evaluation of vibrio control with a multi-species probiotic in shrimp aquaculture
TRANSCRIPT
July | August 2010
Feature title: Evaluation of Vibrio control with a multi-species probiotic in shrimp aquaculture
The International magazine for the aquaculture feed industry
International Aquafeed is published five times a year by Perendale Publishers Ltd of the United Kingdom.All data is published in good faith, based on information received, and while every care is taken to prevent inaccuracies, the publishers accept no liability for any errors or omissions or for the consequences of action taken on the basis of information published. ©Copyright 2009 Perendale Publishers Ltd. All rights reserved. No part of this publication may be reproduced in any form or by any means without prior permission of the copyright owner. Printed by Perendale Publishers Ltd. ISSN: 1464-0058
In the economically important panaeid shrimp, members of the microorganism genus Vibrio have become 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.
Penaeid shrimp culture has become an important economic activity in many countries, particularly
in Asia and South America, where shrimp farming represents a substan-tial source of revenue.
Shrimp accounts for about 20 percent of the value of exported fishery products over the past 20 years.
However, the shrimp farming industry is constantly under threat due to the outbreak of infectious diseases and environ-mental problems.
Bacteria are among the groups of microorganisms causing serious losses in shrimp culture throughout the world. Members of the genus Vibrio, including V. parahaemolyticus and V. harveyi, have been described as the main pathogenic species in shrimp and are responsi-ble for most of the larval deaths.
These pathogens cause serious infections, decreased production
both in the hatchery and grow-out ponds, reduced feed conversion and growth rates in surviving individuals, thus having a nega-tive impact on the overall financial efficiency of the business.
Vibriosis in shrimp aquaculture
Vibriosis is a bacterial disease caused by gram-negative, motile, facultative anaerobe bacteria of the family Vibrionaceae. It is ubiquitous throughout the world and all marine crustaceans, including shrimp, are susceptible. Vibrio species are the eminent microorganisms in the marine environ-ment and usually constitute the majority in
the normal microflora of farmed and wild panaeid shrimp. They become opportunistic pathogens when the natural defence mech-anisms are suppressed (Lightner, 1993). In intensive systems, shellfish species are often exposed to stressful conditions due to the high stocking density, leading to secondary vibriosis.
Vibrio harveyi, a luminous marine bac-terium, is one of the most important etiological agents of mass mortalities of black tiger shrimp (Penaeus monodon) larval rearing systems. Epizootics occur in all life stages but are more common in hatcheries.
Vibrio parahaemolyticus is a halo-philic bacterium distributed in temper-
ate and tropical coastal waters throughout the world (DePaola et al., 2000). Some strains can cause acute gastroenteritis in humans, often after the consumption of contami-nated seafood (Matsumoto et al., 2000).
Some Vibrio species have very high growth rates under optimal con-ditions. Disease transmis-sion can occur rapidly and is either via water or as
by Elisabeth Mayer MSc, Biomin, Austria
Evaluation of Vibrio control with a multi-species probiotic in shrimp aquaculture
Table 1: Enumeration of total Vibrio spp. and Enterococcus in shrimp digestive tract after feeding with test diet for 6 weeks (by FISH technique)
CFU/g
Hepatopancreas Intestine
Total Vibrio(x 104)
Entero-coccus(x 106)
Total Vibrio(x 106)
Entero-coccus(x 108)
Control 68.8 +/- 19.5a
- 94.1 +/- 68.2ns
-
AquaStar® 1.7 +/- 0.9b
56.5 +/- 23.2
29.5 +/- 19.4
7.8 +/- 5.7
a, b Means within a column with different superscripts differ significantly (p < 0.05)
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F: Vibro control
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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
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 Wardle, 2005)
Signs of Vibrio diseaseVibrio 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 hepatopancreas with the pres-ence of ‘clumping’ (aggrega-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
Use of antibiotics to control these agents has led to problems of drug resistance and resulted in trade restrictions in export mar-
Figure 1 and Figure 2: Survival and FCR of juvenile shrimp in biofloc culture system (p < 0.05)
Picture 1: V. parahaemolyticus(Courtesy Dariano Krummenauer)
Picture 2: L. vannamei with greenish fluorescence on the tail(Courtesy Dariano Krummenauer)
Picture 3: Necrosis on the muscular fiber caused by colonies of V. parahaemolyticus (Cortesy Dariano Krummenauer)
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F: Vibro control
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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
Reducing the Vibrio load in the intestine of shrimp
A study by Dr Kidchakan Supamattaya (2006) at the Prince of Songkla University, Thailand, has shown that the use of AquaStar® Hatchery in feed is effective in reducing the total number of Vibrio bacteria found in the hepatopancreas and intes-tine of white shrimp (Panaeus vannamei), which can reduce the risk of infection (see Table 1).
Groups of 20 juvenile white shrimp (1 - 1.5g) were stocked into 200L glass aquaria and fed
to satiation five times daily during a six-week period. A commercial type diet was used as a control. AquaStar® Hatchery was sup-plemented to the feed at an inclusion rate of 0.5 percent. Six replicates for treatment were used. During the trial all data have been recorded.
Enterococcus faecium – as part of AquaStar® Hatchery - was found along the shrimp digestive system in the group fed the
effective at inhibiting a wide range of fish pathogens.
Recent data from in vivo experiments suggest that AquaStar® may be beneficial in the control of the Vibrio load in shrimp aquaculture. AquaStar® is a well-defined, multi-strain probiotic product for fish and shrimp and promotes a beneficial gut microflora as well as an improved environmental condition in shrimp and fish ponds.
Table 2: Mean survival, growth rate, final weight, final biomass and FCR
Control AquaStar®
Survival (%) 52.0a 83.0b
Growth rate (g/week) 0.85a 0.92b
Final weight (g) 8.42a 9.05a
Final biomass (kg/tank) 45.97a 78.87b
FCR 2.70a 1.40b
Productivity (kg/m²) 1.31a 2.25b
a, b Means within a row with different superscripts differ significantly (p < 0.05); analyzed by one-way analysis of
variance (ANOVA).
18 | InternatIonal AquAFeed | July-august 2010 July-august 2010 | InternatIonal AquAFeed | 19
F: Vibro control
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parahaemolyticus and Emergence of Related Strains Evidenced by Arbitrarily Primed PCR and toxRS Sequence Analyses. Journal of Clinical Microbiology 38(2): 578-585.
Peddie S, Wardle R (2005) Crustaceans: The impact and control of vibriosis in shrimp culture worldwide. Aquaculture Health International, August: 4-5.
Rengpipat S, Rukpratanporn S, Piyatiratitivorakul S, Menasaveta P (2000) Immunity enhancement in black
tiger shrimp (Penaeus monodon) by a probiont bacterium (Bacillus S11). Aquaculture 191: 271-28.
Supamattaya K , Viriyapongsutee B, Ruangsri J, Encarnacao P, Schatzmayr G (2005) Effect of probitoic Enterococcus faecium and Phycophytic Substances on Growth Performance and Health Condition of White Shrimp (Penaeus vannamei).
Krummenauer D, Abreu PC, Lara G, Poersch L, Encarnacao P, Wasielesky Jr W (2009) The Effect of Probiotic in Litopenaeus vannamei Biofloc Technology Culture System contaminated with Vibrio parahaemolyticus. Abstract World Aquaculture Conference, Mexico.
Lightner DV (1993) Diseases of cultured penaeid shrimp. In: Mc-Vey JP (ed) CRC hand book of mariculture, Crustacean aquaculture, 2nd edn. CRC Press, Boca Raton, pp 393–486.
Lightner DV (1996) A Handbook of Shrimp Pathology and Diagnostic Procedures for Disease of Cultured Penaeid Shrimp. World Aquaculture Society, Baton Rouge, LA, p 4-1-4-27.
Lotz JM (1997) Viruses, biosecurity and specific pathogen-free stocks in shrimp aquaculture. World Journal of Microbiology and Biotechnology 13(4): 405-413.
Lundin GG (1996) Fish health and quarantine. In: Global Attempts to Address Shrimp Disease. pp. 45. Marine/Environmental Paper No. 4. Land , Water and natural habitats Division, Environment Department, World Bank, Rome.
Matsumoto C, Okuda J, Ishibashi M, Iwanaga M, Vi Garg P, Rammamurthy T, Wong H, DePaola A, Kim YB, Albert MJ, Nishibuchi M (2000) Pandemic Spread of an O3:K6 Clone of Vibrio
was increased by 30 percent and FCR improved significantly as well. Despite the high density, the final weight of shrimp sup-plemented with AquaStar® was also slightly increased (8.42g versus 9.05g). Thus, final biomass was significantly higher in the group receiving AquaStar® resulting in a 70 percent increase in production (79kg versus 46kg).
ConclusionIn order to withstand the high stocking
densities in shrimp production (hatcheries and pond grow-out) and related stress situations, directly-fed probiotics are a promising additive to stimulate shrimp growth and secure a low disease response. The data of these studies suggest that the use of AquaStar® improved survival, growth rates, and the general health status of juvenile Litopenaeus vannamai while also reducing pathogenic Vibrio spp.
ReferencesDePaola A, Nordstrom JL, Bowers JC, Wells JG, David WC (2003) Seasonal abundance of total and pathogenic Vibrio parahaemolyticus in Alabama oysters. Applied and Environmental Microbiology 69(3): 1521-1526.
20 | InternatIonal AquAFeed | July-august 2010
F: Vibro control
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EDITOR’S DESK
WELC
OM
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INTE
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ATIO
NA
L AQ
UA
FEED
MA
GA
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E
Fusion Marine has won an order to supply aquaculture equipment for a new tilapia
farm in Ghana in the first mod-ernisation project of this type for the West African country.
Ghana has excellent aquacul-ture potential, but up until now has relied on traditional and unso-
phisticated culture systems. Fusion marine will initially supply five poly-ethylene circular fish pens but this number is expected to increase substantially once the farm becomes established.
The new farm will be located downstream from the Akosombo dam on the River Volta. The water is of high quality and the water flow at the proposed farm site is constant as it is controlled through the dam.
The farm is planned for an area where unemployment is high and it is expected to create a sub-stantial number of jobs. It is also within easy reach of the main markets of the capital Accra.
The project will star t with a production of 200 metric tonnes of tilapia per annum but the long-term plan is to reach 2000 tonnes per annum and then to diversify into marine species
along the coast of the Gulf of Guinea.
The new ini-tiative will initially purchase juvenile fish or finger-lings from com-mercial hatch-eries but within 18 months it is planned to have in house state-of-the-art hatchery to meet the growing fingerling requirements of the project. The first phase is expected to be com-pleted by September 2010, the new hatchery by middle of 2011, and further expansion of the pen systems by the end of that year.
Professor Carmelo Agius, Fusion Marine international aquaculture consultant said, “Fusion Marine tech-nology has already proven itself in other parts of Africa and we are opti-
mistic it will contribute substantially to fish pen aquaculture developments in Africa as this industry comes of age in this vast continent.”
more inFormation:Fusion Marine LimitedThe Marine Resource CentreBarcaldine, Oban, Argyll PA37 1SE , Scotland Tel: + 44 1631 720730Fax: + 44 1631 720731Email: [email protected]: www.fusionmarine.com
Ghanaian aquaculture sector receives boost following new modernisation project by Fusion Marine
Professor Agius surveying the area last year to lay the foundations for this project
Simple net enclosures for tilapia are common but afford poor operation control
Most of the cages currently used in Ghana are very rudimentary
2 | InternatIonal AquAFeed | July-august 2010 July-august 2010 | InternatIonal AquAFeed | 3
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VOLUME 13 I S SUE 4 2 010
Inclusion of a mineral premix for low-salinity culture
of Litopenaeus vannamei
Evaluation of Vibrio control
- with a multi-species probiotic in shrimp aquaculture
Recent advances in the use of diformates in fish
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