July | August 2010

Feature title: Evaluation of Vibrio control with a multi-species

probiotic in shrimp aquaculture
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The International magazine for the aquaculture feed industry

 F: Vibro control

High end
machinery
for top quality output
Key machinery and
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for the aquatic feed industry
Evaluation of Vibrio control PELLET MILL
• Low energy consumption
with a multi-species probiotic in shrimp aquaculture • Low maintenance costs and excellent
durability
by Elisabeth Mayer MSc, Biomin, Austria
• Largest effective die surface
In the economically important panaeid shrimp, members of the microorganism genus Vibrio have become
• High lifetime of die and rollers
a major constraint on production and trade during the past two decades. They are responsible for several
diseases and mortalities of up to 100 percent, causing global losses of around US$3 billion. Shrimp disease
prevention and control are now priority research topics. In this article Elisabeth Mayer reports on in vivo
trials using AquaStar® as a probiotic feed additive in P. vannamei.

P
enaeid shrimp culture both in the hatchery and grow-out ponds, the normal microflora of farmed and wild
has become an important reduced feed conversion and growth rates panaeid shrimp. They become opportunistic
economic activity in many in surviving individuals, thus having a nega- pathogens when the natural defence mech-
countries, particularly tive impact on the overall financial efficiency anisms are suppressed (Lightner, 1993). In
in Asia and South America, where of the business. intensive systems, shellfish species are often
shrimp farming represents a substan- exposed to stressful conditions due to the
tial source of revenue. Vibriosis in shrimp high stocking density, leading to secondary
aquaculture vibriosis.
Shrimp accounts for about 20 percent of Vibriosis is a bacterial disease caused by Vibrio harveyi, a luminous marine bac-
the value of exported fishery products over gram-negative, motile, facultative anaerobe terium, is one of the most important
the past 20 years. bacteria of the family Vibrionaceae. It is etiological agents of mass mortalities of
ANDRITZ FEED & BIOFUEL extruders provide the new-
However, the shrimp farming industry ubiquitous throughout the world and all black tiger shrimp (Penaeus monodon) larval
est technology for production of the most demanding
is constantly under threat due to the marine crustaceans, including shrimp, are rearing systems. Epizootics occur in all life extrudates in the aquatic feed industries.
outbreak of infectious diseases and environ- susceptible. Vibrio species are the eminent stages but are more common in hatcheries. ANDRITZ is one of the global market leaders for supply-
mental problems. microorganisms in the marine environ- Vibrio parahaemolyticus is a halo- ing plants, equipment, and services for the animal feed in-
Bacteria are among the groups of ment and usually constitute the majority in philic bacterium distributed in temper- dustry (pet food industry, fish and shrimp feed industries).
microorganisms causing ate and tropical coastal With the efficient and simple ANDRITZ FEED & BIOFUEL
serious losses in shrimp waters throughout the “one-stop-shop” concept, we cover all operations in the pro-
Table 1: Enumeration of total Vibrio spp. and Enterococcus in shrimp
culture throughout the digestive tract after feeding with test diet for 6 weeks (by FISH technique) world (DePaola et al., duction flow. Our solutions are tailor-made to the specific re-
world. Members of the 2000). Some strains can quirements, be it for know-how, engineering, process lines,
CFU/g single unit machines, spare and wear parts, or service.
genus Vibrio, including cause acute gastroenteritis
V. parahaemolyticus and Hepatopancreas Intestine in humans, often after the
V. harveyi, have been Total Vibrio Entero-coccus Total Vibrio Entero-coccus consumption of contami-
described as the main (x 104) (x 106) (x 106) (x 108) nated seafood (Matsumoto
pathogenic species in et al., 2000).
shrimp and are responsi- Some Vibrio species
Control 68.8 +/- - 94.1 +/- -
ble for most of the larval 19.5a 68.2ns have very high growth
ANDRITZ FEED & BIOFUEL
deaths. rates under optimal con-
AquaStar® 1.7 +/- 56.5 +/- 29.5 +/- 7.8 +/- 5.7 Europe, Asia and South America:
These pathogens 0.9b 23.2 19.4 ditions. Disease transmis- andritz-fb@andritz.com
cause serious infections, sion can occur rapidly and USA and Canada:
a, b Means within a column with different superscripts differ significantly w w w. a a r s e n . co m andritz-fb.us@andritz.com www.andritz.com
decreased production (p < 0.05) is either via water or as

16 | InternatIonal AquAFeed | July-august 2010 July-august 2010 | InternatIonal AquAFeed | 17

F: Vibro control
Figure 1 and Figure 2: Survival and FCR of juvenile shrimp in biofloc culture system (p < 0.05)
a result of ingestion of infective
material, although there is some
evidence that wounds can also
provide a means of entry. The
pathogen releases exotoxins that
effectively break down the wall
of the gastrointestinal tract and
destroy the host´s immune cells.
Death can occur overnight after
acute outbreaks. (Peddie and
Picture 1: V. parahaemolyticus Wardle, 2005)
(Courtesy Dariano Krummenauer)
Signs of Vibrio disease
Vibrio infections are commonly
known as black shell disease, tail rot,
septic hepatopancreatic necrosis,
brown gill disease, swollen hindgut
syndrome and luminous bacterial
disease, describing a number of clini-
cal signs:
• Lethargy
• Loss of appetite
• Discoloured and necrotic
Picture 2: L. vannamei with greenish hepatopancreas with the pres-
fluorescence on the tail ence of ‘clumping’ (aggrega-
(Courtesy Dariano Krummenauer)
tion of digestive cells)
• Red discolouration of the body
• Yellowing of the gill tissue
• White patches in the
abdominal muscle
• Melanisation
• Granulomatous encapsula-
tion, necrosis and inflam-
mation of organs (lymphoid
organ, gills, heart etc.)
• Luminescence
Management strategies
for shrimp disease
prevention and control
Picture 3: Necrosis on the Use of antibiotics to control
muscular fiber caused by colonies these agents has led to problems
of V. parahaemolyticus
of drug resistance and resulted in
(Cortesy Dariano Krummenauer)
trade restrictions in export mar-
18 | InternatIonal AquAFeed | July-august 2010
kets. Shrimp aquaculture continues to find
more effective and environmentally friendly
approaches of improving shrimp health and
yields.
One such approach is the prevention
of infection by using specific pathogen free
(SPF) shrimp.
Such shrimp are genetically improved
stocks known to be free of one or more
specified pathogens and will ensure that
seed shrimp are not the conduit for intro-
duction of pathogens (Lotz, 1997).
However, SPF status is a temporary
condition which isn´t passed on genetically
and is lost once the SPF broodstock are
transferred to a commercial facility.
Vaccination or immunostimulation of
shrimp is another widely accepted technol-
ogy that promotes the immune response.
Since shrimp possess a non-specific immune
system without antibodies, they are not
enabled to specifically ‘remember’ expo-
sure to pathogens, which is the basis of
vaccination.
Consequently, the efficiency of response
on subsequent encounters may be limited.
Probiotics are another means of dis-
ease control which have found use in
aquaculture. The mode of action of the
probiotics is rarely investigated, but pos-
sibilities include competitive exclusion,
that is, the probiotics actively inhibit
the colonisation of potential pathogens
in the digestive tract by production of
bactericidal substances, competition for
nutrients and space, and modulation of
the immune-system. The stimulation of
host immunity and exclusion of patho-
gens may provide greater non-specific
disease protection as a result of both
immunity enhancement and competitive
exclusion (Rengpipat et al., 2000).
There is accumulating evidence that the
prophylactic use of beneficial bacteria is
Table 2: Mean survival, growth rate, final weight,
Reducing the
final biomass and FCR Vibrio load in the
intestine of shrimp
Control AquaStar®
A study by Dr Kidchakan
Supamattaya (2006) at the
Survival (%) 52.0a 83.0b Prince of Songkla University,
Growth rate (g/week) 0.85a 0.92b Thailand, has shown that the use
Final weight (g) 8.42a 9.05a
of AquaStar® Hatchery in feed
is effective in reducing the total
Final biomass (kg/tank) 45.97a 78.87b
number of Vibrio bacteria found
FCR 2.70a 1.40b in the hepatopancreas and intes-
Productivity (kg/m²) 1.31a 2.25b tine of white shrimp (Panaeus
a, b Means within a row with different superscripts differ vannamei), which can reduce the
significantly (p < 0.05); analyzed by one-way analysis of risk of infection (see Table 1).
variance (ANOVA). Groups of 20 juvenile white
shrimp (1 - 1.5g) were stocked
effective at inhibiting a wide range of fish into 200L glass aquaria and fed
pathogens. to satiation five times daily during a six-week
Recent data from in vivo experiments period. A commercial type diet was used
suggest that AquaStar® may be beneficial as a control. AquaStar® Hatchery was sup-
in the control of the Vibrio load in shrimp plemented to the feed at an inclusion rate
aquaculture. AquaStar® is a well-defined, of 0.5 percent. Six replicates for treatment
multi-strain probiotic product for fish were used. During the trial all data have been
and shrimp and promotes a beneficial recorded.
gut microflora as well as an improved Enterococcus faecium – as part of
environmental condition in shrimp and AquaStar® Hatchery - was found along the
fish ponds. shrimp digestive system in the group fed the
naturally supports…
… Per formance
… Health
… Stress management
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Tel +41 61 811 33 55 Fax +41 61 811 28 03
July-august 2010 | InternatIonal AquAFeed | 19
F: Vibro control
diet including this probiotic strain. The total
number of Vibrio spp. found in the hepatopan-
creas and intestine of shrimp fed AquaStar®
Hatchery was lower than control group.
In a recent study, the Marine Station of
Aquaculture at the Federal University of
Rio Grande (FURG) in Brazil, investigated
the effect of the simultaneous application of
AquaStar® Pond and AquaStar® Growout in
Litopenaeus vannamei cultured in a biofloc
technology system contaminated with Vibrio
parahaemolyticus (Krummenauer et al., 2009).
The juvenile white shrimp were stocked
in tanks at the high density of 300 shrimp/m².
Feed (38 percent crude protein) was
supplied three times/day. The experimental
group additionally received three grams of
AquaStar® Growout/kg feed and 0.5ppm/
week of AquaStar® Pond during the rear-
ing period. Biological parameters, growth,
weight gain, FCR and survival were evaluated
throughout the study for each group. The
experiment lasted for 70 days.
The results showed that AquaStar® was
effective in controlling Vibrio parahaemolyti-
cus in a biofloc culture system and improved
the overall productivity of the system. Survival
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 F: Vibro control Aqua News
Naturally ahead
Ghanaian aquaculture sector
receives
was increasedboost following
by 30 percent new Krummenauer
and FCR modernisation project by Fusion
D, Abreu PC, Lara G, Poersch L,
Encarnacao P, Wasielesky Jr W (2009) The Effect
Marine
parahaemolyticus and Emergence of Related Strains
Evidenced by Arbitrarily Primed PCR and toxRS

F
improved
usionsignificantly
Marine hasaswon well.anDespite the culture
phisticated systems.
of Probiotic along the
Fusion vannamei
in Litopenaeus coast
Biofloc Sequence Analyses. Journal of Clinical Microbiology
high density, the final weight of shrimp sup-
order to supply aquaculture marine will initially Technology
supplyCulture System contaminated
five poly- of the Gulf of Professor
38(2): Agius surveying the area last
578-585.
plemented with AquaStar
equipment
® was also slightly
for a new tilapia with Vibrio
ethylene circular parahaemolyticus.
fish pens Abstract World
but this Guinea. year to lay the foundations for this project
Peddie S, Wardle R (2005) Crustaceans: The impact
farm in Ghana in the first9.05g).
increased (8.42g versus mod- Thus, final is expected
number AquaculturetoConference,
increase Mexico.The new ini- and control of vibriosis in shrimp culture worldwide.

BIOMIN Aqua Products

biomass was
ernisation significantly
project of this higher
type forin the group
substantially once the farm tiative will initially
Lightner DV (1993) Diseases of cultured penaeid Aquaculture Health International, August: 4-5.
receiving ® resulting in a 70 percent
the West AquaStar
African country. becomes established.
shrimp. In: Mc-Vey JP (ed) CRCpurchase juvenile
hand book of
increase
Ghana inhas
production
excellent(79kg versus 46kg).
aquacul- The new farmmariculture,
will beCrustacean fish or2nd
locatedaquaculture, finger-
edn. CRC Rengpipat S, Rukpratanporn S, Piyatiratitivorakul S,
ture potential, but up until now downstream Press, fromBoca Raton, pp 393–486.
the Akosombo lings from com- Menasaveta P (2000) Immunity enhancement in black
Conclusion tiger shrimp (Penaeus monodon) by a probiont
Naturally ahead in aquaculture!
has relied on traditional and unso- dam on the River Volta. The water mercial
Lightner DV (1996) A Handbook of Shrimp hatch-
In order to withstand the high stocking
is of high quality and the water eries but within bacterium (Bacillus S11). Aquaculture 191: 271-28.
Pathology and Diagnostic Procedures for Disease
densities in shrimp production (hatcheries and
flow at the ofproposed Shrimp. 18
farm site
Cultured Penaeid World months it is
Aquaculture Supamattaya K , Viriyapongsutee B, Ruangsri J,
pond grow-out) and related stress situations,
is constant Society,
as it Baton
is controlled planned to have
Rouge, LA, p 4-1-4-27. Encarnacao P, Schatzmayr G (2005) Effect of
directly-fed probiotics are a promising additive
through the Lotz in house state-of-the-art hatchery
dam.JM (1997) Viruses, biosecurity mistic it willfaecium
probitoic Enterococcus contribute substantially to
and Phycophytic
and specific
to stimulate shrimp growth and secure The a lowfarm is planned for an area to meet the growing fingerling Substances onfish pen
Growth aquaculture
Performancedevelopments
and Health in
pathogen-free stocks in shrimp aquaculture. World
disease response. The data of these where studiesunemployment is high and requirements of the project. Condition
The of White Shrimp
Africa as (Penaeuscomes
this industry vannamei).
of age in
Journal of Microbiology and Biotechnology 13(4):
suggest
Simplethat the use offor
net enclosures AquaStar® improved
it is expected to create a sub- first phase is expected to be com- this vast continent. ”
405-413.
survival, growth
tilapia are rates,but
common and the generalstantial
health number of jobs. It is also pleted by September 2010, the
affordofpoor
status operation
juvenile controlvannamai while
Litopenaeus Lundin GG (1996) Fish health and quarantine. In:
within easy reach of the main new hatchery by middle of 2011, more inFormation:
also reducing pathogenic Vibrio spp. markets of the Global Attempts to Address Shrimp Disease. pp. 45. Related companies:
capital Accra. and further expansion of the pen
Marine/Environmental Paper No. 4. Land , Water and Sino-Aqua Fusion Marine Limited
Corporation
The project willhabitats
natural star t Division, systems by
with a Environment the end of that year.22F-2,NO.110
Department, The Marine
San-TuoResource
4th RoadCentre
Ling-Ya
References production of 200 metric
World Bank, Rome. tonnes Professor Carmelo Agius, Fusion
District, Barcaldine,
Kaohsiung 802Oban, Argyll
Taiwan
DePaola A, Nordstrom JL, Bowers JC, Wells of JG, tilapia per annum but the Marine international aquaculture Tel: 88 673PA37 1SE , Scotland
308868
Matsumoto C, Okuda J, Ishibashi M, Iwanaga Fax: 88 Tel:
673 + 44 1631 720730
3017389
David
MostWCof(2003) long-term plan
Seasonal abundance of total
the cages is to reach 2000 consultant said,M,
“Fusion Marine tech- Fax: + 44 1631 720731
Vi Garg P, Rammamurthy T, Wong H, DePaola Email: sales_dept@sino-aqua.com
and pathogenicused
currently Vibrio in
parahaemolyticus
Ghana tonnes per annum and then to nology has already proven itself
in Alabama in www.sino-aqua.com
Website: Email: enquiries@fusionmarine.com
oysters. Applied and Environmental Microbiology A, Kim YB, Albert MJ, Nishibuchi M (2000)
are very rudimentary diversify into marine species other parts of Africa and we are opti- Website: www.fusionmarine.com
69(3): 1521-1526. Pandemic Spread of an O3:K6 Clone of Vibrio
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