review
on David Scarfe, Cheng-Sheng Lee and Patricia O’Bryen’s excellent
book entitled Aquaculture Biosecurity. Although this book was first
published in 2006, it is still the most authoritative and comprehensive
publication on this subject that I’ve come across to date.
Clearly, biosecurity is one of the most important issues for policy
makers, regulators, fish health professionals and aquaculturists alike.
You only have to follow the unfolding saga of the current ISA outbreak
in Chile on the news wires to get a sense of the centrality of this issue
to the aquaculture industry.
There is, however, one area that we tend to miss when discussing
biosecurity. Decision makers quite rightly have a tendency to focus
almost exclusively on setting and enforcing norms and standards in
this arena, whether it be on an international, national or regional
platform. Where we often fall short, and this is an area that is touched
on in Scarfe, Lee and O’Bryen’s book, is the issue of the mindset of
those at the “coal face” in the industry, namely the staff who work on
the farms. I’ve visited farms where the biosecurity procedures have
been exemplary on paper but have been half-heartedly adhered to, not
because the staff are unaware of company policy, but rather because
they are “de-motivated” by the environment they are working in. This
is perhaps a more serious issue than many fish health professionals are
aware of, and it’s to the credit of Scarfe, Lee and O’Bryen that they have
included a short chapter on this very topic.
I once again find myself calling for a more holistic approach, as I
often do in these columns, although this time on a different aspect of
aquaculture health management. It seems to me that there is a tendency
to compartmentalise the myriad functions involved in managing an
aquaculture unit so that training in biosecurity and developing a highly
motivated team are seen as having only a tenuous link at best.
The bottom line is that a properly motivated and valued workforce
is much more likely to take on ownership of the biosecurity process,
and this is surely to the advantage of all concerned. After all, sub-
optimal biosecurity has the potential to seriously impact on public
perception of the industry, in addition to a plethora of other negative
impacts of the biological and economic variety.
So I find myself musing on a number of issues. For example, how
can the latest advances in business communication theory be utilised
and developed to inculcate a sense of biosecurity awareness? What are
the motivational models that can best be applied to the aquaculture
environment to enable staff at all levels in the enterprise to promote
and foster “good practice?”
It seems to me that these issues need to be investigated more
thoroughly at both a theoretical and practical level. We need all the
tools we can to foster and promote optimal biosecurity in whichever
sector of the industry we work in. ■

SUBSCRIBE NOW! Be sure to get your copy of Aquaculture Health International direct by email

Name__________________________________________________________________________________________________________________

Address _______________________________________________________________________________________________________________

__________________________________________________________________________________________ Postal code _________________

Email __________________________________________________________________________________________________________________

ENCLOSE A CHEQUE FOR _____________________ NZ$40.00 Electronic version by email, see www.aquaculturehealth.com

■ Visa ■ Mastercard ■ Bankcard (other cards are not accepted)

Card Number __________________________________________________________________________________________________________

Card Name ___________________________________________________________________________________________________________

Signature ________________________________________________________ Expiry date ——— / ———

POST TO: VIP Publications Ltd, 4 Prince Regent Drive, Half Moon Bay, Auckland 1706, New Zealand GST No: 68-684-757

JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 3

RESEARCH FOCUS
BREAKING INBREEDING
IN DOMESTICATED SHRIMP
(LITOPENAEUS STYLIROSTRIS)
IN NEW CALEDONIA
BY E GOYARD, C GOARANT, D ANSQUER, F BROUTOI, P BRUN, S DE DECKER, R DUFOUR, C GALINIE, J-R MAILLEZ,
JM PEIGNON, D PHAM, E VOUREY, J PATROIS AND Y HARACHE
IFREMER, DEPARTEMENT AQUACULTURE EN CALÉDONIE, NOUMÉA CEDEX, NEW CALEDONIA
SODACAL, A 133HA INDUSTRIAL FARM
CREATED IN 1983. PRODUCTION RANGES
FROM 320 TONNES TO 490 TONNES
PHOTO CREDIT: Y HARACHE – IFREMER
PHOTO CREDIT: Y HARACHE – IFREMER
I
n 1993, when temperatures dropped rapidly at the beginning of
the cold water season, significant mortalities occurred among
the farmed shrimp (Litopenaeus stylirostris) population in New
Caledonia. The mortalities, due to Vibrio penaeicida, were locally
referred to as “1993 syndrome” and within a few months they had
spread to all the farms on the island.
Survival rates (pl’s to commercial size shrimps) declined
repetitively down to 30-35 percent during the winter, making winter
production uneconomical. The industry concentrated on seasonal
production during the warm months, with good results, although
such an approach created bottlenecks for market availability and
peak demand of post-larvae over short periods.
At the end of 1997, another sanitary crisis appeared during the
warm season in the same farm initially struck with V penaeicida.
This was due to a septicaemic summer vibriosis caused by Vibrio
nigripulchritudo, which has occurred every year since on the
contaminated site. V nigripulchritudo (both non-pathogenic and
highly pathogenic strains) was also isolated in several other sites, but
was not always associated with mortalities.
While V penaeicida isolates show a low genetic variability, the strains
of V nigripuchritudo appear highly variable, with striking differences
in their pathogenicity level. As New Caledonia’s production certified
procedure totally prohibits the use of antibiotics in the pond-
growing phase, the presence of this pathogen was considered a major
potential threat to industry expansion.
This motivated a specific monitoring policy, undertaken by
Ifremer in collaboration with DAVAR, the Nouvelle-Calédonie
Veterinary Service. Significant summer mortalities, due to the same
pathogen, appeared in 2003 and 2005 in two other farms. However,
production continued to increase slowly, reaching 2430 tonnes in
4 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
STYLIROSTRIS
2006, with particularly strong exports markets in France, Japan and
Australia.
The domesticated strain, introduced into the country in the
late 1970s and reproduced in captivity since, shows good growth
performance profiles and an excellent virus status profile (resistant
to IHHNV and free of all other known viruses).
However, we were convinced that the low number of founders had
induced a high level of inbreeding with an important loss of genetic
variability, possibly hampering its adaptability to modifications of
the rearing conditions, including resistance to new pathogens.
The results of the multi-disciplinary Ifremer DéSans programme
pointed out that the outbreaks of seasonal diseases were linked to
the instability of the pond ecosystem (sediment and water column)
during intensified production cycles, the virulence of two specific
pathogens (Vibrio nigripulchritudo or V penaeicida, respectively) and
the shrimps’ capacity to resist the stress induced by the interaction
of these critical factors.
We assumed there was a genetic component to this global
resistance capacity. The necessity to re-introduce “new blood” into the
broodstock line, even through a costly, complex and risky procedure,
appeared unavoidable. This provided the impetus for a range of
disease challenge studies using stocks of different genetic origins.
STOCKS TESTED
A domesticated SPF strain from Hawaii (High Health Aquaculture
Ltd) was purchased by the New Caledonian Industry (UPRAC
association), allowing it to test three discrete stocks for their
performance:
• CC. New Caledonian stock from Ifremer and private hatcheries. A
newly constituted CC control line (to reproduce every year) was

PHOTO CREDIT: Y HARACHE – IFREMER
HAWAII SHRIMPS IN
QUARANTINE AT UPRAC
PHOTO CREDIT: J PATROIS – IFREMER
COLOUR SILICONE TAGGING
OF HAWAII SHRIMP FAMILIES
created with 16 bi-parental families and reproduced twice in 2005
and 2006.
• HH. Hawaiian stock introduced through a quarantine procedure
(UPRAC) was reproduced at Ifremer, constituting a new HH-
G9 and HH-G10 generations structured each in at least 16 bi-
parental families from known genealogy, reducing inbreeding to
a minimum. From this G9, batches of juveniles for experimental
evaluation of the Hawaii stock were constituted in 2005 and 2006.
• Hybrid-F1. HH and CC animals were crossed each year to obtain
two first-generation hybrid stocks named F1-2005 and F1-2006
for evaluation of performance, using several families for each
combination sex/origin.
EVALUATION OF THE COMPARATIVE
PERFORMANCES OF THE DIFFERENT
POPULATIONS: PRELIMINARY RESULTS
Winter tests in earthen ponds (Ifremer 2006 and 2007)
The different groups arising from 2005 (CC-2005, HH-2005 and
F1-2005) and 2006 reproduction scenarios (CC-2006, HH-2006
and F1-2006) were reared separately in earthen ponds at the Ifremer
station. They were then branded with silicone injections (2.9g to 11g
mean weight) and re-mixed in two 500m2 ponds seeded with 10,000
shrimp and reared for 130 days using normal production techniques.
The ponds were harvested after the beginning of the cold season,
with identification, counting and size sampling of all groups.
Under these “competitive” conditions, the average results showed,
for both years, a higher survival rate for Hybrid F1 (respectively
60.5 and 53.5 percent) compared to HH (60 and 32.5 percent) and
CC (51 and 34.5 percent). Although no moribund shrimp could be
observed, V penaeicida was not isolated, but the presence of a few
THE IFREMER RESEARCH
STATION AT SAINT VINCEN
PHOTO CREDIT: Y HARACHE – IFREMER
PHOTO CREDIT: J PATROIS – IFREMER
EXPERIMENTAL FLOATING CAGES ON SITE AT THE
AIGUE-MARINE FARM AFFECTED BY OUTBREAKS
OF SUMMER VIBRIOSIS
dead shrimp found following rapidly dropping temperatures was
typical of syndrome 93, suggesting a higher resistance by Hybrids-F1
to V penaeicida. The final weight, and the subsequent growth rate,
were significantly higher for Hybrid F1 (0.22g and 0.24g/day) than
for both parent HH and CC lines (0.15g to 0.17g/day).
Summer tests in floating cages (Private farm 2007)
Because no summer mortalities caused by V nigripulchritudo
had ever been observed in the Ifremer ponds, it was decided, in
coordination with the Farmers Association, to compare the results
of the three populations in a farm showing typical outbreaks of V
nigripulchritudo since 2003.
This test was conducted in small floating cages (4m2/3.2m3) installed
in a production pond affected by summer mortalities. Shrimps were
stocked either as “pure line” cages or under “competition” (mixed
population) during the recurrent outbreak period.
The final survival after 77 days appeared significantly higher in
Hybrids F1 (51 percent) than the parent lines, which expressed no
significant difference between CC and HH (41 and 44 percent) with
no significant difference between the conditions of testing (“pure
line” cages vs “competition” cages).
Haemolymph samples taken at day 29 showed a V nigripulchritudo
prevalence of 90 to 100 percent with no difference between groups,
but a higher number of vibrios in CC samples (50 percent with more
than 300 vibrios in 10µl) while 50 percent of HH and F1 bore fewer
than 50 pathogens. Further samples did not indicate significant
trends in prevalence between groups, but confirmed a higher number
of V nigripulchritudo CFUs found in CC haemolymph samples.
The average growth rates observed in Hybrids F1 were 40
percent higher than that of the parent lines (0.28g v 0.19g/day),
JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 5
RESEARCH FOCUS
DISEASE CHALLENGE STUDIES OF INBRED AND OUTBRED SHRIMP IN NEW CALEDONIA

EXPERIMENTAL STRUCTURES AND PROGRAM FUNDED THOUGH A 5 YEARS

PHOTO CREDIT: J PATROIS – IFREMER

“CONTRAT DE DÉVELOPPEMENT” ESTABLISHED BETWEEN THE STATE OF FRANCE
AND THE PROVINCES OF NEW-CALEDONIA

THE EXPERIMENTAL INFECTION ROOM FOR

STANDARDISED TEST RESISTANCE TO VIBRIO

IFREMER STAFF AT SAINT VINCENT, MAY 2007 (PERMANENT STAFF

IFREMER SHRIMP PONDS
CONTRIBUTING FROM 2002 TO 2006) D ANSQUER, P BOISARD, O BOUISSOU,
F BROUTOI, P BRUN, L CHIM, D COATANEA, L DELLA-PATRONA, R DUFOUR, E
GOYARD, C GOARANT, L GOURMELEN, Y HARACHE, A HERBLAND, J HERLIN,
JS LAM, C LAMBERT, P LEMAIRE, H LEMONNIER, JR MAILLIEZ, AL MARTEAU,
A MATEAU, H MICHAUT, C MUGNIER, J PATROIS, JM PEIGNON, J PICHON, D
PHAM, E PITA, JM RANOUIL †, E SAULNIER, BG SERY, SOULARD, N WABETE, N
WAHMETRUA, B WAPOTRO AND K WASSAUMII

TEMPORARY STAFF: M CASTEX, C JUSTOU, A LEGRAND, R LUCAS, S DE DECKER,

F IMBERT, E VOUREY AND E ZIPPER.

INPUTS FROM OTHER IFREMER LABORATORIES OR EXTERNAL PARTNERS: E

BACHÈRE, C COURTIES, C CAHU, G CUZON, J DE LORGERYL, C GALINIÉ,
F LEROUX, AG MARTIN, JL MARTIN, JC MASSABUAU, D SAULNIER, S VIRLY.
ESTABLISHED PARTNERSHIPS WITH IRD, INSTITUT PASTEUR, UNIVERSITÉ DE
NOUVELLE-CALÉDONIE, DAVAR (VETERINARY SERVICES OF NOUVELL-
CALÉDONIE, UNIVERSITÉ DE BORDEAUX ET PARIS VI-BANYULS)

FIGURE 1: Relative performances of Hybrid-F1 population (A) and

of the Hawaiian population (B) compared with the Caledonian
population

Production of Biomass in ponds - year 1
Growth in ponds - year 1
Survival in ponds - year 1
240%
Production of Biomass in ponds - year 2 Production of Biomass in cages with a much better food conversion ratio than the CC line in the
Growth in ponds - year 2 Growth in cages
testing conditions, though this remains to be confirmed in other
Survival in ponds - year 2 Survival in cages
conditions.

220%
A Experimental infections with Vibrio penaeicida (Ifremer 2007)
Six experimental infection tests were undertaken, three each in 2006
200% and 2007, in a bio-safe infection room at Ifremer. The experimental
180%
population comprising the three tagged populations (CC, HH and
Hybrid F1) was reared in an outdoor pond for three weeks. Groups
160% of each population were then introduced in the experimental facility
in 16 or 32 x 200-litre tanks. After acclimatisation for one week, the
140%
animals were infected by balneation or injection with a reference
120% strain of V penaeicida, following a standardised technique.
Natural contamination in 2007 with V nigripulchritudo prior
100%
30% 35% 40% 45% 50% 55% 60% to the V penaeicida experimental infections made the expression
Biomass production in ponds - year 1 Biomass production in ponds - year 2 Biomass production in cages
of results and the characterisation of a specific resistance to V
Growth rate in ponds - year 1 Growth rate in ponds - year 2 Growth rate in cages penaeicida difficult. However, in all experiments, the final survival
Survival in ponds - year 1 Survival in ponds - year 2 Survival in cages
of hybrids F1 was higher than that of CC and HH, with a significant
140% difference for 2006 experiments (57 percent v 50 and 47 percent
B respectively).
120%

100% CONCLUSION
80%
All the tests indicate that F1 hybrids are characterised by faster
growth, independently of the testing conditions: in earthen ponds,
60%
in cages, under competition with other strains or alone; while the
40% slower-growing pure strains CC and HH do not show significant
20%
differences.
Though the present results do not allow a conclusion to be drawn
0%
30% 35% 40% 45% 50% 55% 60% with respect to a specific resistance of the hybrids to the pathogenic
Vibrios, it was shown in all experiments that they provide higher
Average values for survival, growth rate and biomass, obtained in
ponds and cages, is expressed in % of the values observed in survival than any parent population in all cases (pond or cage
Caledonian shrimps (CC) for the same criteria. (Goyard et al., 2008) rearing, winter or summer period and global survival to experimental
handling) and infections. Moreover, indications of a lower pathogen

6 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008


VENEZUELA PASSES NEW LAW IN
FISHERIES AND AQUACULTURE
SOURCE: DA CONROY PHD, CBIOL, FIBIOL, EMERITUS PROFESSOR OF FISH PATHOLOGY,
FACULTY OF VETERINARY SCIENCES, CENTRAL UNIVERSITY OF VENEZUELA, MARACAY, VENEZUELA
T
he government of Venezuela passed a new Fisheries and
Aquaculture Law on March 14 which will have a considerable
impact on the future development of its fisheries and
aquaculture.
The new law, known as Ley de Pesca y Acuicultura, published as
Decree Nº 5930 in the Official Gazette, or Gaceta Oficial de la República
Bolivariana de Venezuela (GORBV), Nº 5877 (extraordinario), on
March 14, is very extensive in its coverage and attributions, as much
with reference to “fisheries” as to “aquaculture”.
It specifically replaces and leaves without any legal effects the
previous laws on fisheries and aquaculture, published in the GORBV
Nº 37323 of November 13, 2001, and in the GORBV Nº 37726 of
May 22, 2003, respectively.
The National Institute of Fisheries and Aquaculture (Instituto
Nacional de Pesca y Acuicultura, or INAPESCA), opportunely
created by these previous two laws as a dependency of the Ministry
of Agriculture and Lands, or MAT, is now officially designated as the
Socialist Institute of Fisheries and Aquaculture (Instituto Socialista
de Pesca y Acuicultura, or INSOPESCA), which continues to be a
dependency of the MAT.
The printed version of this new law comprises some 120 pages
of text, and its contents should be read very carefully by all those
with an interest in aquaculture in Venezuela (preferably with some
professional input from the interested parties’ financial, legal and
technical advisers, where convenient and/or necessary).
The law provides a series of quite clear definitions of the various
types of aquaculture activities to which it refers, eg
• freshwater and marine
• extensive, semi-intensive and intensive, including ponds, tanks,
▲
CONTINUED FROM PAGE 30
load in haemolymph during asymptomatic infections than any of
the parent lines were obtained.
Although full-scale testing in commercial ponds is essential to reach
a final conclusion, the combined average improvements in growth
and survival inferred from all experiments would provide a biomass
gain ranging from 60 to 130 percent in the historic Caledonian strain,
while no difference would occur between parent lines.
SELECTED BIBLIOGRAPHY
Goarant C, Ansquer D, Herlin J, Domalain D, Imbert F and De Decker
S 2006. “Summer syndrome” in Litopenaeus stylirostris in New
Caledonia: Pathology and epidemiology of the etiological agent,
Vibrio nigripulchritudo. Aquaculture 253. pp105-113
Goarant C, Reynaud Y, Ansquer D, de Decker S, Saulnier D and
Le Roux F 2006. Virulence and molecular epidemiology of Vibrio
nigripulchritudo, a pathogen for cultured penaeid shrimp (Litopenaeus
stylirostris) in New Caledonia. Systematic and Applied Microbiology
29. pp570-580
Goarant C, Merien F 2006. Quantification of Vibrio penaeicida,
the etiological agent of Syndrome 93 in New Caledonian shrimp
farming, by real-time PCR using SYBR Green I chemistry. Journal
NEWS FEATURE
floating cages and pens
• algae, crustaceans, molluscs and finfish, and their larval stages
where appropriate
• aspects of aquatic animal health, including quarantine matters
and genetics
and the levels of these activities (eg, artesanal, small-scale operations,
larger commercial operations). Tariffs are established for issuing
various types of permits, certificates, authorisations and other such
“documentation” on a sliding scale that favours artesanal producers
and becomes successively more “onerous” for the larger producers.
In compliance with the terms of article 47 of Title VI (Communal
Responsibility) of the law, for example, aquaculture producers at all
levels are now obliged to freely donate, directly and gratuitously to
government and local community organisations a minimum of five
percent of their production for distribution by those agencies to
needy sectors of the population.
This article also states quite clearly that the competent authorities
can increase this free contribution of five percent as and when
deemed necessary.
In accordance with the general guidelines as detailed in the text
of this new Fisheries and Aquaculture Law, the future development
of aquaculture in Venezuela is to be oriented above all else toward
improving the “social welfare of the less economically favoured
communities” within the country.
The contents of this law should be carefully read and studied by
any interested parties. Potential investors, in particular, would be
strongly recommended to adopt a "caveat emptor" approach before
reaching any important decisions in principle. ■
of Microbiological Methods 67. pp25-27
Goyard E, Goarant C, Ansquer D, Brun P, de Decker S, Dufour R, Galinié
C, Peignon JM, Pham D, Vourey E, Harache Y and Patrois J 2008. P.
Cross-breeding of different domesticated lines as a simple way for
genetic improvement in small aquaculture industries: Heterosis and
inbreeding effects on growth and survival rates of the Pacific blue
shrimp Penaeus (Litopenaeus) stylirostris. Aquaculture 275. pp43-50
Harache Y, Herbland A 2004. Le programme DéSanS (Défi Santé
Stylirostris) appliqué à la filière crevette calédonienne. Trente ans
de cerevetticulture en Nouvelle-Calédonie. Nouméa-Koné. June 2-6,
2003. (ed) Ifremer, Actes Colloq, 38, pp280.
Saulnier D, Haffner P, Goarant C, Levy P, Ansquer D 2000. Experimental
infection models for shrimp vibriosis studies: a review. Aquaculture
191. pp133-144
Patrois J, Goarant C, Goyard E, Harache Y, Primot P and Bador R
2007b. Blue shrimp quarantined in New Caledonia; genetic variability
programme. Global Aquaculture Advocate 10 (5). pp90-92
CONTACT DETAILS
Ifremer, Departement Aquaculture en Calédonie, BP 2059, 98846
Nouméa Cedex, New Caledonia. Contacts: Emmanuel.Goyard@
ifremer.fr or harache.aqua@orange.fr ■
JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 7
RESEARCH FOCUS
HORIZON SCANNING: THE
POTENTIAL USE OF BIOFLOC AS
AN ANTI-INFECTIVE STRATEGY IN
AQUACULTURE – AN OVERVIEW
BY AMIT KUMAR SINHA, KARTIK BARUAH AND PETER BOSSIER
LABORATORY OF AQUACULTURE AND ARTEMIA REFERENCE CENTRE, DEPARTMENT OF ANIMAL PRODUCTION,
GHENT UNIVERSITY, GHENT, BELGIUM
I
ntensification of the aquaculture industry has in some cases been
accompanied by environmental pollution (high nitrogen toxicity)
and increasing severity of disease outbreaks. The increased level
of intensification requires more supplementary protein feed and in
turn contributes to nitrogen pollution in the culture system.
Decomposition of dead algae, uneaten feed, fish excreta and
other organic matter releases excess nitrogen in the form of
ammonia and nitrite and amplifies the nitrogen toxicity level.
Elevated concentrations of ammonia affect growth, moulting (in
shellfish), oxygen consumption and even ammonia excretion, and
can eventually cause mortality of fish/shellfish. Increased ambient
nitrite concentration negatively affects the growth performance
and survival of fish/shellfish (Mallasen and Valenti 2006). Hitherto,
many techniques such as photo-autotrophic algae-based systems
(green-water systems), frequent water exchange and the use of
biofilters have been used to remove excessive nitrogen.
However, none of these options has proved to be technically
feasible and economically viable. Thus, the quest to develop an
economical, eco-friendly and user-friendly strategy to remove or
optimise the level of such toxic compounds in the pond water has
become a priority.
The breakdown of PHA inside
the gastrointestinal tract can be
carried out via enzymatic and
chemical hydrolysis
One such strategy for removing ammonium nitrogen from
water is through its assimilation into microbial proteins by adding
carbonaceous material into the system. Nitrogenous compounds
present in the water combine covalently with carbon and produce
flocs of protein molecules. These small protein molecules flocculated
together form a floating mass called bioflocs; these flocs are
dominated by bacteria. An important aspect of this process is the
potential utilisation of microbial protein as a source of protein
for fish and shrimps. The success of this technique depends upon
optimising the amount of carbonaceous material to be added.
Recently, it was observed that excess addition of the carbon
source enhances the development of polyhydroxyalkanoate (PHA)
accumulating micro-organisms. The properties of PHAs are very
similar to those of organic acid and have been proved to be effective
bio-control agents, given that they beneficially affect the host’s
microbial balance in the gut.
8 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
PRINCIPLE
Bacteria and other micro-organisms require carbohydrates for their
growth. As protein is the major component of new cell material,
nitrogen is also required as an essential element. Thus, microbial
utilisation of carbohydrate or any other low nitrogen feed is
accompanied by the immobilisation of inorganic nitrogen. This is
the basic process carried out by the microbial community.
BIOFLOC AND WATER QUALITY
IMPROVEMENT
Nitrogen control is stimulated by feeding bacteria with carbohydrates,
and through the subsequent uptake of nitrogen from the water, by
the synthesis of microbial protein. The relationship between adding
carbohydrates, reducing ammonium and producing microbial
proteins depends on the microbial conversion coefficient, the C/N
ratio in the microbial biomass and the carbon contents of the added
material (Avnimelech 1999).
The C/N ratio has been widely used as an index of the rate at which
organic matter decomposes. If the organic matter is low in nitrogen
content (ie a high C/N ratio), some of the nitrogen for microbial
growth will be obtained from the water column and will become
immobilised as microbial protein. Previous studies have shown
that the immobilisation of inorganic nitrogen only occurs when
the C/N ratio of the organic matter is higher than 10 (Lancelot and
Billen 1985). However, Avnimelech (1999) verified that to eliminate
inorganic nitrogen, the C/N ratio in pond should be 10.75, rising to
15.78 when fish are introduced into the holding pond.
BIOFLOC AS FISH NUTRIENTS
The bacterial protein and new cells (single-cell protein)
synthesised by the heterotrophic bacterial population are utilised
directly as a food source by the cultured organisms (carp,
tilapia, shrimp), thus lowering the demand for supplemental
feed protein (Avnimelech 1999). Hari et al (2004) reported
that Penaeus monodon could effectively utilise the additional
protein derived from the increased bacterial biomass as a result
of carbohydrate addition. Burford et al (2004) suggested that
“flocculated particles” rich in bacteria and phytoplankton could
contribute substantially to the nutrition of the Litopenaeus
vannamei in intensive shrimp ponds.
BIOFLOC AS AN ANTI-MICROBIAL AGENT
It was observed that the regular addition of carbon to the culture
is known to select for polyhydroxyalkanoate (PHA) accumulating
bacteria (Salehizadeh and Von Loosdrecht 2004) such as Alcaligenes
eutrophus, Azotobacter vinelandii, Pseudomonas oleovorans and others
that synthesise PHA granules.
THE DEVELOPMENT OF MICROBIAL FLOCS AFTER THE ADDITION OF CARBON SOURCE

Such granules are synthesised under conditions of nutrient stress, exhibiting bacteriostatic and/or bacteriocidal properties, depending
that is, when an essential nutrient like nitrogen is limited in the on the physiological status of the host and the physiochemical
presence of an excess carbon source. (Avnimelech 1999). These characteristics of the external environment (Ricke 2003).
PHAs are polymers of β-hydroxy short chain fatty acids and if These indicate that biofloc can be a novel strategy for disease
degraded in the gut, they could have antibacterial activity similar management on a long-term basis, in contrast to conventional
to short chain fatty acids (SCFAs) or organic acids. The breakdown approaches such as antibiotic, probiotic and prebiotic application,
of PHA inside the gastrointestinal tract can be carried out via especially since in many cases the efficacy of the latter two remains
enzymatic and chemical hydrolysis (Yu et al 2005). to be demonstrated.
Enzyme hydrolysis is generally carried out by PHA depolymerases Short chain fatty acids have been known for a long time to
produced by various micro-organisms such as Aspergillus fumigatus,
Pseudomonas fluorescens, Comamonas sp and others (Khanna and
Srivastava 2004). Chemical hydrolysis can be carried out by treating
the PHA with NaOH (Yu et al 2005). Pre-treatment of PHA
polymer with NaOH can significantly accelerate the digestibility
of PHA polymer. For example, an increase of up to 85 percent was
demonstrated in pigs (Forni et al 1999).
The working mechanism of SCFA with respect to their antibacterial
activity is not well understood. It could, however, be related to the
reduction of pH, as well as their ability to dissociate, or the pH of
the surrounding milieu (Ricke et al 2003). The antibacterial activity
increases with decreasing pH value. SCFA in undissociated form is
able to pass through the cell membrane of bacteria (Cherrington et
al 1991) and once internalised into the neutral pH of the bacterial
cytoplasm, it dissociates into anions and protons. These ions
influence the stability of functional macro-molecules and inhibit
the action of important enzymes. Bacteria need to pump out excess
protons to maintain the neutral pH of the cytoplasm. Removal of
protons requires consumption of cellular ATP and may deplete the
cellular energy and thus growth is inhibited.
Another possibility is that SCFA may interfere with membrane
structure and membrane protein in such a way that electron
transport is uncoupled and ATP production is diminished. Russel
(1992) speculated that anion accumulation is the primary reason
for the antimicrobial effect of organic acid. Moreover, less direct
antimicrobial activities have also been attributed to SCFA, which
include interference with nutrient transport, cytoplasmic membrane
damage resulting in leakage, disruption of outer membrane
permeability and hindrance in macromolecule synthesis (Ricke
2003).
Apart from inhibiting the growth of pathogenic bacteria by
lowering the pH of surrounding milieu, SCFA have also been
shown to specifically down-regulate virulence factor expression
and positively influence the gut health of animals (Teitelbaum
and Walker 2002). Moreover, these compounds are capable of

JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 9

RESEARCH FOCUS
THE POTENTIAL USE OF BIOFLOC AS AN ANTI-INFECTIVE STRATEGY IN AQUACULTURE
inhibit the growth of pathogenic bacteria and they also have been
used in commercial diets of terrestrial animals to control pathogens
such as Salmonella and E coli (Van Immerseel et al 2005) but their
use in aquafeed is very limited (on a commercial scale).
Recently, Defoirdt et al (2006) reported increased survival of
Artemia nauplii when fed formic, acetic, propionic, butyric and
valeric acid and challenged with a luminescence pathogenic Vibrio
campbellii strain. In another study, the same author (Defoirdt et
al 2007) reported that commercial polyhydroxy butyrate (PHB)
particles or PHB accumulating bacteria offered a preventive and
curative protection to Artemia against luminescent vibriosis.
They observed complete protection at a level of 1000mg/l of
commercial PHB or 107 cells/ml of PHB accumulating bacteria.
Although there are no reports of direct use of biofloc (PHA or
SCFA) as antimicrobial neutraceuticals in fish feed, many projects
have and are focussing on the characterisation and optimalisation
of PHA production by these flocs and analysing their bio-control
efficacy in different host-microbe systems.
CONCLUSION
In essence, biofloc technology could be useful in improving the
sustainability of fish/shellfish farming in both extensive and
modified extensive culture systems. There is clearly scope for
further improvement of this management strategy, not only
by optimising the quantity of carbohydrate addition at various
intensities of culture, but also by comparing the potential of
other carbohydrate sources. Additional research is required
with respect to feed composition planning and feeding rate
determination.
REFERENCES
Avnimelech Y 1999. C/N ratio as a control element in aquaculture
systems. Aquaculture 176. pp227-235
Burford MA, Thompson PJ, McIntosh PR, Bauman RH and Pearson
DC 2004. The contribution of flocculated material to shrimp,
Litopenaeus vannamei nutrition in a high-intensity, zero-exchange
system. Aquaculture 232. pp525-537
Cherrington CA, Hinton M, Pearson GR and Chopra I 1991. Short
chain organic acids at pH5 kills Escherichia coli and Salmonella
spp without causing membrane perturbation. Journal of Applied
Bacteriology 70. pp161-165
Defoirdt T, Halet D, Sorgeloos P, Bossier P and Verstraete W 2006.
Short-chain fatty acids protect gnotobiotic Artemia franciscana from
pathogenic Vibrio campbellii. Aquaculture 261. pp804-808
Defoirdt T, Halet D, Vervaeren H, Boon N, Wiele T, Sorgeloos P, Bossier
SUBSCRIBE ISSUE 21
NOW!
AC21.indd 1
Phone +64 9 533 4336 or visit www.skipper.co.nz
10 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
P and Verstraete W 2007. The bacterial storage compound poly-
b-hydroxybutyrate protects Artemia franciscana from pathogenic
Vibrio campbellii. Environmental Microbiology 9. pp445-452
Forni D, Bee G, Kreuzer M and Wenk C 1999. Novel biodegradable
plastics in sheep nutrition 2: Effect of NaOH pre-treatment of poly
(3-hydroxybutyrate-co-3-hydroxyvalerate) on in-vivo digestibility
and in-vitro disappearance (Rusitec). Journal of Animal Physiology
and Animal Nutrition 81. pp41-50
Hari B, Kurup BM, Varghese JT, Schrama JH and Verdegem MCJ 2004.
Effects of carbohydrate addition on production in extensive shrimp
culture system. Aquaculture 241. pp179-194
Khanna S and Srivastava AK 2004. Recent advances in microbial
polyhydroxyalkanoates. Process Biochemistry 40. pp607-619
Lancelot C and Billen G 1985. Carbon-nitrogen relationships in
nutrient metabolism of coastal marine ecosystems. In: Jannasch HW
and Williams JJL (eds). Advances in Aquatic Microbiology, vol. 3.
Academic Press, New York, USA. pp263-321
Mallasen M and Valenti WC 2006. Effect of nitrite on larval
development of the giant river prawn, Macrobrachium rosenbergii.
Aquaculture 261. pp1292-1298
Ricke SC 2003. Perspectives on the use of organic acids and short
chain fatty acids as antimicrobials. Poultry Science 82. pp632-639
Russel JB 1992. Another explanation for the toxicity of fermentation
acids at low pH: anion accumulation versus uncoupling. Journal of
Applied Bacteriology 73. pp363-370
Salehizadeh H and Van Loosdrecht MCM 2004. Production of
polyhydroxyalkanoates by mixed culture: recent trends and
biotechnological importance. Biotechnology Advances 22. pp261-
279
Teitelbaum JE and Walker WA 2002. Nutritional impact of pre and
pro-biotics as protective gastrointestinal organisms. Annual Review
on Nutrition 22. pp107-138
Van Immerseel F, Boyen F, Gantois I, Timbermont L, Bohez L, Pasmans
F, Haesebrouck F and Ducatelle R 2005. Supplementation of coated
butyric acid in the feed reduces colonisation and shedding of
Salmonella in poultry. Poultry Science 84. pp1851-1856
Yu J, Plackett D and Chen LXL 2005. Kinetics and mechanism of
the monomeric products from abiotic hydrolysis of poly [(R) -3-
hydroxybutyrate] under acidic and alkaline conditions. Polymer
Degradation and Stability 89. pp289-299
CONTACT Kartik Baruah. Email baruahkartik4@rediffmail.com ■
n JANUARY/FEBRUARY 2008
NATIVE
KOKOPU
$5.00
JANUARY/FEBRUARY 2008
IMMIGRATE
TO CANADA
SEA RESCUES
Land and sea
farmers share
RMA concerns
MOVE UP A NOTCH
Kaipara oyster
farm doubles Antarctica claims a cruise ship $10.50 (INCL GST)
national output $9.50
INCL. GST (NZ & AUST)
White Island tours – a successful achiever
ISSUE 61
Eel culture returns
to age old ways Chatham Island history revisited
WATERFRONT BUSINESS – THE LATEST IN NEWS AND VIEWS A SHOWCASE SELECTION OF THE LATEST WORKBOATS
9/12/07 9:49:18 PM
 NEW PUBLICATION
PEARL OYSTER HEALTH
MANAGEMENT MANUAL ON LINE
T
he FAO Fisheries and Aquaculture where pearl farms are located but also contributing
Department publication Pearl Oyster to maintaining environmental integrity. Pearl oyster
Health Management: a manual, is now farming can serve as environmental sentinels,
available online. The pearl oyster industry is a recognising the fact that pearl oysters thrive only in
growing multi-billion dollar sector of molluscan a pristine environment.
aquaculture. As pearl production relies on the This publication contains three parts. Part
health of the oyster, this manual, which serves as one consists of pearl oyster health – the current
a guide on the management of pearl oyster health, interest in it and an overview of the cultured
also includes an overview of the cultured marine marine pearl industry.
pearl industry, a general review of mortalities and Part two, pearl oyster health management,
disease problems to help reduce risks from disease, consists of seven sections:
and reports from Australia, the Cook Islands, • introduction
Japan, French Polynesia, the Philippines, China • general information on husbandry and
and other countries that have vast experience in handling, hatchery production, introduction
the health management of cultured pearl oysters. and transfers;
Pearl Oyster Health Management: a manual • disease diagnostic protocols dealing with
(FAO Fisheries Technical Paper No. 503. Rome, field collections of samples, gross external
FAO. 2007. pp120) was written by: examination, gross internal examination and laboratory protocols
• Melba G Bondad-Reantaso, Fishery Resources Officer • health zonation
(Aquaculture), Aquaculture Management and Conservation • disease outbreak protocols
Service Fisheries and Aquaculture Management Division, FAO • national strategies on aquatic animal health, and
Fisheries and Aquaculture Department Rome, Italy • references.
• Sharon E McGladdery, Aquatic Animal Health Division, Canadian Certain countries in the pearl oyster producing regions have
Food Inspection Agency Ottawa, Canada, and acquired a great deal of experience in health management of cultured
• Franck CJ Berthe, Animal Health and Welfare Panel, European species. Experiences from Australia, the Cook Islands, Japan, French
Food Safety Authority Parma, Italy. Polynesia, the Philippines, China, the Persian Gulf and the Red Sea
The pearl oyster industry is a growing multi-billion dollar sector are included in part three, which also contains a general review of
of mollusc aquaculture. Pearl farming occurs throughout Australasia, pearl oyster mortalities and disease problems.
the Middle East and South America. Few species of molluscs possess The manual can be downloaded in full or in part via the FAO Fisheries
the ability to produce pearls of gem quality. The South Sea pearl and Aquaculture Department website: http://www.fao.org/fishery ■
oyster is one of them.
Pearl production in the wild is an unpredictable and uncontrolled
event which human intervention, through pearl culture, has
progressively overcome by improving culture practices. Farming
mother-of-pearls shares commonalties with edible mollusc
aquaculture. However, the end product, pearl production, is unique
to this sector. In aquatic production, health issues are of utmost
importance; pearl production is based entirely upon health.
The pearl itself is a product of the oyster's immune defences as a
response to soft-tissue irritation. Exploited stocks receive frequent
handling stresses that often predispose farmed animals to infection
and diseases. The importance of health management for pearl
oysters is therefore paramount.
Today, most disease problems are caused by opportunistic
pathogens taking advantage of oysters weakened by the stress
of handling, including pearl surgery and sub-optimal growing
conditions. Except for the mass mortalities experienced in Japan, the
pearl oyster industry has not yet faced the types of epizootics that
have impacted mollusc culture elsewhere in the world. Development
of the industry will inevitably lead to increased risk of disease
introduction, spread or emergence. Health management is the
C O N S U LTA N C Y • T R A I N I N G • P U B L I S H I N G
critical defence line against such an unwanted future.
This technical paper has two objectives. The first is to review
pearl oyster mortalities and disease problems in order to help design PATTERSON PEDDIE CONSULTING LTD.
programmes aimed at reducing the risks from diseases. The other E-mail: info@pattersonpeddie.com
objective is to provide technical guidance to pearl oyster farmers and the Tel: +44 (0) 2893 351379 (Office)
industry on managing pearl oyster health so that sector development will w w w. p a t t e r s o n p e d d i e. c o m
be sustainable, not only in providing huge employment to communities

JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 11

RESEARCH FOCUS

THE BC PACIFIC SALMON

FORUM, NANAIMO, CANADA
Collaborative Research Initiative in British Columbia Deepens Understanding of the
Broughton Archipelago and Throws Light on Salmon and Sea Lice Issues

I
n 2007 the BC Pacific Salmon Forum, an independent citizen
body using science and stakeholder dialogue to advance sustainable
governance of BC Pacific salmon, launched a two-year collaborative
research programme to improve our understanding of the Broughton
Archipelago ecosystem.
The programme engages over 30 scientists and many others, with
a variety of disciplines and perspectives, in some 15 research projects.
THE BROUGHTON ARCHIPELAGO
They include defining oceanographic dynamics, monitoring salmon,
stickleback and sea lice population dynamics, investigating the impacts
of lice on salmon, and performing surveys of wild fish health. A number
of key findings have been identified from individual projects in 2007,
and although the data is preliminary and study will continue in 2008,
some key points of interest arose.
The following is an overview of the preliminary findings from the
2007 research period. (See end of article for contact details.)

ECOSYSTEM DYNAMICS
Oceanographic factors play a significant role in the dynamics of any
ecosystem, and in particular, in the distribution of organisms (both
fish and potential pathogens such as sea lice) within that system.
The Broughton Archipelago is a complicated system and to better
understand it, and to provide insights into the distributions of fish and
lice, several research groups have gathered oceanographic data.
The data indicates a considerable degree of wind-driven surface water
circulation that may result in surface transport in directions counter to
tidal currents in the Broughton Archipelago. Although currents near
the surface move predominantly in a seaward direction, winds can
drive surface water (and potentially any particulates, including sea lice,
contained in these surface waters) up inlets contrary to the current
flow.
Oceanographic fluctuations have significant impacts on organisms
living within an ecosystem. A delicate balance exists between a fish,
the environment it lives in and potential pathogens. It is an accepted
fact that any change (environmental, nutritional or physiological) that
pushes an organism beyond its ability to cope results in stress, which can
upset homeostasis and lead to increased pathogen susceptibility.
A comparison of data between 2003 and 2006 demonstrated that
surface water salinity and surface water temperatures were greater in
2004 compared to other years, and this corresponded with an increased
abundance of sea lice. Surface water temperatures increased (from 8.5˚C
to 12.5˚C) and surface salinity decreased (from 26ppt to 19ppt) between
April and June 2007.
SEA LICE STUDIES
Plankton tows were performed to identify the distribution patterns
of planktonic lice, both near to and distant from salmon farms. The
majority of early samples (February and March) contained no lice, but
MAP OF THE
BROUGHTON ARCHIPELAGO
although larvae were relatively rare, some patterns did emerge. Caligus
copepodids were found in higher abundance in side inlets and low
salinity areas, while Lepeophtheirus copepodids were most abundant
near active farms.
Plankton tows in April were hampered by dense plankton blooms
that congested nets and precluded further sampling. In 2008, new
equipment will allow the examination of vertical distributions of sea lice
larvae in the water column. Their salinity and temperature preferences
and their diel (ie daily) migration patterns will also be investigated.
Several projects involved collecting wild juvenile pink and chum
salmon and evaluating their infestation rates. Although there was
some degree of variation between research groups with respect to the
prevalence and intensity of lice observed on wild pink salmon captured
in 2007, lice levels were low early in the season (March and April) and
increased over time (April to June).
Overall, the prevalence of lice on juvenile pink salmon in 2007
was described as being low, relative to previous years. In one study
approximately 80 percent of all juvenile pink and chum salmon
captured had no lice (ie the prevalence was approximately 20 percent).
This represented the overall infection rate (prevalence) observed for

12 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008

ABOVE: SALMON SMOLTS IN A SEA CAGE
RIGHT: SAMPLING
all pink and chum salmon caught
between March and June, including
fish captured close to and distant from
salmon farms. Juvenile chum salmon
appeared to be more heavily infected
than other species in all the years
examined.
Concerns have been raised in the
past that different identification
methodologies may preclude the
ability to compare data; a comparison
of sampling methods for fish and
lice identification and quantification
was performed by two different
investigation teams that resulted
in almost complete agreement in
findings.
Two further studies found similarly low lice prevalence (<25 percent
to 31 percent), and a fourth study that collected juvenile pink and chum
salmon for survival studies described an abundance of lice on wild-
caught fish as generally being less than one louse per fish, and chalimus
and copepodids as declining, with motiles rarely exceeding an overall
ratio of 0.4 lice/fish over 20 days.
Sampling of lice prevalence on wild fish was carried out by all research
teams both near to and distant from salmon farms, but a statistical
analysis of transmission estimates has not yet been completed.
Lice levels in the Broughton Archipelago were found to differ
compared with another geographical region on the mid-BC coast.
Although this area encompasses different environmental conditions
from those in the archipelago, the prevalence of sea lice on juvenile
pink and chum salmon in the Bella Bella region averaged 4.2 percent,
with lice slightly more prevalent on chum than pink salmon in this
area. Statistical analysis of the differences in lice prevalence has yet to
be undertaken.
IMPACTS OF SEA LICE ON JUVENILE SALMON
There has been significant debate on the impacts of sea lice on juvenile
salmon and there is not yet complete agreement on the sufficiency of
current data to allow firm conclusions.
It is common in nature to find fish with parasites, yet these organisms
must maintain a fine balance lest they overwhelm their host. Similarly,
the host, if in a sufficient state of fitness and health, can maintain
homeostasis in the face of pressures exerted by a reasonable number
of parasites. What defines “balance”, “fitness”, “health” and “reasonable”
with respect to these species is not yet completely known, as these
specific research questions are relatively recent. Part of the research
SAMPLING JUVENILES
EXAMINING JUVENILES
being undertaken by the BC Pacific Salmon Forum is designed to
address these questions.
A health survey conducted in the region reported that both the
weight and the length of juvenile pink salmon increased between April
and June. The condition factor (one measure of the physical condition
of fish that describes the relationship between body weight and body
length) was significantly associated with the sampling month, while the
presence or absence of sea lice and sea lice density (lice/g of fish) was not
significantly associated with the condition factor in any general linear
model tested.
Skin lesions were found to be associated with the presence of sea lice,
as were lower liver glycogen stores (energetic reserves). No other health
parameters (ie the condition factor, histopathological findings, viral or
bacterial infections) were apparently associated with lice.
Histological studies were performed on saltwater entry juveniles
and, based on the findings, fish first entering the marine environment
appeared healthy. A portion of those fish sampled one month later
displayed liver lesions consistent with toxicants. There was a positive
correlation between some liver lesions and a myxosporean parasite (25
percent of fish were infected with the myxosporean) but in no cases were
liver lesions or myxosporean infections correlated to lice infestation.
Additional sampling is scheduled in 2008 to provide a stronger statistical
basis for analysing these relationships.
Wild pink salmon have been successfully transferred to and raised in
a laboratory setting displaying growth patterns similar to those observed
in the wild. Two studies that collected and held naturally infected wild
fish found that lice disappeared over approximately two weeks.
Repeatable methods of measuring swimming performance were
developed and, when fish with one louse attached were compared
JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 13
RESEARCH FOCUS
THE BC PACIFIC SALMON FORUM, NANAIMO, CANADA
ABOVE LEFT: LICE COUNTS
ABOVE CENTRE:
JUVENILE SALMON UNDERGO A FITNESS TRIAL
ABOVE RIGHT: SAMPLING FOR JUVENILE SALMON
RIGHT: A SEA LOUSE IS VISIBLE ON THIS INDIVIDUAL
FAR RIGHT: JUVENILE SALMON
with fish having no lice, there were no significant differences in their
maximum swimming performance. Louse stage (at one louse per
fish) had no apparent effect on maximum swimming performance.
Copepodids artificially raised from egg strings were successfully
used to infect juvenile pink salmon in the laboratory, and while the
duration of survival of juvenile salmon did decrease with increasing
lice loads, this effect was not statistically supported unless lice loads
were 10 times greater than levels reported in nature. Interestingly,
mortalities of pink salmon spiked when the lice entered a moulting
stage of their development.
An examination of the impact of sea lice on the schooling behaviour
of juvenile pink salmon in tanks in the field was carried out, and
preliminary findings suggest that predation may occur selectively
on lice-infected fish. This data has not yet been fully analysed, and
modelling is necessary to substantiate the results and evaluate the
implications more thoroughly.
Salmon populations can and do fluctuate for a great number of
reasons, many of them environmental. Variations in the freshwater
environment can impact early rearing stages of fry, environmental
conditions on marine entry can impact what is known to be a
physiologically challenging period, oceanic productivity can impact
prey availability and parasite populations, and the high seas are a black
box into which we have yet to accurately peer.
The overall catch of juvenile salmon in the Broughton Archipelago,
for sampling purposes in 2007, was described as being the lowest over
the previous five-year period of monitoring (despite an increase in the
number of sampling sites) and there was greater variation in the size of
fish than previously observed.
Freshwater counts from a variety of systems and the migratory
patterns of adult and juvenile salmon were tracked through the
installation of video equipment, an acoustic counter, aerial surveys and
fry traps.
The 2007 pink adult salmon returns (escapement), as a whole,
in the mainland inlets of the Broughton Archipelago system are
described as being similar or slightly improved, relative to the
brood return in 2005. The out-migration from the Glendale River
spawning channel was estimated at 7.7 million pink fry and 34,000
chum salmon fry during 2007, but fry appear to have migrated out
later in 2007 than in past years.
The three spine stickleback fish has been of interest for several years,
following numerous reports of them being heavily infested with lice.
14 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
In some cases sticklebacks have been described as being, “feathery with
lice”. There is no evidence to date that lice are capable of carrying out
their entire life cycle on the stickleback, and the stickleback’s heavy
armour is thought to protect it.
In 2007 sticklebacks were captured in large numbers during juvenile
salmon fry sampling and numbers were high relative to previous
sampling years. In many cases sticklebacks were reportedly more heavily
infested by lice than juvenile pink and chum salmon captured in the
same sets.
Interestingly, in a captive study, sticklebacks were observed
cropping egg strings from gravid females on juvenile salmon and
actively removing adult lice in some instances. It is not known what,
if any, role sticklebacks play in the transmission dynamics of lice in
the Broughton.
CONCLUSION
The 2007 research period has ended and much of the data is still
being analysed. It is difficult, if not impossible, to make well-founded
statements about the dynamics of organisms in a region without
accurate assessments of populations and the environment over time.
For this reason, the majority of the projects being funded by the
Forum are being repeated for a second year to provide more data. The
2008 research programme is now underway, and new data emerging
over the coming year will add to our expanding knowledge of this
complex system.
The Broughton Archipelago is very intricate, and its currents,
winds, geography and ecology are very complicated. The Forum
has stated that, “Since it is clear we are dealing with a dynamic
ecosystem that includes many factors, not simply sea lice, funding is
being allocated in 2008 to engage a range of researchers to develop
an analytical framework that will incorporate all possible ecosystem
factors in order to interpret the data that is emerging from this
research programme.”
This analysis would be a first attempt to undertake a whole system
evaluation of many of the key environmental factors that affect salmon
in the Broughton Archipelago.
While research will not unravel all of the ecological mysteries of lice
and salmon in the archipelago, perhaps some of the more pressing
questions can be, at least in part, addressed through this scientific
process.
See www.pacificsalmonforum.ca/research/index.php ■

NEWS ROUNDUP
BY SUZI FRASER DOMINY
This news roundup appears courtesy
of the Aquafeed website. See www.aquafeed.com
SALMON GET TRAINING TIPS
FROM FOOTBALL
What do juvenile salmon and the Spanish football team Barcelona
have in common? Both use intensive training to improve their
form. The head coach is a Nofima scientist and the training is
healthy for fish, too.
In order to make salmon more robust for transfer to sea water, an
interdisciplinary research group involving Nofima, the Norwegian
Institute for Water Research (NIVA), the Norwegian School of
Veterinary Science, NTNU, Thelma AS and Aakvik Settefisk AS
decided to strengthen the salmon’s heart capacity and health
through intensive training. Benefits to humans through training
include strengthening of the heart, muscles, skeleton and immune
system and reduction of stress.
“We were concerned that the fish would develop lifestyle diseases,”
says senior scientist Harald Takle of Nofima Marin (formerly
Akvaforsk), who headed the research group. The scientists have
found hearts in farmed salmon that differ from the heart form of
wild salmon, so the thought that the fish needed training wasn’t far
off the mark. It was just a matter of starting to train 50g salmon.
The scientists increased the tank water velocity to get the fish to
swim faster. One of the challenges was to get tiny heart rate monitors
made. These were inserted into the fish’s stomach with sensors
attached to the heart.
The fish in the trial were divided into three groups:
• a control group that lived the normal life of juvenile salmon
• one that received increased tank water velocity around the clock,
and
• one also received continual “jogging”, a daily spell of
high-intensity training.
The findings showed that the fish that trained grew
considerably quicker, but that it did not influence feed
The new knowledge about juvenile
utilisation. salmon could lead to salmon farmers
“We are now conducting tests at the VESO Vikan
fish farm to see if the fish with headbands and heart putting more jets in fish tanks and
rate monitors have greater powers of resistance to a
deadly virus. The findings are extremely promising. regulating the tank water velocity
Moreover, the fish tackle stress better when they are
in better form. Less stress means the fish have greater
energy reserves to tackle the challenges of everyday life,” says Takle.
The new knowledge about juvenile salmon could lead to salmon
farmers putting more jets in fish tanks and regulating the tank water
velocity. “In the long term, we believe that this can make the fish even
more robust. It’s just like with us humans, healthier fish thrive better,
and this will in turn increase profitability for the salmon farmers,”
Takle said.
AQUA BOUNTY SCALES BACK COMMITMENT
SemBioSys Genetics Inc., a biotechnology company developing a
portfolio of therapeutic proteins for metabolic and cardiovascular
diseases, says its joint development partner on the ImmunoSphere
shrimp feed additive programme, Aqua Bounty Technologies Inc,
intends to scale back marketing and registration efforts on its first-
AQUAFEED
generation shrimp feed additive, IMS. SemBioSys is developing
a second-generation shrimp feed additive product in safflower,
called ImmunoSphere, which it says could significantly reduce the
manufacturing costs of the feed additive.
“As a result of the decision, SemBioSys Genetics intended to
evaluate the impact it had on ImmunoSphere,” said the president
and chief executive, Andrew Baum. ImmunoSphereTM offered
a significant reduction to the cost of producing immune-
ostimulatory feed additives, Baum said. “We intend to evaluate
all the available options prior to committing to the next steps
with this product, which targets a large potential market that has
a significant unmet need.”
The global shrimp industry represents a US$18 billion market.
One of the major challenges facing commercial shrimp producers
is the outbreak of diseases, which cost the shrimp aquaculture
industry more than US$3 billion per year. Immuno-stimulatory feed
additives, like ImmunoSphere, are designed to provide prophylactic
protection to shrimp production.
PROSOL CREATES NEW PRODUCTS
BASED ON NUCLEOTIDES
The Italian company Prosol, which is a private producer of natural
nucleotides for dietary food applications, has launched a new range
of nucleotide-based products for aquaculture. Based on well-known
independent scientific literature, these products provide a specific
source of Free 5' nucleotides (F5N). It has been proved that only
nucleotides in free form enhance the immune system’s response to
stress factors.
Nucleo 5 Prime, the latest in the Prosol range, is developed from
the company's deep knowledge in nucleotide extraction technology.
In response to feed producers’ requirements, Nucleo 5 Prime is said
to offer:
• expertise in production
• a constant ratio among free 5' nucleotides
• a precise amount of free Single nucleotides
• determination of free 5' nucleotides in each batch
• GMP+ certification, and
• optimal ratio quality/price.
Nucleo 5 Prime is obtained in its free form by enzymatic hydrolysis
of ribonucleic acid in yeast. Nucleo 5 Prime is then spray dried and
provided in two different concentrations: six percent F5N and 40
percent F5N content. ■
JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 15
new INTERNATIONAL
Skretting tests marine zooplankton
against lice
Skretting has entered into a contract with Calanus AS to
develop and document a product based on the marine
zooplankton Calanus finmarchicus to counteract sea lice
infestations in salmon and trout. Commercial-scale field trials
by selected farmers in Chile and Norway are now starting.
The pelagic stages of Calanus finmarchicus and the infective
stage of salmon lice are derived from a common ancestor
and have many similarities. Therefore they may share what
researchers call immunological structures. The trials will
test whether it is possible to achieve higher protection from
salmon lice by stimulating the immune system of the salmon
with this new zooplankton product.
It is known that the immune system of mammals can be
stimulated to improve protection against external parasites. It
has also been documented that the immune system of salmon
can be improved to repel lice attacks.
“The development of new, alternative salmon lice
treatments is important. Although the outcome of this
project is unknown, we believe the idea is too good not to try
it out,” said David Knudsen, Skretting’s international product
manager, raw materials.
Sea lice infection is a major problem for the fish-farming
industry in Chile, Canada, the United Kingdom and Norway.
If the test results are sufficiently positive, the product will be
launched for commercial use.
“We are very pleased with the agreement and that Skretting
is willing to invest significant resources to test our product on
a commercial scale in an attempt to help fish farmers solve a
serious problem,” said the managing director of Calanus AS,
Gunnar Rørstad.
Disease affects Marine Harvest results
Marine Harvest says the on-going disease situation in Chile
continues to impact the group, giving a negative operational
EBIT of 65 million kroner in the first quarter. The situation
in Chile remained a challenge, but Marine Harvest said it was
still confident of a positive long-term outcome.
The largest operation, Marine Harvest Norway, continued
to deliver good figures in the first quarter of 2008, with higher
harvest volume and lower cost of harvested fish than the same
period last year. VAP Europe also continued to show positive
development with an increased EBIT.
“We have taken a number of operational actions to
contribute to short and long-term improvements in Chile, but
it takes time to achieve the envisaged improvements,” said the
chief executive, Åse Aulie Michelet. “The number of new ISA
sites identified has been reducing in the last months, however
the overall situation remains uncertain. Still, we are optimistic
about the long-term outcome for Chile as an attractive and
profitable production region.”
Michelet said the company had taken strong action to
reduce costs in the quarter, including closing a processing
plant and making 900 staff redundant.
Marine Harvest Norway had shown good operational
improvement compared with last year. The reduction in
operational EBIT was to a large extent due to lower market
prices and currency effects.
“We are further pleased with the results in VAP Europe that
improved its EBIT, despite a lower revenue compared to the
same period last year,” said Michelet.
Marine Harvest generated operating revenues of 3080 million
kroner in the first quarter 2008 compared to 3795 million
kroner for the same period last year. The reduction in revenues
16 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
of 715 million kroner is mainly a result of lower prices in all
regions. The reduction in revenues in Chile alone amounted to
594 million kroner, although a large part of this reduction was
also due to lower volumes and smaller fish harvested.
EBITDA before fair value adjustment of the biomass and
restructuring was 107 million kroner in the quarter, down from
740 million kroner in first quarter 2007. Operational EBIT was
-65 million kroner in the first quarter of 2008 compared to
positive 552 million kroner in the first quarter of 2007.
The total harvest volumes in the first quarter were similar to
the levels of the comparable period last year, as the reduction
in Chile due to disease was compensated by a volume increase
in Norway.
At the end of the first quarter 2008, Marine Harvest had a
net interest-bearing debt of 6314 million kroner compared to
6725 million kroner in the first quarter 2007 and 6744 million
kroner at the end of 2007. The equity ratio was 53.8 percent
compared to 54.7 percent in the first quarter 2007.
Marine Harvest is divided into five business areas: Marine
Harvest Norway, Marine Harvest Chile, Marine Harvest Canada,
Marine Harvest Scotland and Marine Harvest VAP Europe
(value added products). It also has operating units in Ireland, the
Faroe Islands and Japan, and halibut farming in Norway.
Marine Harvest Norway had operating revenues of
1228 million kroner for the first quarter. Operational EBIT
was 93 million kroner. The harvest volume in the period was
a record high for a first quarter at 40,353 tonnes (HOG),
contributing to an EBIT/kilogram (HOG) of 3.16 kroner.
EBIT levels were lower than the 2007 first quarter due to
reduced market prices and currency impacts.
Marine Harvest Chile had operating revenues of 568
million kroner for the first quarter. Operational EBIT was
-155 million kroner. The sold volume in the period was
23,883 tonnes (HOG), giving a negative EBIT margin of
EBIT/kilogram 6.48. The negative result was impacted by
lower market prices, smaller fish size, lower volumes and
inventory write-downs.
Marine Harvest Canada had operating revenues of 318
million kroner for the first quarter. Operational EBIT was 19
million kroner. The harvest volume in the period was 11,348
tonnes (HOG), contributing to an EBIT/kilogram of 1.64
kroner. The biological situation in Canada remains good.
Difficult circumstances with lower prices and the falling US
dollar compared with the Canadian dollar had a negative
effect on profitability in the first quarter.
Marine Harvest Scotland had operating revenues of 184
million kroner for the first quarter, and operational EBIT was
-35 million kroner. The harvest volume in the period was 6275
tonnes (HOG), resulting in an EBIT/kilogram negative margin
of 5.54 kroner. Scotland’s underlying performance showed
improvement, but performance figures were influenced by
an incident that led to a product recall. The result included
provision for 34 million kroner to cover the costs of the recall.
Marine Harvest VAP Europe had operating revenues of
839 million kroner for the first quarter. Operational EBIT
was 29 million kroner, giving an EBIT margin of 3.4 percent
compared with 1.4 percent in the corresponding period
last year. The increase in EBIT margin despite the reduced
revenues in the quarter was satisfactory, the company said.
Marine Harvest expected to harvest 319,000 tonnes of
salmonids in 2008, an increase of 11,000 tonnes compared
with the previous year, of which 73,000 tonnes would be
harvested in the second quarter.
“The situation in Chile remains uncertain. The frequency
of new ISA outbreaks has been reduced over the last months.
However, 2008 will be a challenging year for Marine Harvest
Chile. The restructuring of our operations in Chile has the
highest priority,” Michelet said.
Feed costs were a challenge for salmon farming as well as
for other protein farming industries, and Marine Harvest had
for the first time negotiated global agreements with its feed
supplier. “This will be important for the attention we will pay
to the use and composition of feed in the future.”
Marine Harvest anticipated a breach of one of the covenants
in the loan agreement by the end of the first quarter of 2008.
However, the company had obtained a waiver from the bank
syndicate of this financial covenant in relation to the first
quarter of 2008.
“We are in a constructive dialogue with the banks to get the
current covenants amended.” The global supply of Atlantic
salmon grew by five percent compared to the first quarter last
year, significantly below recent growth rates.
“As a result of the situation in Chile we reiterate a global
supply growth of one or two percent for the year and
improved market balance in the main markets.”
The board expected improvement in the EBIT for the
second quarter compared to the first quarter, reflecting higher
prices and slightly lower costs. “However, several measures
have to be taken to meet the long-term financial targets for
this business.”
New team at Intervet/Schering-Plough
Schering-Plough has appointed the executive team members
of the company’s global animal health unit: Intervet/Schering-
Plough Animal Health. These appointments define the major
steps in the design process of the animal health unit.
The executive team is chaired by Ruurd Stolp, senior vice-
president and president animal health. Dr Stolp reports to
the chairman and chief executive of Schering-Plough, Fred
Hassan. The executive team members are
• Raul Kohan, senior vice-president corporate excellence, and
deputy head animal health
• Hugo Wahnish, vice-president global AH regions
• René Aerts, vice-president animal health, research and
development biologicals
• KJ Varma, vice-president AH, research and development
pharmaceuticals
• Jochen Bader, vice-president global finance, animal health
• Malte Greune, vice-president global supply chain animal
health
• Gráinne Higgins, vice-president HR, animal health
• Mark van Heumen, law lead global animal health, and
• Andreas König, quality head, global quality operations,
animal health.
The new combination of Intervet/Schering-Plough Animal
Health strengthens the animal health portfolio in several areas
and brings together complementary lines of pharmaceuticals,
biologicals and innovative services, the company says.
Schering-Plough completed its acquisition of Intervet last
November. The American company purchased the animal
health division from the Dutch chemical giant Akzo Nobel
for €11 billion.
UNITED STATES
PerOs granted licence
for oral vaccine formulations
PerOs has been granted an establishment licence to produce
bacterins as well as an autogenous bacterin licence utilising its
patented oral delivery formulation methods (Oralject).
The Centre for Veterinary Biologics at the United States
Department of Agriculture granted the licences in April.
“The approval of our manufacturing facility and our oral
vaccine formulation platform in the United States marks
a significant milestone for our company,” said the chief
scientific officer, Grant Vandenberg, who invented Oralject.
“We look forward to offering these capabilities to the livestock
producers and opening new possibilities to the growing
aquaculture market, as well as collaborating with traditional
animal health companies to expand the oral enablement of
the industry as a whole.”
PerOs’ oral delivery platform focuses on the use of natural
products to deliver safe therapeutic and prophylactic compounds.
Its formulations are designed to enable the delivery of biological
compounds such as vaccines, peptides and probiotics, among
others, while eliminating the stress associated with handling,
injection and other current delivery practices.
The company says the new technology platform will
enhance the value of vaccines by offering a labour-saving,
practical oral delivery approach for the livestock industry and
open up new possibilities to the rapidly growing aquaculture
market. Oral vaccination provides animal producers with an
innovative tool for disease prevention through vaccination,
rather than relying on disease treatment using antibiotics.
The PerOs facility, said to be the first of its kind, was
made possible by the contribution of Benchmark Biolabs’
development team. Benchmark managed the regulatory
interface and established the QA and QC standards, while also
designing, constructing and now operating the PerOs plant.
The chairman of PerOs, Ian Kott, said the work of the
Benchmark team was “perfect. Using their BEAM process
flow, they led our technology assets in record time toward
regulatory approval.”
Contact Jean-Simon Venne, PerOs Inc. Phone (514) 516
3003, email jsvenne@perosbio.com, or Tom Overbay, DVM,
Benchmark Biolabs, Inc. Phone (402) 475 8104 or email
toverbay@benchmarkbiolabs.com
Ohio extends emergency order
Ohio’s Agriculture Director, Robert Boggs, extended an
emergency order on May 16 that prohibits the intra-
state transportation, sale or distribution of 28 fish species
susceptible to viral haemorrhagic septicemia, or VHS, out of
the affected region in northern Ohio.
The Ohio Department of Agriculture is responsible for news
protecting animals from potentially devastating diseases. “This
emergency order not only protects Ohio’s fish population but
also others in points beyond the state’s borders,” Boggs said.
VHS is a fish disease that must be reported to the
department under state law. It was introduced into the wild
fish population by an invasive species. It is not harmful to
humans or other animals. Tests by the ODA’s Animal Disease
Diagnostic Laboratory has revealed that VHS is not present
inland in Ohio. The department will continue to test and
monitor for the disease.
Ohio’s ban prohibits the intra-state distribution of VHS-
susceptible fish or eggs out of the area in Ohio north of US
Highway 6 from the Indiana border to the intersection of H6
and Interstate 90 near Fremont, continuing on I-90 to the
Pennsylvania border. This includes the Sandusky River south
of H6 to the Ballville Dam.
Twenty-seven fish species are known to be susceptible
to VHS. The prohibition is in effect in Ohio until the US
Department of Agriculture’s Division of Animal and Plant
Health Inspection Service releases its embargo and prohibition
of the movement of live VHS-susceptible fish.
The prohibition does not apply to live fish or eggs removed
directly from production facilities that have tested negative
for VHS. It also excludes live fish or eggs that are being
JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 17
new transported for use by research scientists in closed research
facilities with diagnostic laboratories.
See www.ohioagriculture.gov
CHILE
New general manager
for Pharmaq Chile
Dr Bernd Wrege has been appointed general manager of
Pharmaq Chile Ltda, situated in Puerto Montt. He will
continue his responsibilities as technical director. Dr Wrege
has held key positions within the company since 2004, and has
extensive experience within the aquaculture industry in Chile.
Pharmaq Chile Ltda is as a subsidiary of the Norwegian
company Pharmaq AS, and started its activities in Chile in
2001. The company says it has experienced a strong growth in
the Chilean market and occupies the number one position in
vaccines and treatment against Caligus.
The aquaculture industry in Chile is facing several challenges
related to diseases, and the company says it is committed to
helping the industry by introducing new products and
customer services.
Pharmaq introduces vaccine
in a new product line
Pharmaq Chile Ltda has been granted a licence by Servicio
Agricola y Ganadero de Chile (SAG) for the new vaccine
Alpha Ject® micro 3. This is a three-component vaccine for
salmon in a micro-dose (0.05ml/dose) to protect against
infectious pancreatic necrosis, vibriosis caused by Vibrio
ordalii, and SRS caused by Piscirickettsia salmonis.
“With all the efforts and resources put into this project
we are confident that we have developed an effective and
safe vaccine for our customers,” said the company’s general
manager in Chile, Dr Bernd Wrege.
Alpha Ject® micro 3 represents the first micro-dose SRS
vaccine delivered from Pharmaq. “The documentation of
this new-generation micro-dose product from Pharmaq
is comprehensive. We had to develop new technologies to
ensure the right concentrations of antigens for optimum
protection of this product.”
As well as ensuring the efficacy of fish vaccines from
Pharmaq, all the products go through laboratory and field
tests to establish safety documentation as well as safeguard the
maximum growth of the fish.
The introduction of the new vaccine follows the company’s
launch one year ago of the first SRS vaccine, Alpha Ject® 4-1.
Contact Bernd Wrege, general manager Pharmaq Chile Ltda.
Phone (+56) 65 483091, or email bernd.wrege@pharmaq.cl
UNITED KINGDOM
Isolation of spring viraemia of carp virus
Recent routine import checks have identified spring viraemia
of carp, or SVC virus in goldfish imported from a supplier in
Hong Kong, the Tung Hoi Aquarium Company.
SVC has no implications for human health. It is nonetheless,
a serious viral disease affecting common and ornamental carp,
as well as tench, roach, rudd, goldfish, pike and wels catfish.
The virus was found in a random sample taken directly from
imported boxes of fish. The Fish Health Inspectorate at Cefas
has placed movement controls on, and taken samples from, all
sites receiving fish from the SVC-positive consignment. The
FHI has also provided advice to other businesses that may
have recently imported fish from the same source to increase
awareness of any potential SVC risk.
The authorities in the exporting country have been informed
of the isolation of SVC and are conducting investigations to
18 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
identify the source of the infection. Importation of fish from
the affected source will be prohibited pending the conclusion
of these investigations.
The clinical signs of SVC can include darkening of the skin,
swollen eyes, abdominal swelling, pale gills, trailing faecal casts
and protrusion of the anus. Infected fish may be lethargic and
show areas of bleeding in the gills and skin.
Anyone noting deaths in carp or any other species susceptible
to SVC, with signs of disease similar to those above, should
immediately contact the Cefas Fish Health Inspectorate at the
Weymouth laboratory on 01305 206673/4.
Anyone who imports, keeps or retails carp or other
susceptible fish should take strict precautions to prevent the
spread of SVC and follow the advice set out in Defra’s booklet
Keep Fish Disease Out, available from the Cefas Fish Health
Inspectorate or through www.efishbusiness.com
Salmon industry to invest £400k in
research and development
Scottish salmon farmers are to benefit from a £400,000
investment in research and development projects over the
next three years to help secure the long-term socio-economic
and environmental sustainability of the industry.
“This project will be highly significant in developing new
farming practices in the Scottish industry,” said the chief
executive of the Scottish Salmon Producers’ Organisation,
Sid Patten. “It is hoped that the information generated will
enhance sustainability, while improving both environmental
performance and reducing the cost of production.
“To be successful, it must report good quality, robust
scientific evidence. By working closely with stakeholders, the
initiatives address many of the current challenges and they will
help the industry to achieve its full potential,” Patten said.
A range of initiatives has been identified, including a
scientific environmental modelling project and the adoption
of new technology and husbandry practices on farms. An
enhanced monitoring programme has been commissioned to
develop the accuracy of predicting environmental performance
of marine farms. A second initiative is to look at economic
modelling of production to determine optimum sustainable
farming conditions.
Projects addressing alternative sea lice management
strategies, the application of new technology in freshwater
farms and further development of acoustic devices to deter
predators are under consideration.
The initiative is being managed and partly funded by
the producers’ organisation under the auspices of the new
Demonstration Project, with financial and other support from
the Scottish government, salmon farmers, industry suppliers,
Highlands and Islands Enterprise, The Crown Estate, Scottish
Aquaculture Research Forum and regulators.
Scientists gather for workshop
Marine scientists from around the world took part in a
special workshop in Weymouth on infections and diseases
in marine life during May. The 27 delegates took part in a
histopathology workshop at the Centre for Environment,
Fisheries and Aquaculture Science Laboratory.
The three-day workshop featured presentations from
experts, laboratory tours and breakout sessions, and placed
significant emphasis on “hands-on” practical training and the
diagnosis of diseases and pathological changes. Participants
studied tissue samples through microscopes and examined
tissue sections under high-magnifying monitors.
Species examined included salmonids, cyprinids, molluscs
and crustacea. One of the organisers, pathologist John
new Bignell, said that scientists came from as far afield as Tasmania
and South Africa to learn about histopathology.
Gettinby honoured with award
Professor George Gettinby from the University of Strathclyde
in Scotland was honoured with
the Intervet Dieter Lütticken
Award at World Aquaculture
2008, held in Busan, Korea from
May 19 to 23. The award, which
carried a prize of €20,000, was
for his innovative statistical
and epidemiological research
methods in sea lice treatments in
salmon farming. The jury found
his research methods were an
PROFESSOR
excellent example of commitment
GEORGE
to the advancement of reducing, GETTINBY
refining and replacing the use
of animals in research, development and production of
veterinary medicines.
Professor Gettinby’s research focused on applying statistical
and epidemiological methods in sea lice treatments in salmon
farming in order to minimise the number of animals required in
test groups. The research also provides an alternative to assessing
the effectiveness of veterinary medicines in fish, which supports
the replacement principle. This alternative is the adoption
of mathematical models, which can simulate the effects of
treatments on lice populations.
Dr Dieter Lütticken, who presented the award, said there
had been a long and intimate relationship between life sciences
and statistics. “Professor Gettinby’s innovative statistical and
epidemiological research methods are an excellent example of
refining and reducing the use of animals in sea lice treatments
in salmon farming.”
The jury panel praised Professor Gettingby’s innovating
mathematical model approach for investigating the optimal
use of treatments that involve the minimal use of animals. ”In
addition, these methods also minimise the use of veterinary
medicines in the environment.”
Professor Gettinby said he was honoured to receive the
Dieter Lütticken Award. “It is pleasing to know that the
award has come as a result of the increasing role of statistics
and mathematical modelling in the pursuit of medicines for
animals and the improved health of species in aquaculture.”
He acknowledged the collaboration, support and work
of Marine Harvest Ltd, Scottish Quality Salmon, the UK
Department for Environment, Food and Rural Affairs and
the Epi-informatics research group at the University of
Strathclyde.
The Dieter Lütticken award, established in 2004, aims to
encourage research into the use of alternative models for
animal testing with significant impact on the development or
production of new animal health products. It is named after
Dr Dieter Lütticken, a committed researcher in microbiology
and virology. He led Intervet’s research and development for
more than 25 years, and retired in 2003 as vice-president and
head of research and development.
The award covers in-vitro models used in research and
development which replace animal testing for licensing
purposes, as well as studies avoiding the use of animals
in efficacy, safety and quality testing in the production of
biologicals and pharmaceuticals for animals.
Intervet said it welcomed submissions from all life-science
research institutions for the next award. This year’s deadline is
November 15. Commercial organisations are excluded.
20 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
Contact Global Communications, Animal Health, Boxmeer,
The Netherlands
Email communications@intervet.com
IRELAND
Improved sea lice strategy
for salmon farms
The Minister for Agriculture, Fisheries and Food, Mary
Coughlan TD, and Minister of State John Browne TD,
recently launched a new strategy to improve pest control on
Irish salmon farms.
The sea louse is the common enemy of both the salmon
farmer and wild fisheries. Monitoring of sea lice on salmon
farms was initiated in 1991 and involves inspecting and
sampling at all fish farms 14 times per year. Treatment is
required where sea lice levels are above target levels.
The strategy intends to build on the existing monitoring
and treatment regime through intensifying and revitalising
the single-bay management approach and making it central
to national policy for sea lice management. The new approach
will incorporate “real time” management of sea lice infestations
on a case-by-case basis.
The strategy also calls for more work to be done to identify
further treatment options, and optimising the use of available
licensed salmon farm sites from a sea lice management
perspective.
A national implementation group comprising representatives
from the Minister’s Department, the industry, BIM and the
Marine Institute, is to be established to review progress made
to reduce sea lice levels and to report within six months with
recommendations on further steps, if any, required to redress
the situation.
CANADA
Public updated on salmon migration
British Columbia’s largest aquaculture company, Marine
Harvest Canada, placed full-page advertisements in the North
Island Gazette and a later edition of the Victoria Times-
Colonist on April 3 to update the public on its success in
minimising the number of sea lice on its farmed fish in
the Broughton Archipelago area during the current out-
migration of wild juvenile salmon.
The advertisement said that that four of its Broughton
farms were now empty of fish. Six other farms showed sea lice
levels of no more than 0.2 lice per farmed fish, well under the
provincial government’s threshold limit of three lice per fish,
at which time fish must be treated to eliminate the lice. The
latest monitoring was completed in early March.
“This data has been available on our website since 2004,” said
the Director of Environmental Compliance and Community
Relations, Clare Backman. “We monitor for sea lice levels diligently
and manage our operations closely to ensure that our fish do not
add to the many other threats that young wild salmon face.”
The ad was the second placed by Marine Harvest Canada to
update the public on its sea lice management plan since the
out-migration began in March. “It is part of our commitment
to being open with those interested in our business about
what we are doing and what results we are attaining. We do
not pretend to have all the answers, but we are committed to
reviewing and supporting science and improving our practices
so that we minimise risk to our salmon and wild stocks.”
Marine Harvest Canada produces 45,000 tonnes of salmon
year-round in farms off Vancouver Island and the central coast.
The company employs 500 people from Duncan to Klemtu.
See www.marineharvestcanada.com/documents/MHC_
sealice_update_April_08. pdf ■
RESEARCH FOCUS

SELECTIVE BREEDING CAN

REDUCE SALMON LICE PROBLEMS
BY BJARNE GJERDE (NOFIMA, NORWAY)
AND SCOTT PEDDIE (AQUACULTURE HEALTH INTERNATIONAL UK)

S
ome salmon of the same species are more susceptible
to lice infestation than others. Utilising this knowledge
in designing selective breeding programmes not only
has the potential to save millions in lost revenue, but also
reduce the infestation pressure of salmon lice among wild
salmonid populations.
These are the findings of a recent Nofima research
project, and follows on from work published by Akvaforsk
researchers in 2005 (Kolstad et al 2005). This initial work
recorded susceptibility to the salmon louse Lepeophtheirus
salmonis in three year-classes of Atlantic salmon with 300
(year-classes 2000 and 2001) and 50 (year-class 2001) full- BJARNE GJERDE
sib families.
The genetic correlations between body weight and lice numbers of
lice were of moderate size, showing that it was possible to improve
both body weight and resistance to L salmonis simultaneously
through selection. Moreover, the genetic correlation between the
numbers of lice recorded in a challenge test and during a natural
infection was very high (See Figure 1).
It was concluded in this initial study that the potential for
improving resistance to sea lice in Atlantic salmon by selective
breeding was good. In addition, as natural infection is highly variable
in time and magnitude, the study recommended that challenge tests
should be used in selective breeding to increase the resistance of
salmon to L salmonis.
Recent Norwegian media coverage has indicated that salmon
lice have become resistant to anti-lice medication. However, one
measure to combat this particular problem is by breeding farmed
SEA LICE
MARINE SALMON FARM
salmon that are more resistant to lice infestation.
“The aquaculture industry and authorities should join forces
to breed a salmon with greater powers of resistance to salmon
lice,” says Nofima (formerly Akvaforsk) senior scientist Bjarne
Gjerde. “Breeding for resistance to salmon lice should be included
as a supplementary measure in the industry’s and authorities’
action plan against lice, in the same way as there is today an
order to delouse as soon as a stipulated number of lice per fish is
exceeded.”
INFESTATION OF WILD SALMON
Although the issue is a complex one, salmon lice from farmed
salmon is one of the threats facing wild salmonids, given that wild
salmon may be infested by lice spread by coastal currents. “The
responsibility to take good care of wild salmonids and the risk that
the lice develop resistance to medication means that it is important
to increase the farmed salmon’s powers of resistance to salmon
lice,” says Gjerde. “Purposeful breeding for a farmed salmon that is
infested by salmon lice to a lesser extent than today will also reduce
the infestation pressure on wild salmonids, as there would be fewer
salmon lice along the coast.”

FIGURE 1: Breeding values for the number of lice per fish of 50 Atlantic
salmon full-sib families under controlled (challenge test with post smolts)
and natural infection (Gjerde et al 2008, data from Kolstad et al 2005)

1.6

1.5
Field test, no. of lice per fish

1.4

1.3
rg = 0.88 ± 0.26
1.2

1.1

1.0

0.9

0.8
45 50 55 60 65 70 75 80 85 90 95
Challenge test, no. of lice per fish

22 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008


FORTHCOMING
FISH AND SHELLFISH EVENTS
THE 1ST INTERNATIONAL CONGRESS ON
HEALTH MANAGEMENT AND DISEASES OF
AQUATIC ANIMALS
Tehran, Iran
January 27-28, 2009
This international meeting will be hosted by the Veterinary Council,
IR Iran, as organiser, with the collaboration of Contemporary
Conference Organisers as the co-organiser. The congress has
the support of the Iranian Veterinary Organisation, Faculty of
Veterinary Medicine of University of Tehran, Iranian Fisheries
Research Organisation, Iranian Fisheries Organisation, Iranian
Veterinary Association, Faculty of Veterinary Medicine of Islamic
Azad University and the Iranian Department of Environment.
The main aim of this congress and exhibition is to create a
dynamic scientific environment for presenting, transferring and
exchanging the latest advanced research findings and scientific
achievements in all health and nutrition management aspects of
aquatic animals.
There will be particular emphasis on diseases, prevention and
treatment, nutrition health management, water quality management
and health management in aquaculture. This will be achieved
through the active participation of Iranian and overseas veterinarians,
researchers, scientists and experts.
See www.ichmda.com
THE 11TH AQUACULTURAL INSURANCE AND
RISK MANAGEMENT CONFERENCE
Grand Villa Argentina,
Frana Supila 14, 20000 Dubrovnik, Croatia
March 26-27, 2009
Aquaculture Underwriting and Risk Management Services Ltd has
▲
CONTINUED FROM PAGE 18
ECONOMICS
Economic losses due to sea lice infestation are substantial in the
salmon industries of the major producing countries. Selective
breeding for increased resistance to salmon lice can potentially
provide significant economic gains in two ways. Firstly, farmed
salmon will require fewer treatments against salmon lice and
secondly, the infestation pressure on wild salmonids will
decrease.
The first is of major economic value to the salmon farming
industry, while the latter is of major value to society. “But it is
important to emphasise that this, as with other traits selected for
in breeding programmes, is a long-term measure,” says Gjerde. “It
will be some years before we reap the big gains, so it is important to
commence this work as quickly as possible.”
ACKNOWLEDGEMENTS
The project is financed by the Research Council of Norway and
Salmo Breed AS. The findings from a fresh project involving
EVENTS
announced the 11th biennial conference. It is targeted at insurance
underwriters, brokers, loss adjusters, surveyors, lawyers, fish farm
risk managers, university experts, investors and all those interested in
insurance, risk management and loss prevention in aquaculture.
Confirmed speakers to date include:
• High risk, low reward: has our industry peaked or simply reached
a plateau?
Speakers: Tom Rutter and Martin Coull
• The use of geographical information systems (GIS) in risk
managing aquaculture.
Speaker: Andreas Siebert
• Federal trout and catfish aquaculture insurance development in
the US.
Speakers: Prof Keith H Coble and Assoc Prof Terry Hanson
• Effects of global warming on low-lying coastal mariculture.
Speaker: Ian Thomas
• A review of the current disease situation in aquaculture.
Speaker: Dr Scott Peddie
See www.conference.aquacultureinsurance.com/index.cfm
2008 FISH HEALTH SECTION – AMERICAN
FISHERIES SOCIETY MEETING
Atlantic Veterinary College, The University of Prince Edward
Island, 550 University Avenue, Charlottetown, Prince Edward
Island, Canada
July 9-12, 2008
The meeting is being hosted by the Atlantic Veterinary College
University of Prince Edward Island, and will mark only the second
time that the Fish Health Section annual meeting has been held in
Canada. We are encouraging our EAFP and other international fish
health colleagues to attend.
Prince Edward Island is easily accessible from Europe, Asia and
South America through Halifax, Nova Scotia and Moncton, New
Brunswick by air on several major airlines.
See http://ocs.vre.upei.ca/index.php/FHS/FHS2008 ■
150 families and financed by the Aquaculture Industry Research
Fund (FHF) and Salmo Breed AS will be available later this
(northern) summer.
Contact Bjarne Gjerde, phone (+47) 93 06 15 41 or email bjarne.
gjerde@nofima.no
REFERENCES
Kolstad K, Heuch PA, Gjerde B, Gjedrem T and Salte R (2005). Genetic
variation in the resistance of Atlantic salmon (Salmo salar) to the
salmon louse Lepeophtheirus salmonis. Aquaculture 247. pp145-151
Gjerde B, Sonesson A, Storset A and Rye M 2008. Selective Breeding
and Genetics – Atlantic salmon. In: Aquaculture – Production of
Aquatic Organisms (2000-2005). Aquaculture Research: From Cage
to Consumption (eds) M Thomassen, R Gudding, B Norberg and
L Jørgensen. The Research Council of Norway, ISBN 978-82-12-
02409-0 (pdf version). ■
JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 23
COMMERCIAL FOCUS
CYTOGENIX ANNOUNCES
AGREEMENT TO DEVELOP
DNA VACCINES
SCOTT PEDDIE, AQUACULTURE HEALTH INTERNATIONAL, UNITED KINGDOM
T
he US-listed company CytoGenix Inc announced on March
5 that it has completed an agreement with Taiwan Cobia
Inc to develop DNA vaccines against fish viruses. Under this
agreement, CytoGenix will develop DNA vaccines against selected
viruses, and TCI will vaccinate the fish and test immunological
responses.
The major diseases and target species of interest include viral
nervous necrosis disease (VNN; Betanodavirus, Nodaviridae) affecting
marine fishes such as parrot fish, groupers, flounders, sea bass,
turbot, and striped jack. DNA vaccines against pathogenic Iridovirus
in grouper (Lymphocystivirus, Megalocytivirus or Ranavirus) are also
being investigated.
“Fish is man’s most important single source of high-quality
protein and provides ~16 percent of the animal protein consumed
by the world’s population (United Nations Food and Agriculture
Organization, 1997),” says the president and chief executive officer
of TCI, Cory Huang.
According to the FAO, aquaculture is one of the fastest growing
food-producing sectors of the world, and it continues to grow more
rapidly than all other animal food-producing sectors. Asia and
the Pacific region contribute over 91.5 percent of the total global
production and 82 percent of the value, Huang said.
“However, aquaculture faces many challenges, especially when it
comes to health management of fish stocks. CytoGenix’ synDNATM
technology has the capability to provide the large quantities of
affordable DNA vaccines necessary to address the need for simpler
and more effective aquaculture vaccination regimens than currently
available.”
CytoGenix’s chief scientific officer, Dr Yin Chen, says DNA
vaccines have already been shown to provide protection against
pathogenic challenges in several animal models, including fish.
“The vaccines carry only a small portion of the infective agent, and
therefore carry almost no risk of inadvertent infection, particularly
when manufactured using CytoGenix’s proprietary cell-free method
(synDNATM),” Dr Chen said.
“Additionally, the prophylactic potential of genetic vaccination
in fish culture is being commercialised, due to the advantages
offered by genetic vaccines over conventional preparations ie, ease
of production, stability, cost and production of vaccines against
organisms which are difficult or dangerous to culture in the
laboratory.”
The president and chief executive officer of Cytogenix, Malcolm
Skolnick, says the synDNATM vaccine platform is highly adaptable.
“These vaccines are based on an expression cassette that may be used
against a large number of pathogens by substituting appropriate
sequences to express antigenic proteins for a given pathogen.
“This agreement represents a beneficial joint effort where
CytoGenix develops a vaccine and our strategic partner tests it and
has the ability to commercialise the results. This agreement with
Taiwan Cobia further exemplifies the versatility of our technology and
provides an avenue for expansion into the worldwide marketplace,”
Skolnick said.
24 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
THE CYTOGENIX TECHNOLOGY
The CytoGenix team has developed a method for manufacturing
large amounts of high quality therapeutic nucleic acid constructs
(synDNATM) which is conducive for the rapid development of
compounds with new sequences of interest. The process offers
significant improvements over traditional bacteria-based
fermentation for plasmid DNA production and polymerase chain
reaction (PCR)-based methods.
It incorporates a cell-free method for producing therapeutic
quality DNA for use as drug substances in both humans and
animals. According to CytoGenix, such an approach offers a number
of advantages, including:
• Minimal hazardous contaminants: Cell-free amplification of
therapeutic nucleic acid has many important benefits, beginning
with a smaller size and less complex version of the plasmid. Under
this system, there is no need for bacterial replication sequences or
selection markers such as antibiotic resistant genes found in the
plasmid DNA backbone. In most cases, this will reduce the size
and weight of the therapeutic product by at least 3000 base pairs.
In addition, the absence of bacteria and growth medium greatly
simplifies the methodology required to purify the manufactured
product. This process employs a simplified chromatography-based
purification procedure developed at CytoGenix. Depending on
the intended application, nucleic acids of various grades of purity
can be produced ranging from 70 percent (ie, no purification)
to about 95 percent pure. This reduces or eliminates the need
for mechanical or chemical purification methods to remove
contaminants such as bacteria cellular debris, endotoxins, RNA or
genomic DNA molecules.
• Robust biological activity: According to CytoGenix, the results
of their experiments have shown that the biological response to
this material (devoid of vector backbone) is similar to traditional
plasmids. Moreover, experiments conducted in several animal
models have shown that linear DNA prepared with CytoGenix
synDNATM process triggers a robust response (immune or
physiological, depending on the application) in treated groups
compared to placebo or plasmid DNA-treated animals.
• Low risk, competitive cost, universal accessibility and fast cycle
time: CytoGenix scientists claim that the entire process is bench-
scale and requires little equipment, space or human intervention
SENIOR SCIENTIST FRÉDÉRIC KENDIRGI, PHD NEXT TO THE HPLC PURIFICATION
STATION FOR SYNDNATM TECHNOLOGY
in comparison to bio-process or bacterial fermentation
manufacturing facilities.
This process easily lends itself to liquid-handling automation,
and a skilled technician can synthesise multi-gram quantities of
this material within a few weeks, while working in a compact,
room-sized facility. Unlike PCR, this process requires only basic
laboratory equipment and is therefore accessible to many facilities
around the world, especially rural settings in the field, where
access to specialised equipment is limited and often prohibitive.
• Improved regulatory profile: A major benefit of using this
cell-free DNA manufacturing technology, again according to
CytoGenix representatives, relates to the regulatory agency review
and compliance perspective. CytoGenix claims that product
cGMP manufacturing procedures detailing methods for cell
collection, processing and cell culture conditions are no longer
necessary, and therefore reduces the level of risk, the amount of
documentation and the amount of required space, as well as QA/
QC and compliance costs.
In terms of production, CytoGenix states that once all the
production parameters are optimised for a given construct,
manufacturing of gram quantities of the desired product can usually
be accomplished within five to 10 working days. Quality control tests
are incorporated for quality assurance and within another week, the
DNA can be released for use. Additional developments may reduce
the lag period between custom optimisation identification and DNA
release.
CytoGenix has entered into an exclusive supply agreement with
GE Healthcare Bio-Sciences Corp to purchase custom kits and
reagents for making large amounts of nucleic acid in accordance
with our proprietary technology. The agreement gives CytoGenix
the exclusive, worldwide right to purchase the necessary reagents
from GE Healthcare for use in producing therapeutic synDNATM.
ABOUT CTYOGENIX
CYTOGENIX MANAGEMENT TEAM
Malcolm Skolnick,
PhD, JD. Chairman/president and chief executive officer
Malcolm Skolnick received his PhD in physics from Cornell
University and his JD from the University of Houston Law Centre.
Prior to joining CytoGenix, Dr Skolnick held academic positions in
the Graduate School of Biomedical Sciences and the School of Public
Health at the University of Texas Health Science Centre at Houston.
He is currently an adjunct professor in the School of Public Health,
where he formerly served as Professor of Technology and Health
Law.
SENIOR LABORATORY DIRECTOR MS HARILYN MCMICKEN AT THE
DENSITOMETRY STATION FOR SYNDNATM
Prior to joining the School of Public Health, Dr Skolnick managed
the Health Science Centre’s Office of Technology Management and
oversaw the university's activities in protecting and licensing its
patent portfolio of technology.
Dr Skolnick held several clinical trials while he was at the Health
Science Centre in Houston in pain management, smoking cessation
and reduction of withdrawal symptoms in drug addiction. He is
a registered patent attorney, a patented inventor and is licensed to
practice law in the state of Texas. Dr Skolnick has been active in
patent prosecution and licensing for selected clients, and has served
as an expert witness in intellectual property, product liability and
accident reconstruction matters.
Greg S Taylor, CPA. Vice-president of finance and administration
and chief financial officer.
Greg Taylor has almost 20 years of investment banking, venture
capital, corporate advisory and entrepreneurial experience and
INFORMATION BOX
Taiwan Cobia Inc, a private company founded in 2004,
was the first company to introduce the concept of organic
aqua-farming in Taiwan. The company is currently adding
production sites in China and South East Asia. TCI also
has collaborative agreements with research institutes and
universities, including Academia Sinica, University of
California at San Diego, National PengHu University, and
National Taiwan University, as well as various government
agencies in Asia and Africa.
CytoGenix Inc is a Houston-based bio-pharmaceutical
company pioneering the new field of gene-based medicine.
It is focused on developing innovative vaccines and
therapeutic products using its three proprietary technology
platforms:
• synDNATM enzymatic, non-bacterial production
methodology
• single stranded DNA (ssDNA) expression, and
• OligogenixTM, an oligonucleotide based anti-bacterial
technology.
CytoGenix currently holds 13 granted patents, two
allowances and 55 additional international or US patent
applications claiming methods and materials in connection
with these platform technologies.
See www.cytogenix.com
JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 25
COMMERCIAL FOCUS
CYTOGENIX ANNOUNCES AGREEMENT TO DEVELOP DNA VACCINES
FREDERIC AND HARILYN NEXT TO TC HOOD (FUNCTIONAL ANALYSIS IN
CULTURED CELLS OF THERAPEUTIC DNA PREPARED WITH SYNDNATM PLATFORM)
has completed over $2BB in transactions. He has also served in
management positions as both a chief executive officer and chief
financial officer.
He was most recently managing director of Monterey Capital
Partners, a private equity investment firm, for over 10 years.
Previously, he served as a vice-president with the investment banking
division of Sumitomo Trust and Banking and with Bunker Hill
Associates, a merchant banking firm. Greg Taylor began his career
with Ernst and Whinney, a big eight accounting firm in Houston,
Texas as a certified public accountant.
He has served on the board of directors of several private
companies and charitable organisations. He has a Bachelor of
Business Administration in accounting and finance from Baylor
University and a Master of Taxation from Baylor University.
Yin Chen, PhD. Chief scientific officer and vice-president of
research and development
Dr Chen earned his PhD in molecular biology and biochemistry
from the University of Maine. Subsequently, he was a post-doctoral
fellow at the Beth Israel Deaconess Medical Centre, a teaching
hospital of the Harvard Medical School. He was briefly employed by
InGene, Inc in Kansas City, MO as a senior research scientist.
Dr Chen joined CytoGenix In 2000 as chief research scientist. He
was promoted to vice-president of research and development in
2001. Dr Chen oversees the research and development team at the
company and is the leading inventor of the company’s proprietary
technologies.
Pam Schertz, CPA, Controller
Pam Schertz has been appointed as interim chief financial officer.
Ms Schertz has served the company as controller since 2003. She
received her BBA in accounting from the University of Houston
and is a licensed CPA in the state of Texas. After establishing her
own practice offering consultant services as a CPA in October 2003,
Cytogenix became her first client.
26 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
THE CYGX SCIENTIFIC TEAM. FROM LEFT: CHIEF SCIENTIFIC OFFICER YIN CHEN,
PHD, SENIOR LABORATORY DIRECTOR MS HARILYN MCMICKEN, BS, MT,
SENIOR SCIENTIST FRÉDÉRIC KENDIRGI, PHD AND SENIOR RESEARCH SCIENTIST
XIN-XING TAN, PHD
Prior to working for Cytogenix, Ms Schertz has had over 15 years
of experience in accounting in various industries, including a casket
manufacturer, an independent power company and an architecture
firm. Management positions have included serving as controller for
a high-end clothier and a civil engineering firm.
Cindee Ewell, PhD, JD.
Vice-president of legal affairs and secretary
Dr Ewell has been with CytoGenix since 2004 and is the company’s
resident corporate and patent counsel. She received her law degree
from the South Texas College of Law, her bachelor’s degree in
microbiology from Cornell University, and her doctorate in
biochemistry and molecular biology from the University of Texas
Health Science Centre at Houston.
She is a registered patent attorney and is licensed to practice law in
the state of Texas. Her experience includes nearly eight years of research
at Baylor College of Medicine, about four years of patent prosecution
and freedom to operate experience in a large law firm environment,
and several years of contract and corporate law experience.
Ms Ewell entered the legal field in 1999 as a biotechnology
technical advisor with a focus on intellectual property. She has since
attained her law degree and has established herself as a patent and
corporate attorney with a specialty in biotechnology.
Key CytoGenix staff
Xin-Xing Tan, PhD. Senior research scientist
Dr Tan earned his PhD in biochemistry from the Chinese Academy
of Science, Shanghai, China. He was an NIH-sponsored post-
doctoral fellow at Rice University, Houston. He has authored or co-
authored 20 scientific published papers. Dr Tan joined CytoGenix in
2002 and is co-inventor of an anti-bacterial technology developed in
the company’s laboratory. He leads the company’s research efforts in
developing novel antimicrobial therapeutics.
Frederic Kendirgi, PhD. Senior scientist
Dr Kendirgi earned his MSc in virology and immunology in 1995 October 2002
from the University of Quebec/INRS-Institut Armand-Frappier, A marvel of biochemical engineering means cells can produce DNA
Canada, and his PhD in biochemistry and molecular biology in 2000 enzyme to attach cancer, New Scientist. January 2001
from the University of Calgary, Canada.
Following four years of postdoctoral studies in cell biology at Conference presentations
Washington University and Vanderbilt University, Dr Kendirgi joined Chen Y, Ji Y and Conrad C. Novel intracellular single-stranded DNA
the CytoGenix research and development team in October 2004. He is a expression system: production of c-raf RNA-cleaving DNA enzyme.
co-inventor of the synDNATM technology and oversees manufacturing Keystone Symposia, Gene Therapy 2001: a gene odyssey. January 6-
operations as well as the synDNATM vaccine platform. 11, 2001, Snowbird, Utah
Chen Y. Novel intracellular single-stranded DNA expression system:
Harilyn W McMicken, BS, MT (ASCP). Senior laboratory director production of c-raf RNA-cleaving DNA enzyme. Tenth International
Harilyn McMicken graduated from Abilene Christian University Conference on Gene Therapy of Cancer. December 13-15, 2001, San
and University of Texas at Galveston. She has done postgraduate Diego, California. (Invited speaker)
work at the University of Texas School for Biomedical Sciences in
Chen Y and McMicken H. Intracellular production of DNA enzyme
Houston, Texas. Prior to her employment, Ms McMicken was senior
by a novel single-stranded DNA expression vector. Fifth annual
research assistant for 23 years in the Department of Paediatrics at
meeting of the American Society of Gene Therapy. June 5-9, 2002,
the Baylor College of Medicine. Her responsibilities and expertise
Boston, Massachusetts
lie in the areas of molecular biology and laboratory and personnel
management. Chen Y. Application of a novel single-stranded DNA expression
vector for gene regulation. Eleventh International Conference
KEY CYTOGENIX SCIENTIFIC PUBLICATIONS on Gene Therapy of Cancer. December 12-14, 2002, San Diego,
Scientific publications California
Kendirgi F, Yun NE, Linde N, Zacks MA, Smith J, Smith J, McMicken Chen Y and McMicken H. Application of novel single-stranded DNA
H, Paessler S and Chen YA. Novel DNA-based vaccine protects expression vector for gene regulation. Keystone Symposia, Drug
mice against lethal infection with H5N1 influenza virus isolate A/ Target Validation Gene Suppression, January 17-22, 2003, Tahoe
Vietnam/1203/04. Human vaccine 2008 (In Press). City, California
Tan X, Knesha R, Margolin W and Chen Y. DNA enzyme generated by Chen Y. Novel Intracellular Single-Stranded DNA Expression System
a novel single-stranded DNA expression vector inhibits expression and its Applications. Third annual conference: RNA in Drug
of the essential bacterial cell division gene ftsZ, Biochemistry 43, Development – RNA as Tool and Target, San Diego, CA. November
2004. pp1111-1117 10-13, 2003. (Invited speaker)
McMicken H, Bates P and Chen Y. * Antiproliferative activity of Chen Y et al. In vivo generation of oligodeoxynucleotides for
G-quartet-containing oligonucleotides generated by a novel single- regulating gene expression. Keystone Symposia, New Advances in
stranded DNA expression system. Cancer Gene Therapy 10 (12)
2003. pp867-869
Chen Y and McMicken H. Intracellular production of DNA enzyme
by a novel single-stranded DNA expression vector. Gene Therapy 10.
pp1776-1780, 2003
Chen Y, Ji Y and Conrad C. Expression of single-stranded DNA in
mammalian cells, Biotechniques 34, 2003. pp167-171
Chen Y. A novel, single-stranded DNA (ssDNA) expression vector
(Review). Expert Opinion on Biological Therapy 2, 2002. pp735-740
Chen Y. Meeting highlights, 10th international conference on gene
therapy of cancer, Expert Opinion on Biological Therapy 2, 2002.
pp443-445
Datta H and Glazer P. Intracelullar generation of single-stranded DNA
for chromosomal triplex formation and induced recombination,
Nucleic Acid Research 29, 2001. pp5140-5147
Chen Y, Ji Y, Roxby R and Conrad C. In vivo expression of single-
stranded DNA in mammalian cells with DNA enzyme sequences
targeted to c-raf. Antisense and Nucleic Acid Drug Development 10,
2000. pp415-422
Other publications
Xin-Xing Tan and Yin Chen. A novel genomic approach identifies
bacterial DNA-dependent RNA polymerase as the target of an
antibacterial oligodeoxynucleotide, RBL1. February 9, 2005
Chen Y and Tan X-X. Oligodeoxynucleotide intervention for the
prevention and treatment of sepsis, April 2004. pp21-23
Chen Y. Novel technologies for target validation. Genetic Engineering
News 23 (11) 2003. pp7-9
21st Century Medicine: Genetic therapy promises dramatic change
from treating symptoms to eliminating the disease. E*Street Journal,
December 2002
Chen Y. Growth of oligo-based drugs, Genomics & Proteomics.

JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 27

LABORATORY FOCUS
THE CENTRAL FISH-HEALTH
LABORATORY IN ISRAEL
BY DR RAMA FALK
KIBBUTZ NIR-DAVID SURROUNDED BY FISH PONDS WITH THE
CENTRAL FISH HEALTH LABORATORY HIGHLIGHTED
T
he Central Fish-Health Laboratory is located in the Jordan
River basin, Beit-Shean Valley, within a rural area where almost
70 percent of Israeli fish farming is centred. CFHL belongs
to the Department of Fisheries and Aquaculture, Ministry of
Agriculture and Rural Development, Israel. It specialises in fish and
shellfish animal health and environmental quality, provides disease
surveillance and diagnostic services, export certifications and offers
advice and guidance on health management and treatment for edible
and ornamental fish growers.
The Central Fish-Health Laboratory addresses the basic
requirements of a functioning fish-diseases laboratory in terms of
fish and water sampling methodology and expertise, and experience
in parasitical, viral and bacterial diseases of aquatic animals, as
well as in histopathology, molecular biology, immunology, vaccine
development and pathogenicity studies.
The CFHL has been the only institute in Israel specialising in
aquatic animal health for many years and is involved in a wide
range of research projects concerning fish health and culture. The
laboratory works with individuals, companies, organisations and
academic institutions to develop and provide health management
services, ensuring high standards of veterinary care and attention.
The CFHL was established in 1941 when aquaculture in Israel
began, and its main objective was research of the toxin-producing
alga Prymnesium parvum. One of its founders, Professor Moshe
Shilo and his team from the Hebrew University of Jerusalem,
then discovered the mode of action of P parvum ichthyotoxin and
developed a bioassay for its quantification and verification.
This research was done in collaboration with the supervisor of
28 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
the CFHL, Shmuel Sarig, who was one of the originators of the
European fish pathology associations (COPRAQ, EAFP).
CFHL’s research priorities are set by the current needs of the fish
industry in Israel, and these needs have changed over time. The
severe shortage of water and land in Israel has lead to intensification
of fish culture from 30kg to 200kg per hectare, and a move to use
relatively brackish water, thus providing suitable conditions for the
occurrence of Prymnesium. These conditions raised the awareness
of the importance of water quality examinations introduced by the
CFHL as a routine test performed within the field as well as in the
lab.
Israel’s location within the main routes of the mass migration
of birds between Eurasia and Africa introduced a vast number of
pathogens into aquaculture. Consequently, the CFHL was involved
in introducing many new treatments and nutrition programmes for
Israeli aquaculture, while remaining cognisant of ecological impact
in accordance with internationally accepted standards.
CFHL’s research focused on parameters of stress, mostly in
tilapia, the main fish raised in Israel. This research, led by the
CFHL’s supervisor for 30 years, Dr Itzhak Bejerano, elucidated the
relationship between farm management, water quality and pathogen
occurrence.
A major research priority of CFHL research was the urgent need to
find a substitute treatment to malachite green, the use of which was
prohibited a number of years ago, for saprolegniasis. Saprolegniasis
infections cause vast economic losses to Israeli aquaculture as a
consequence of the immunological stress experienced by tilapia
during the winter. In the course of this study the whitening agent
blankophor-BA was found to be highly effective in preventing and
treating Saprolegnia infections in both fish and fish eggs.
During recent years, Israel’s ornamentals industry has expanded
significantly, currently constituting eight percent of the exports
of freshwater fish into the European Union, and three percent of
the world ornamentals market. As a consequence, the scope of the
laboratory expanded to encompass export certification and exotic
disease diagnosis as well.
One of the CFHL’s contributions in this field was the first
characterisation of mass mortalities of koi and common carp, and
in the description of the disease, now known as the koi herpes virus.
This research led to the development of a vaccine for the virus (KV3)
and to further research concerning viral diseases.
Research in CFHL is currently focused on developing a vaccine
for the pathogenic bacteria Atypical Aeromonas salmonicida,
finding alternative treatments to the parasites Hexamita sp and
Ichthiophthyrius sp, and further development of a substitute
treatment to saprolegniasis.
When new, infectious micro-organisms are detected, CFHL is
involved in evaluating its significance, isolation and in further studies
concerning the micro-organism. Research collaboration occurs most
often with academic institutes from Israel and abroad, when CFHL
participates in both competitive government and internationally
funded grants or contracts. HISTOLOGY SECTION OF AN EYE OF A RED-DRUM (SCIAENOPS OCCELATUS)
INFECTED WITH NODAVIRUS. NOTE THE EXTENSIVE VACUOLATION IN THE
GRANULAR LAYERS OF THE RETINA. (GIEMSA)
FACILITIES AND STAFF
The laboratory works under QA demands and is preparing for ISO
17025 accreditation. Facilities and hardware include a necropsy and
histology preparation lab, a microbiology lab, bacterial facilities, a
PCR lab and a variety of routine laboratory equipment.
The facilities include a wet laboratory building as well, with
recirculating support systems. All procedures, care and treatment of
fish are done in accordance with the principles of humane treatment
outlined by the Guide for the Care and Use of Laboratory Animals
(Chief Science, the Ministry of Agriculture, Israel) and are approved
by the Committee for Ethical Conduct in the Care and Use of
Laboratory Animals.
The Department of Fisheries and Aquaculture includes two other
facilities:
1 Aquaculture Research Station Dor. Located by the Mediterranean
Sea, the station includes 40 hectares of earth ponds dedicated to
field trials, and a recirculation aquaculture system intended for LIGHT MICROSCOPY OF THE BULBUS ARTERIOSUS OF TILAPIA
intensive culturing research, hatcheries and quarantine, researching INFECTED WITH THE DIGENEAN ASCOCOTYLE SP. SECTION
mainly genetics, nutrition, reproduction, health, environment and SHOWS THE FIBROUS CAPSULE THAT SURROUNDS THE PARASITE
AND DEFENDS IT FROM HOST REACTION (H&E)
broodstock development of fish. It houses 10 staff and students, all
engaged in research.
2 Ginosar Research Station for Intensive Aquaculture. Located
near the Sea of Galilee, the Ginosar station is focused mainly on
nutritional and intensification studies. It has an area of 10 hectares,
including 100 ponds of various sizes used for experiments, with a
staff of four.
Full cooperation between these three facilities enables the
successful resolution of emerging problems concerning fish health.
The research process usually begins in the CFHL, with in-vitro
susceptibility tests and development of fish-models of diseases in
aquariums and small tanks, proceeds to large-scale experiments in
the Ginosar Research Station in larger tanks and small concrete and
earth ponds, and progresses further to controlled field trials within
earth ponds at the Aquaculture Research Station Dor.
Professional staff at the CFHL include:
• Dr Simon Tinman, DVM, a specialist in fish disease
• Margarita Smirnov, an expert in fish pathology and histology
• Nir Froyman, an ornamental fish genetics specialist and laboratory
HISTLOGY SECTION SHOWING EPITHELIOCYSTIS IN PARROT FISH. THE SYMPTOMS
supervisor SHOWN ARE ENLARGED CELLS IN GILL EPITHELIUM FILLED WITH GRANULES. THE
• Esther Parag, a laboratory technician, and HYPERTHROPHIC CELLS OF VARYING SIZES OCCUR ON THE SURFACES OF THE
• Rama Falk, PhD, a microbiologist. GILL LAMELLAE. (TOLUIDINE BLUE)

JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 29

?????????
THE CENTRAL FISH-HEALTH LABORATORY IN ISRAEL
CFHL STAFF FROM RIGHT: NIR FROYMAN, ESTHER PRAG,
RAMA FALK, MARGARITA SMIRNOV AND SIMON TINMAN
SERVICES PROVIDED
CFHL services are essentially dedicated to fish health, and include
diagnosis of the causative agent or etiology of mortality, together
with programmes for preventing or controlling aquatic diseases.
The laboratory promotes the use of high quality diagnostic and
analytical testing, as well as technological innovation to provide
concrete actions and efficient solutions to meet growers’ needs. The
current fee offered by CFHL is subsidised by government contract
and provides all clients with full confidentiality.
DIAGNOSTIC LABORATORY SERVICES
These services encompass a wide range of diagnostic laboratory
testing, including bacteriology, mycology, parasitology, virology,
water quality, limnology, molecular biology (PCR) and descriptive
histopathology. Results and recommendations of parasite infection
studies and water quality analysis are given on the spot, while
bacteriology results are made available within 48 hours and PCR
results within 10 days. Reference samples for both histological
and molecular diagnosis of many diseases of regional significance,
including an archive of bacterial pathogens, are maintained under
appropriate conditions.
DIAGNOSTIC INTERPRETATION AND
CONSULTATION
CFHL provides interpretations of laboratory results and the
significance of the diagnostic findings. In addition, CFHL staff
advise veterinarians and farmers and give recommendations on
management and remediation procedures concerned with water
treatment, sanitation, feeding and pathogen management, all in full
compliance with regulatory authority requirements. These services
include visits of staff members to aquaculture sites (including high-
intensity production facilities) and fisheries.
30 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
CERTIFICATION AND FARM SURVEILLANCE
CFHL collaborates with the Veterinary Services and Animal
Health to examine fish for infectious diseases notifiable to the
Office International Epizooties or OIE. CFHL staff examine lots
of fish intended for export and provide export certification under
authorisation by the Minister of Agriculture. In addition, the staff
conducts regular surveillance for fish producers, interstate zoning
and transfer support and farm management health maintenance
purposes. Export certification is aimed at meeting the demands
of authorities from various countries and is done under strict
regulations.
RESEARCH AND DEVELOPMENT SERVICES
CFHL offers various companies a screening programme to verify
their product’s usefulness for aquaculture. This programme
includes susceptibility tests to various fish pathogens, toxicity tests,
therapeutic-efficacy examinations in fish-models and further field
trials for efficacy determination.
TRAINING AND EDUCATION
CFHL conducts training workshops concerning fish health and
fisheries management and certifies professionals in the fish industry.
Participants include staff of laboratories and fish farms from Israel
and overseas. The research of new, emerging pathogens over the
years was the basis for MSc and PhD projects within the CFHL, in
collaboration with academic institutes. The findings of these projects
have then been applied in the routine work of the laboratory.
CONTACT DETAILS
Central Fish Health Laboratory, Nir-David, Israel. Phone
(972) 4 658 5877, Fax (972) 4 637 4329. Email ramat@moag.gov.il
or see www.vetserv.moag.gov.il/fishery ■
 A healthy underwater world

A clear vision from

Intervet Aquatic
Animal Health

We think globally but have the right products for local use.
Our quality products are led by the Norvax® range.

Our R&D centre in Singapore is dedicated to improving

aquatic animal health in the Asia-Pacific region.

We pledge to work hand-in-hand with you

to help develop and sustain your future.

A world leader in fish vaccines

For information, please contact:

Asia: Intervet Norbio Singapore • Phone: +65 6397 1121 • E-mail: info.aquaINS@intervet.com
Salmonid countries: Intervet Norbio • Phone: +47 5554 3750 • E-mail: info.norbio@intervet.com
Elsewhere: Intervet International • Phone: +31 485 587600 • E-mail: info.aqua@intervet.com • http://www.intervet.com/aah
R126’03b
RESEARCH FOCUS

WHAT’S NEW IN PANCREAS

DISEASES RESEARCH?
BY MARIAN MCLOUGHLIN
AQUATIC VETERINARY SERVICES, BELFAST, UNITED KINGDOM

REPLICATING VIRUS DISINFECTION IS

EFFECTIVE AGAINST SAV

PD VIRUS
P
ancreas disease (PD) remains one of the most serious
and economically important infectious diseases of farmed PD virus is classified as a salmonid alphavirus (SAV), and
Atlantic salmon (Salmo salar L) in Europe. Over the last molecular analysis now indicates that there are at least six
few years our knowledge and understanding of the PD virus and known sub-types, with very interesting geographical and species
the disease has been expanded considerably, due to the efforts and distribution. See Table 1.
collaboration of excellent research scientists, veterinarians and SAV 3 is the only sub-type found to date in Norway and can
the salmon industry. This effort has been coordinated and greatly cause disease in both Atlantic salmon and sea-reared rainbow
assisted by the formation of the Tri Nation PD Research Group. trout (Oncorhynchus mykiss [Walbaum]). SAV 2 has primarily
The aim of this article is to summarise the new and important been associated with disease in freshwater rainbow trout and
information in pancreas disease research. For a recent review article was only recently introduced to the United Kingdom K. SAV 2,
see Alphavirus infections in salmonids – a review. MF McLoughlin as distinct from freshwater strains, has recently been recognised
and DA Graham, Journal of Fish Diseases, September 2007, Volume in marine Atlantic salmon in Scotland. SAV 1, 4 and 5 are present
30 Issue 9, Special Alphavirus issue. pp509-572 in Ireland and Scotland.
Further work to compare the pathogenicity of these sub-types is
TABLE 1: Geographical and Species Distribution of SAV Isolated in Europe planned in the near future. Interestingly, there is regional separation
of sub-types in Ireland with SAV 1 in Connemara (west) and SAV
SAV SUBTYPE SPECIES COUNTRY
4 in Donegal (north). Given that smolts from one region have
SAV 1 Atlantic salmon Ireland (Mainly Connemara) regularly been transferred to the other, it suggests that vertical
Scotland
transmission has not been a major source of infection between
SAV 2 Rainbow trout France
England & Scotland
these regions (Fringuelli et al 2008 submitted).
Italy
Spain
Germany* presumed SAV2
SAV SURVIVAL
Atlantic salmon Scotland A major gap in the knowledge of SAV was its survival outside
the fish and response to temperature, pH and disinfection,
SAV 3 Atlantic salmon Norway Marine only
Rainbow trout Norway which are very important in the overall control of the disease
SAV 4 Atlantic salmon Scotland
spread. SAV was found to be rapidly inactivated in the presence
Ireland of high levels of organic matter at 60˚C at pH7.2. It is also
SAV 5 Atlantic salmon Scotland
inactivated at pH4 (acid) and pH12 (alkali) at 4˚C, suggesting
that composting, ensiling and alkaline hydrolysis would all be
SAV 6 Atlantic salmon Ireland
effective at inactivating the virus in fish mortalities and waste.

32 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008

 TABLE 2: Diagnostic Panel for Confirmation of PD
A number of common TEST PER ACUTE ACUTE SUB ACUTE CHRONIC CARRIER
disinfectants are effective against DPI 0–7 0 + 14 14 > 21 21 > 42 42 > ?

SAV if used at the recommended Signs $ Appetite Casts Morts Runts

Virus Serum, Serum, Serum,

concentration Heart Heart Heart
– –

Histology N/A + Pancreas + Pancreas + Heart ?

+ Heart + Heart + Muscle
Virus survival was shown to be inversely related to temperature,
IHC N/A + Pan only – – –
ie, the higher the temperature the lower the survival rate, and to
be reduced by the presence of organic matter. Conversely, at RTPCR + + + + +/–
low temperatures the virus survived a relatively long period of Serology – – – + +
up to 61 days in sterile sea water, suggesting that SAV could be
transmitted between farms by currents and tidal flows at low remove inhibiting organic matter. (Graham et al 2007, Journal
water temperatures. of Fish Diseases 30. pp269-277)
Pump-ashore systems would also be at risk unless effective
disinfection procedures are in place. Survival in fresh water is markedly PD DIAGNOSIS
less than in sea water. SAV2 is reported to have lower pathogenicity at The three pillars of disease diagnosis are
10˚C compared with 14˚C, which may explain why SAV infections at • history
low temperature may be predominantly sub-clinical. (Graham et al • clinical signs and gross pathology, and
2007, Journal of Fish Diseases 30. pp533-543) • confirmatory tests
– histopathology
DISINFECTION – serology
A range of commercially available disinfectants, including Virkon – virus isolation/detection (PCR)
S, Virex, Halamid, Buffodine and FAM 30 have been tested using
a protocol based on the BS EN 14675 standard for efficacy EPIDEMIOLOGY AND HISTORY
against SAV. The peroxygen disinfectants Virex and Virkon S Increasing factual information is being gathered from field
were both effective under a range of test conditions with efficacy surveillance and outbreaks in Norway, Scotland and Ireland.
maintained even at 4˚C, and when contact times were reduced to Taksdal et al 2007 have written a comprehensive description of
five minutes, even in the presence of organic matter. PD in Atlantic salmon and rainbow trout in Norway, where the
Halamid and FAM 30 were also effective, but were adversely incidence and geographical spread of PD is increasing each year
affected by the presence of organic matter. Buffodene, an iodophor (Taksdal et al 2007 30. pp545-558. [See also maps 1 and 2]).
disinfectant widely used for egg disinfection, was adversely affected Individual salmon farming companies have interrogated their
in this study by the presence of organic matter, as has previously historic and current data for trends and clues as to the source of
been reported for other fish viruses. Both Virex and FAM 30 were infection and factors that may be contributing to the prevalence
shown to be effective if used in sea water, while Virkon S and and severity of PD on their sites.
Halamid produced precipitates when mixed with sea water, and as In Scotland, a significant sub-clinical pattern of PD disease
a result their efficacy against SAV was not tested in seawater. has been revealed, indicating that infection is much more
In conclusion, a number of common disinfectants are effective widespread than observed disease, and that poor performance in
against SAV if used at the recommended concentration, for an
adequate time and after thorough cleaning of all surfaces to CONTINUED ON PAGE 35

FIGURE 1. THE SPREAD OF PD FROM 2004 TO 2007 IN WESTERN NORWAY

MAP 1:
DISTRIBUTION OF MAP 2: DISTRIBUTION OF
CONFIRMED PD CONFIRMED PD OUTBREAKS
OUTBREAKS IN 2004 UP TO 01/12/2007. SOURCE
MATTILSYNET

2004

JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 33

BOOK REVIEW
AQUACULTURE BIOSECURITY:
PREVENTION, CONTROL AND
ERADICATION OF AQUATIC
ANIMAL DISEASE
EDITED BY A DAVID SCARFE, CHENG-SHENG LEE AND PATRICIA J O’BRYEN. BLACKWELL PUBLISHING, 2006. PP182. ISBN: 978-0-
8138-0539-9
D
isease in aquaculture is responsible
for widespread and often
economically debilitating losses
and has implications for stock welfare,
international trade and consumer
perception and purchasing patterns. As a
consequence, aquaculturists, veterinarians,
policy-makers, scientists and a range of
other stakeholders have become increasingly
aware of the need to develop a coherent
and integrated approach to aquatic animal
disease prevention and management. It is
against such a backdrop that the theory
and practice of biosecurity has gained
increasing prominence in recent years.
During the World Aquaculture Society’s
annual conference in Hawaii in March 2004,
the Oceanic Institute and the American
Veterinary Medical Association brought
together a diverse group of stakeholders
for a three-day session entitled Aquaculture
Biosecurity 2004. The aim of this session
was to present and assess the range of
approaches being taken globally, and
then to identify the key steps necessary to effectively implement
biosecurity programmes in the production environment. Aquaculture
Biosecurity is a compilation of twelve of the 30 presentations given
at the stakeholder session.
Chapters one and two provide an overview of the biosecurity
approaches undertaken at the international level by focussing
on the World Organisation for Animal Health, the World Trade
Organisation and the Codex Alimentarius Commission and their
respective standards aimed at eliminating trans-boundary disease
transmission.
Chapter three presents an interesting and informative example
of a regional approach to aquatic animal health management. In
this chapter the scope and utility of
programmes designed by the Network
of Aquaculture Centres in Asia-
Pacific is presented as an example of
effective regional co-operation. This is
followed by a comprehensive and well-
written account of the development
and functioning of Canada’s national
Aquatic Animal Health Programme,
using relevant case studies to highlight
the need for an overarching and all-
encompassing approach to biosecurity.
The national dimension is continued
A. DAVID SCARFE
in chapter five, where the United States
34 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
Fish and Wildlife Service’s aquatic animal
health policy is discussed, with particular
emphasis on the innovative approaches
to disease management contained therein.
A detailed account is presented of the
plan’s risk assessment procedures, which
were developed to both standardise and
optimise the decision-making process
pertaining to aquatic animal relocation.
The situation in the USA makes for an
interesting discussion point and is explored
in detail in this book. That many states
have no regulations, while others address
only salmonids, provides an entry point to
showcase Wisconsin’s veterinary approach
to fish health, the topic of chapter six.
Wisconsin’s approach is an interesting one,
given that it includes non-salmonids and
deals with fish farm registration, certification
of veterinarians, health standards for fish
introduced into public waters and the
issuing of fish import permits. Moreover,
the programme evolved as a result of an
effective working relationship between the
state government, aquaculturists and veterinarians, and is therefore
an important example of multi-stakeholder co-operation.
Chapter seven makes a coherent and succinctly presented case
for a harmonised system of accreditation to ensure diagnostic data
and test result validity in aquatic animal health diagnostic systems.
This chapter forcefully promotes that valid data and test results
are a critical pre-requisite for effective biosecurity planning and
implementation.
Chapter eight provides an excellent and in-depth review of
the types of disinfectants applicable to aquaculture, as well as
stressing the need for disinfection as an integral component of
effective biosecurity at all levels. Information on the efficacy and
characteristic of commonly used disinfectants, fish pathogen
classification and transmission/susceptibility characteristics of
important fish pathogens is presented clearly and helpfully in
tabular format and makes an excellent point of reference for the
interested reader. Moreover, the “example disinfection protocols”
are another informative reference point and a welcome addition to
a well-rounded chapter.
The ninth chapter highlights the importance of aquatic animal
health surveillance in effective policy making, not only for disease
control but also for quarantine and health certification.
Chapter 10 deals with the key issue of biosecurity at farm level
and how to create the optimum environment and “state of mind”
among farm workers and managers to significantly minimise risk
in aquaculture facilities. This chapter succeeds in emphasising that
effective farm-level procedures are critical in the process. However,
it would have been helpful to see some of the ideas presented in this
chapter developed further.
For example, how can the latest advances in business communication
theory be utilised and developed to inculcate a sense of biosecurity
awareness? What are the motivational models that can best be
applied to the aquaculture environment to enable staff at all levels in
the enterprise to promote and foster “good practice?” It seems to me
that a more detailed and nuanced approach can be formulated, using
human resource management practices, to address this issue and to
develop guidelines for farm managers.
The book closes with two case studies of diseases affecting the
salmon industry: infectious haematopoietic necrosis and infectious
salmon anaemia. Such case studies are certainly helpful in comparing
▲
CONTINUED FROM PAGE 33
the absence of mortality may be related to chronic sub-clinical
infection (Graham et al 2006, Diseases of Aquatic Organisms 72.
pp193-196).
Recent epidemiological results from Ireland revealed that
mean PD-related mortality was 18.8 percent in 2003 and 14.8
percent in 2004, with an estimated loss of growth of 11.8 percent
over the two years. The risk factors associated with PD outbreaks
included positive farms in the area, a history of PD on the site,
more than 250,000 fish on the site and high lice burdens (Rodger
and Mitchell 2007 30. pp157-167).
PD LABORATORY TESTS
Various laboratory tests have been developed and tested in the
past few years and will aid the confirmation of a PD outbreak.
Serology and RT-PCR have also been successfully used as
surveillance tools enabling the identification of acute and
chronic infections. Some companies now carry out routine
serology testing in advance of the risk period(s) for PD as part
of their overall health plan, and have found the early warning
system where viraemia is detected in pre-clinical cases to be very
useful in managing PD outbreaks.
Antibodies develop two to three weeks post infection at 12-
14˚C and have been found to persist up to harvest and are a
good retrospective test for PD infection. Real-time PCR has
also proven to be a very sensitive tool for detecting viral nucleic
acid, but this signal does not persist as long as the presence
of antibody (Graham et al 2005, Journal of Fish Diseases 28.
pp373-379). Recent work indicates that pooled sera can be used
effectively for detecting circulating SAV activity as individual
samples, thus making population screening more cost-effective.
The work on PD survival has also indicated that transporting
sera for virus isolation should be at 4˚C to prevent false negative
results.
Table 3 summarises the test and samples required to confirm a
diagnosis of SAV infection and to differentiate them from similar
pathologies.
REFERENCES
Alphavirus infections in salmonids – a review MF McLoughlin
and DA Graham. Journal of Fish Diseases, September 2007,
Volume 30 Issue 9, Special Alphavirus issue. pp509-531
Fringuelli E, Rowley HM, Wilson JC, Hunter H and Graham
DA 2008. Phylogenetic analyses of partial E2 and ns P3 gene
nucleotide sequences and molecular epidemiology of European
salmonid alphaviruses (SAV). Submitted
Graham DA, Jewhurst VA, Rowley HM, McLoughlin MF, Rodger
and contrasting approaches and make the case effectively for
pathogen-specific strategies where necessary.
Overall, Aquaculture Biosecurity: prevention, control and
eradication of aquatic animal disease, is an indispensable source
of detailed information for biologists, veterinarians, policy makers
and aquaculturists. The editors are to be congratulated for bringing
together so effectively and succinctly a diverse range of topics
presented by such a distinguished list of contributors. The result
is a well-balanced mix of legislative, scientific and management
information that both complements and augments the existing
literature on this most important of topics.
Dr Scott Peddie
Aquaculture Health International ■
TABLE 3:
SUGGESTED SAMPLES FOR DIFFERENTIAL DIAGNOSIS OF PD, HSMI & CMS
TEST SERUM HEART KIDNEY GILL PANCREAS MUSCLE
Virus R R R R
SAV RTPCR R R
SAV
Antibody R
Histology R R R R R
Serology and RT-PCR have
also been successfully used
as surveillance tools enabling
the identification of acute and
chronic infections.
H and Todd D 2005. Longitudinal serological surveys of Atlantic
salmon Salmo salar L using a rapid immunoperoxidase-based
neutralisation assay for salmonid alphavirus. Journal of Fish
Diseases 28. pp373-379
Graham DA, Jewhurst H, McLoughlin MF, Sourd P, Rowley HM,
Taylor C and Todd D 2006a. Sub-clinical infection of farmed
Atlantic salmon Salmo salar L with salmonid alphavirus – a
prospective longitudinal study. Diseases of Aquatic Organisms
72. pp193-199
Graham DA, Cherry K, Wilson CJ and Rowley HM 2007.
Susceptibility of salmonid alphaviruses to a range of chemical
disinfectants. Journal of Fish Diseases 30. pp269-277
Graham DA, Staples C, Wilson CJ, Jewhurst H, Cherry K, Gordon
A and Rowley HM 2007. Biophysical properties of salmonid
alphaviruses; influence of temperature and pH on virus survival.
Journal of Fish Diseases 30. pp533-277
Rodger H and Mitchell S 2007. Epidemiological observations
of pancreas disease of farmed Atlantic salmon Salmo salar L in
Ireland. Journal of Fish Diseases 30. pp157-167
Taksdal T, Olsen AB, Bjerkås I, Hjortaas MJ, Dannevig BH,
Graham DA and McLoughlin MF 2007. Pathology of pancreas
disease (PD) in farmed Atlantic salmon Salmo salar L and
rainbow trout Oncorhynchus mykiss W in Norway. Journal of
Fish Diseases 30. pp545-558
Special note: Regular updates on pancreas disease will be posted on
a new, dedicated PD section at www.aqua.intervet.com ■
JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 35
ORNAMENTALS FOCUS
A BASIC OVERVIEW OF
MULTICELLULAR PARASITIC
INFECTIONS
BY GERALD BASSLEER, BASSLEER BIOFISH, BELGIUM
This article is based on material contained in The New illustrated Guide to Fish Diseases by Gerald Bassleer
ISBN 90-807831-2-9. See www.bassleer.com
CARASSIUS AURATUS SARASSA. PARASITIC AND CARASSIUS RED ORANDA. PARASITIC INFECTION. RED
BACTERIAL INFECTION. SARASSA WITH NUMEROUS SKIN ORANDA WITH SMALL, WHITE SPOTS ON THE HEAD CAUSED
FLUKES AND BACTERIAL INFECTION ON SKIN AND TAIL BY SKIN FLUKES (GYRODACTYLUS)
T
he most common multicellular parasitic infections in ornamental,
tropical and pond fish are skin and gill flukes. External infections
are caused by skin and gill flukes (Monogenea or monogenean
trematodes) Gyrodactylus species and Dactylogyrus species are
especially common in aquarium fish. However, various other species,
similar to these two, also occur.
GYRODACTYLUS (SKIN FLUKE)
This parasitic fluke quickly reproduces itself on the skin and because
it is live-bearing, the young flukes (up to three at a time) are released
from the adults on the skin. It may also be found in the gills and
can readily be identified in a skin scraping under the microscope.
The lower the temperature the slower skin flukes will develop and
reproduce. Small numbers usually do not cause any harm, except in
small or juvenile fish.
In terms of clinical presentation, there is usually excess
slime formation on the skin, the fish scrape over substrate
or against objects, red patches or lesions can be observed,
with secondary bacterial infection after damage by the skin
flukes. If the parasites have also infected the gills, the opercula
will be flared open and the fish will be breathing rapidly. As
skin flukes are usually accompanied by bleeding patches on
the skin caused by physical damage or bacterial infection, the
parasitic infection is often “masked”. Moreover, morbid fish may
be affected by other parasitic infections, such as Ichthyobodo or
Spironucleus.
Treatment with Fenbendazol (200mg/100 l) is often effective, as
is Trichlorfon, Flubendazol, Formalin, Mebendazol, Praziquantel or
salt. In the case of severe infections, an anti-bacterial (and perhaps
another anti-parasitic) medicine will have to be used to combat
secondary infections. A second treatment will also be necessary
within seven days of administration of the first treatment. Note,
36 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
CARASSIUS RED ORANDA. PARASITIC INFECTION.
RED ORANDA WITH A CRATER IN THE HEAD
CAUSED BY SKIN FLUKE (GYRODACTYLUS)
DAMAGE
however, that there are strains of skin flukes that have already
become resistant to certain medications.
DACTYLOGYRUS (GILL FLUKE)
This parasite mainly occurs on the gills, but it can also occur on the
head and skin. Reproduction occurs by means of eggs that develop
in the substrate (or in the filter), so that the entire holding pond (and
its filter) will have to be treated. If the water temperature is low, the
larvae do not emerge from the eggs until several weeks or months
have passed. This should be taken into account in the spring when
the water warms up and the gill flukes may appear. These parasites
(usually 1mm) can be easily identified by examining a skin or gill
scraping under the microscope.
In terms of clinical presentation, the fish scrape over the substrate
or against objects, they are emaciated, breathe rapidly and have flared
open opercula. A small number of parasites may be present on the
fish without causing any damage; relatively large numbers of them
are required to cause a clinically significant infection. Infection can
cause physical damage, which in turn leads to secondary bacterial
infections that can kill affected fish.
Praziquantel (250mg/100 l for three days) is one of the most
effective medicines currently available. It is recommended that
treatment be repeated for three weeks to ensure that all larvae
have been killed; the filter must also be treated as it may harbour
larvae. Moreover, all of the sick fish can be removed from the
aquarium or pond and treated in a separate tank. Over this
period the flukes in the aquarium/pond will die in the absence
of a host. However, at low temperatures the larvae take a very
long time to hatch (at <5˚C this may even take months), or
they emerge from the eggs as soon as the temperature starts to
rise. This should be taken into account when treating ponds,
especially in the spring. ■
CARASSIUS AURATUS GOLDFISH. PARASITIC AND BACTERIAL
CARASSIUS AURATUS GOLDFISH. PARASITIC INFECTION. EMACIATED
INFECTION. GOLDFISH WITH ULCER CAUSED BY SKIN FLUKE
GOLDFISH WITH GILL FLUKE (DACTYLOGYRUS) INFECTION
(GYRODACTYLUS) AND BACTERIAL INFECTION

CARASSIUS REDCAP ORANDA.

PARASITIC AND BACTERIAL INFECTION. CARASSIUS BUBBLE ORANDA EYE.
EMACIATED REDCAP WITH GILL FLUKES AND PARASITIC INFECTION. IRREGULAR SWIMMING BUBBLE EYE WITH RESPIRATORY
SMALL RED PATCHES (BEGINNING OF A BACTERIAL INFECTION) PROBLEMS AND EXTENDED OPERCULA CAUSED BY GILL FLUKE INFECTION

CYPRINUS CARPIO KOI. PARASITIC AND BACTERIAL INFECTION. KOI WITH

SEVERE GILL FLUKE INFECTION ALSO AFFECTING THE HEAD WITH SECONDARY
BACTERIAL INFECTION

CARASSIUS ORANDA LIONHEAD. PARASITIC AND BACTERIAL

INFECTION. LIONHEAD WITH GILL FLUKE INFECTION AND BLEEDING
UNDER SKIN CAUSED BY BACTERIAL INFECTION

CYPRINUS CARPIO KOI. KOI WITH SUNKEN EYES, EXTRA MUCOUS

ON HEAD AND A DAMAGED GILL COVER. GILL FLUKE INFECTION

CYPRINUS CARPIO KOI. KOI WITH

EXTENDED GILL COVER CAUSED
BY INFECTION WITH GILL FLUKES
(DACTYLOGYRUS)
VA C C I N E S T A T I S T I C S
This column, focusing on monthly Norwegian commercial fish vaccine statistics, is
provided by PHARMAQ. See www.pharmaq.no for monthly updates.

PHARMAQstatistics
Fish vaccines in Norway - March 2008
Oil based vaccines for salmon and trout Record breaking vaccination of salmon in March
March 2008 March 2007 Last 12 months
Doses Share Doses Share Doses Share In March 12.1 million salmon were vaccinated. In comparison
2 diseases 0 0,0 % 110 000 1,6 % 3 590 000 1,5 % to March 2007 the similar number of vaccinated salmon was
3 diseases 1 000 000 7,9 % 380 000 5,5 % 3 585 000 1,5 % 6.5 million. Over the first quarter 2008 as much as 52.5 million
4 diseases 1 040 000 8,3 % 380 000 5,5 % 18 810 000 7,9 % salmon were vaccinated. This constitutes an increase of 24 %
5 diseases 10 545 000 83,8 % 5 980 000 87,3 % 213 295 000 89,1 %
compared to the same period last year. The rolling long-term
Sum 12 585 000 6 850 000 239 280 000
Salmon 12 085 000 96,0 % 6 450 000 94,2 % 223 775 000 93,5 %
trend over the last 12 month period now shows us that nearly
Trout 500 000 4,0 % 400 000 5,8 % 15 505 000 6,5 % 240 million salmon and trout are being vaccinated.
PD vaccines 730 000 420 000 23 165 000
In March 2.4 million vaccine doses were used for farmed cod.
This is at the same level as in March 2007. On the other hand,
Oil based vaccines for salmon and trout this month more that 2 million cod were vaccinated with an oil-
100,0 %
based injection vaccine. Since August 2007 there has been a
90,0 % nice development in the long-term trend of cod vaccination.

80,0 %
According to our information, approx. 700.000 doses of a
70,0 %
single PD vaccine (against pancreas disease), were
60,0 % administered to salmon in March. Re-vaccination with another
vaccine is needed to add protection against other pathogens.
50,0 %

40,0 %

30,0 %

20,0 %

10,0 %

0,0 %
March 2008 March 2007 Last 12 months

2 diseases 3 diseases 4 diseases 5 diseases

Doses of oil based vaccines for salmon and trout Doses of vaccines for
against different diseases - 12 months rolling salmon and trout - 12 months rolling

250 250
Millions
Millions

200 200

150 150
Doses

Doses

100 100

50 50

0 0

2 diseases 3 diseases 4 diseases 5 diseases Salmon Trout - oil based Trout - water based

Oil and water based vaccines for cod

March 2008 March 2007 Last 12 months
Doses Doses Doses
Sum 2 425 000 2 340 000 21 335 000 Definitions

Doses* of oil and water based vaccined for cod 2 diseases = furuculosis and vibriosis
12 months rolling 3 diseases = 2 diseases + coldwater vibriosis
4 diseases = 3 diseases + winter sore
5 diseases = 4 diseases + IPN
24
Millions

PD vaccines = vaccines against pancreas disease. Smolt needs re-

22 vaccination with another vaccine to add protection against other pathogens

20
Last 12 months value is the sum of doses used the previous 12 months.

18 The 12 months rolling graphs shows the trend by using the 12 months values
Doses

over a period - here we follow the trend for the last two years
16
* One dose administered to cod is defined as 0.1 ml of vaccine.
It is important to be aware that the calculated number of vaccinated fish
14
shows an over estimate because immersion vaccines are included in the
calculation.
12

10

38 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008

PHARMAQstatistics
Fish vaccines in Norway - April 2008
Oil based vaccines for salmon and trout The vaccination of salmon generation S1 2008
April 2008 April 2007 Last 12 months has passed total S1’s put to sea in 2007
Doses Share Doses Share Doses Share In April 5.2 million salmon were vaccinated against 3.4 million
2 diseases 30 000 0,6 % 0 0,0 % 3 620 000 1,5 % in April 2007. If we consider vaccination of S1’s 2008 from
3 diseases 300 000 5,7 % 75 000 2,2 % 3 810 000 1,6 % October 2007, the number vaccinated by end of April 2008 is
4 diseases 670 000 12,8 % 90 000 2,6 % 19 390 000 8,0 % 119 million salmon. The total S1 generation 2007 was
5 diseases 4 245 000 80,9 % 3 235 000 95,1 % 214 305 000 88,9 %
estimated to 115 million (ref Kontali Analysis). If part of the
Sum 5 245 000 3 400 000 241 125 000
Salmon 5 215 000 99,4 % 3 000 000 88,2 % 227 300 000 94,3 %
vaccination in September 2007 is included into the S1 2008
Trout 30 000 0,6 % 400 000 11,8 % 13 825 000 5,7 % generation we assume that a total of 125-130 million will be
PD vaccines 715 000 0 23 880 000 put to sea as one-year smolt in 2008.

It was a clear reduction in the vaccination of cod, falling from

Oil based vaccines for salmon and trout 2.6 million doses in April 2007 to 990.000 doses this month.
100,0 %
This variation has been seen earlier, and it is too early to
90,0 % predict any further development for 2008.

80,0 % According to our information, approx. 700.000 doses of a

single PD vaccine (against pancreas disease), were
70,0 %
administered to salmon in April. Re-vaccination with another
60,0 % vaccine is needed to add protection against other pathogens.
50,0 %

40,0 %

30,0 %

20,0 %

10,0 %

0,0 %
April 2008 April 2007 Last 12 months

2 diseases 3 diseases 4 diseases 5 diseases

Doses of oil based vaccines for salmon and trout Doses of vaccines for
against different diseases - 12 months rolling salmon and trout - 12 months rolling

250 250
Millions
Millions

200 200

150 150
Doses

Doses

100 100

50 50

0 0

2 diseases 3 diseases 4 diseases 5 diseases Salmon Trout - oil based Trout - water based

Oil and water based vaccines for cod

April 2008 April 2007 Last 12 months
Doses Doses Doses
Sum 990 000 2 605 000 19 720 000 Definitions

Doses* of oil and water based vaccined for cod 2 diseases = furuculosis and vibriosis
12 months rolling 3 diseases = 2 diseases + coldwater vibriosis
4 diseases = 3 diseases + winter sore
5 diseases = 4 diseases + IPN
24
Millions

PD vaccines = vaccines against pancreas disease. Smolt needs re-

22 vaccination with another vaccine to add protection against other pathogens

20
Last 12 months value is the sum of doses used the previous 12 months.

18 The 12 months rolling graphs shows the trend by using the 12 months values
Doses

over a period - here we follow the trend for the last two years
16
* One dose administered to cod is defined as 0.1 ml of vaccine.
It is important to be aware that the calculated number of vaccinated fish
14
shows an over estimate because immersion vaccines are included in the
calculation.
12

10

© PHARMAQ AS 2008
RESEARCH FOCUS

SPREAD AND COLONISATION

OF ALIEN FISH SPECIES IN OPEN
WATERS: A RELIABLE INDICATOR
OF AQUATIC HEALTH
BY AK SINGH AND WS LAKRA, NATIONAL BUREAU OF FISH GENETIC RESOURCES, LUCKNOW, INDIA, AND
ARVIND MISHRA, UP STATE FISHERIES DEPARTMENT, VARANASI, INDIA

N
ew strains and varieties of alien fish species have been
introduced in several countries, including India (DAIS 2004,
Sena De Silva et al 2006, Singh and Lakra 2006), to compete
on the world market. However, environmental, socio-economic and
biodiversity issues are important considerations for regulating the
importation of alien fishes in India or in other countries.
The introduction of alien fish raises questions about how best to
meet food demands and preserve environmental quality. Four broad
categories exist for ecological impacts of alien fish species. They are:
• basic species interactions such as predation and competition
(DIAS 2004, Singh and Lakra 2006)
• genetic impacts (Senanan et al 2004, Mabuchi et al 2008)
• disease impacts (Arthur 2005) and
• habitat alteration (IUCN/SSC 2000).
Alien species are fundamentally indicators of biological integrity
in two aspects. Firstly, the spread of alien fish species in places
other than where it was originally introduced (ie, post-introduction
expansion). Secondly, alien fish species have been associated with the
decline in or extirpation of native fish in a range of systems because
of predation, competition and/or transmission of disease.
In recent years, many alien fish species have been introduced
into India illegally or otherwise (Table 1). These alien species have
escaped into streams and rivers by human activity and aquaculture
diversification efforts and are likely to be susceptible to invade
new environments and ecosystems. Naturally populating alien fish
species have therefore been considered as an indicator of degraded
stream conditions or the poor health of the aquatic ecosystem.
Extensive studies have demonstrated that five to 45 percent exotic
fish were present in river stretches and streams of Ganga and Yamuna
in Uttar Pradesh, India. The maximum contribution of commercial
catch was constituted by alien tilapia (Oreochromis niloticus) and
common carp (Cyprinus carpio), while there was stray occurrence of
other alien fish species like grass carp, silver carp and bighead carp
or even African catfish Clarias gariepinus.
Tilapia (Oreochromis niloticus) and common carp (Cyprinus
carpio) have formed breeding populations and contributed a large
percentage of the exploited stock in the river. These alien fish species
have started competing for food and space in riverine stretches,
showing diet overlap with native and endemic fish species and thus
environmental changes have been perceived (Table 2).
Thus it may be expected that riverine ecosystems consisting of
various natural foods in different niches of the river is important in
diversified zones that could be homogenised by a feral population
of alien fish species. Unethical over-fishing, the use of chemicals,
poisons and dynamite, a wide array of prohibited fishing methods
and habitat destruction of natural spawning and breeding grounds

TABLE1: Alien food fishes of common occurrence in Indian freshwater aquaculture

COMMON NAME SPECIES YEAR SOURCE OF REASON OF SPREAD INTO NATURAL WATER BODIES
INTRODUCTION INTRODUCTION
Mozambique Orechromis 1952 Indonesia Aquaculture Reservoirs of southern part, lakes, wetlands of West
tilapia mossambicus 1962 Bangkok Bengal and Assam, back waters of Kerala, river
1985 Sri Lanka stretches of northern plain.
Bangladesh
Nepal
Nile tilapia Oreochromis niloticus 1987 Thailand Aquaculture and sewage River stretches of Ganga and Yamuna in Uttar Pradesh
Israel fed fisheries
Common carp Cyprinus carpio 1939 Sri Lanka Aquaculture (Composite Most of the river stretches, reservoirs and lakes.
(Scale carp) communis 1957 Bangkok Fish Culture) Recently formed commercial catch in Yamuna and
Ganga in UP
Mirror carp Cyprinus carpio 1939 Sri Lanka, Aquaculture in high Yamuna, Jhelum, Beas, Satluj
specularis 1957 Bangkok altitudes
Grass carp Ctenopha- 1959 Japan, Aquaculture Reservoirs and few river stretches
ryngodon Hongkong (Composite Fish Culture)
idella
Silver carp Hypophthalmichthys 1959 Japan Aquaculture (Composite Reservoirs, lakes and few river stretches
molitrix Hongkong Fish Culture)
Bighead Aristichthys nobils 1987 Possibly Aquaculture Reservoirs, lakes and few river stretches
Bangladesh &
Nepal
African catfish Clarias gariepinus Not known Neighbouring Aquaculture Few reservoirs, Yamuna and occasionally in
countries Godawari river
Sutch catfish Pangasius sutchi Not known Neighbouring Aquaculture River stretches in West Bengal
countries

40 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008

 AFRICAN CATFISH TRADITIONAL RIVER FISHING
IN INDIA

of the fish through various human activities in the river has
contributed to the population decline of the freshwater fish. The
recent invasion of alien species in riverine stretches has further
threatened the existence of local and endemic fishes (Singh and
Lakra, 2006).
The eroding health of the riverine ecosystem due to the invasion
of alien fish species is attributable to the following facts:
1 The ability of many introduced fish species to thrive in degraded
aquatic habitats and their potential to impact on aquatic ecosystem
structures and functions. Thus, introduced fish may represent
both a symptom and a cause of decline in river health and the
integrity of native aquatic communities.
2 The varying sensitivities of many commonly introduced fish
species (Cyprinus carpio, tilapia and Aristichthys nobilis, Clarias
gariepinus and Pangasius sutchi) to degraded stream conditions
to become invasive because of the environmental and biological
characteristics of the receiving water body, are all confounding
factors that may obscure the patterns of introduced fish species
distribution and abundance and therefore their reliability as
indicators of river health.
3 The relationships of alien species distributions and indices
of abundance and biomass with the natural environmental
features, the biotic characteristics of the local native fish
assemblages and indicators of anthropogenic disturbance
are subject to varying intensities of human interventions on
alien species introduction for aquaculture without adopting
biosecurity and bio-safety measures.
OREOCHROMIS NILOTICUS
FROM YAMUNA RIVER
4 Alien fish species found to be widespread in distribution and
often abundance in rivers and streams could be considered to
be relatively tolerant to river degradation, making them good
candidate indicators of river health.
5 Potential confounding factors that may influence the likelihood of
successful establishment of an alien species have the potential to
disrupt the biological integrity of natural aquatic ecosystems.
It is therefore important to assess the associated risks and benefits
of any alien fish introduction. If found appropriate, develop and
implement a plan for their responsible use. The development of
codes of practice is the mechanism to assist in the responsible use of
introduced species.
In India, the National Bureau of Fish Genetic Resources has also
developed plans and guidelines to regulate the introduction of exotic
fish to safeguard fish biodiversity and biological integrity. The

TABLE 2: Environmental impact of alien fish introductions in India.

FISH SPECIES ENVIRONMENTAL IMPACT REFERENCES

Oreochromis mossambicus
Aristichthys nobilis
Cyprinus carpio
O. niloticus
Displaced Gangatic carps, Puntius dubius and Labeo kontius and Singh and Lakra, 2006
now posing threat to Etroplus suratensis in back waters of Kerala
Displacement of Catla and silver carp, hybridization with silver Singh and Ponniah 2001; Mia et al 2005;Taylor 2005;
carp and Catla Singh and Lakra, 2006
Displacement of local spp. Schizothorax, Osteobrama belangiri, Tor Singh and Das 2006; Singh and Lakra 2006
putitora etc.
Reduced catches of indigenous fish species Sugunan 2002 ; Singh and Lakra 2006

Clarias gariepinus Environmental problem posing threat to biodiversity. Thakur 1998; Singh and Ponniah 2001 Singh and
Risks of hybridisation with native fishes, loss to local culturable fishes. Mishra 2001; Sugunan 2002; Singh and Lakra 2006
Hypophthalmichthys molitrix Naturalised in some reservoirs and displacement of Catla Pandey 1997; Singh 2004

Pangasius sutchi Evironmental problems are realised but yet to be documented. Under study by NBFGR

JUNE 2008 AQUACULTURE HEALTH INTERNATIONAL 41

RESEARCH FOCUS
SPREAD AND COLONIZATION OF ALIEN FISH SPECIES IN OPEN WATERS
HARVEST OF PANGASIUS SUTCHI
aquaculturists and farmers are advised to comply with the available
regulatory mechanisms for alien fish introductions, along with strict
conditions of sanitary and hygienic standards.
REFERENCES
Arthur JR 2005. An historical overview of pathogen introductions
and their trans-boundary spread in Asia. Preparedness and response
to aquatic animal heath emergencies in Asia. FAO Fisheries Technical
paper 451, FAO Rome, Italy. pp1-71
DIAS 2004. Database on Introductions of Aquatic Species. Fisheries
Global Information System. FAO Rome, Italy
IUCN/SSC Invasive species Specialists group 2000. Guidelines for
the prevention of biodiversity loss caused by alien invasive species.
IUCN Gland, Switzerland
Mabuchi K, Senou H and Nishida M 2008. Mitochondrial DNA
analysis reveals cryptic large-scale invasion of non-native genotypes
of common carp (Cyprinus carpio) in Japan. Molecular Ecology 17.
pp796-809
Mia M, Younus, Taggart JB, Gilmour AE, Gheyas AA, Das TK, Kohinoor
AHM, Rahman MA, Sattar MA, Hussain MG, Mazid MA, Penman D
OIE PUBLICATIONS 2008
Available for purchase from the Office International des
Épizooties (OIE), 7 Rue de Prony, 75017 Paris, France, or for
consultation on-line at www.oie.int
• Aquatic Animal Health Code, 11th edition 2008. English
language version. 170pp
• Code sanitaire pour les animaux aquatiques, 11th edition
2008. French language version. pp160
• Código Sanitario para los Animales Acuáticos, 11th edition
2008. Spanish language version. pp160
• Manual of Diagnostic Tests for Aquatic Animals, 5th edition
2006. English language version. pp469
• Manual de Pruebas de Diagnóstico para los Animales
Acuáticos. 5th edition 2008. Spanish language version.
pp480
• Changing trends in managing aquatic animal emergencies
(ed) Eva-Maria Bernoth. Scientific and Technical Review
27 (1): pp300 (trilingual English-French-Spanish). April
2008. (Editor’s note: This publication will be reviewed in a
forthcoming issue of Aquaculture Health International.)
• Risk analysis in aquatic animal health (ed) CJ Rodgers.
Thematic Publications 2001. pp346
42 AQUACULTURE HEALTH INTERNATIONAL JUNE 2008
COMMON CARP
and McAndrew BJ 2005. Detection of hybridisation between Chinese
carp species (Hypophthalmichthys molitrix and Aristichthys nobilis)
in hatchery brood stock in Bangladesh, using DNA microsatellite
loci. Aquaculture 247. pp267-273
Pandey AC 1997. Incidence and impact of capture of exotic carps in
the River Gomti near Gohania village (Distt. Sultanpur). Ind Journ
Anim Science 67. pp637-638
Sena De Silva, Thuy TT, Nguyen Nigel, Abery W and Amerasinghe US
2006. An evaluation of the role and impacts of alien finfish in Asian
Inland aquaculture. Aquaculture Research 37. pp1-17
Senanan W, Kapuscinki Anne, Uthairat Na-Nakorn R and Miller
LM 2004. Genetic impacts of hybrid catfish farming (Clarias
macrocephalus x Clarias gariepinus) on native catfish populations in
Central Thailand. Aquaculture 235. pp167-184
Singh AK 2004. Aquaculture diversification and species enhancement:
problems and perspectives. In: Zoology and Human Welfare (ed) Dr
Ashok Verma. Published by Dr SP Mukherji Government College,
University of Allahabad. pp252-261
Singh AK and Das P 2006. Status of common carp (Cyprinus carpio
– Linnaeus, 1758) in aquaculture and its environmental impact.
Aquaculture 7. pp245-257
Singh AK and Mishra A 2001. Environmental issue of exotic catfish
culture in Uttar Pradesh. J Envirn Biol 22. pp205-208
Singh AK and Ponniah AG 2001. In: Proceedings of the national
symposium on fish health management and sustainable aquaculture.
College of Fisheries, GB Pant University of Agriculture and
Technology, Nov 1-2, 2000 Pantnagar. (eds) Singh UP, Chauhan RS
and Sharma AP. pp93-96
Sugunan VV 2002. Clarias gariepinus (African catfish) gravitates
into the River Yamuna, Sutlej, Godawari. Angst comes true. Fishing
Chimes 22. pp50-52
Taylor RM and Pegg MA 2005. Management and ecological note
response of bighead carp to a bio-acoustic behavioural fish guidance
system. Fisheries Management & Ecology 12. pp283-286
Thakur NK 1998. A biological profile of African catfish, Clarias
gariepinus and the impact of its introduction into Asia. In: Fish
Genetics and Biodiversity conservation (eds) Ponniah AG, Das P, and
Verma SR. Natcon Publication, Muzaffarnagar (UP), India. pp275-292
CONTACT DETAILS
AK Singh and WS Lakra. National Bureau of Fish Genetic Resources,
Canal Ring Road, PO Dilkusha, Lucknow-226002 (UP), India. Email
aksingh56@rediffmail.com
Arvind Mishra. UP State Fisheries Department, Varanasi-221002,
Uttar Pradesh, India ■
Aquaculture without Frontiers (AwF)
is an independent non-profit organisation that assists in the alleviation of poverty
in developing countries by supporting projects designed to provide fish for food
and income through sustainable small-scale aquaculture. AwF has also assisted in
tsunami relief work.

So far we have project activities in Bangladesh , India , Indonesia , Malawi , Nepal

and Thailand and our AwF Volunteers have provided assistance in several other
countries including Ghana , Kenya , Liberia , Papua New Guinea and Peru .

Please help us to help others by donating yourself

or by organising fund-raising activities!

Further information on our activities can be found at:

www.aquaculturewithoutfrontiers.org

Aquaculture without Frontiers - be a part of something special.