fry, quarantine measures, optimized feedingand good husbandry techniques. Monitoringdiseases by surveillance and reporting isessential, as is good sanitation and anemphasis on being responsible in the useof chemicals and antibiotics when diseasesoccur.There will certainly be a role for vac-cination as the overall emphasis changesto preventing diseases rather than treatingor curing them. Vaccination on its own isnot sufficient on its own to guarantee highsurvival and profitability (see box), but it isone of the tools to employ in good healthmanagement strategies.production, development and sustainableexpansion of marine fish aquaculture inthe Asian region. Most Asian aquacultureis operated on a small scale. Added to theother factors mentioned and the diversityof organisms in tropical waters, this contrib-utes to a truly challenging disease situationin which pathogens have various opportuni-ties for entry points. Only now are movesbeing made to correct the failure to collect,report and analyse epidemiological data inthe event of a disease outbreak.On a more positive note, more attentionhas been given in past few years to the iden-tification of aetiological agents involved infish disease epidemics. The following studiesgive a solid indication of the diversity of major pathogens affecting the fish farmingindustry in Asia (Bondad-Reantaso et al.,2005; Komar et al., 2005; Labrie et al., 2005;Leong et al., 2005, 2006 and Tan et al., 2003).Meanwhile, significant progress has beenmade in the field of vaccine research anddevelopment (Grisez and Tan, 2005).But as Asian aquaculture continues togrow, inevitably its problems with diseasewill worsen unless key steps are taken. Newdisease control concepts must be imple-mented. This includes the use of healthywere significantly lower than for all infectedgroups. Although not significantly differentfrom the 0.10 ml vaccinated fish for per-centage mortality, the 0.05 ml vaccine groupregistered a lower RPS of 45.76%.
Higher survival andlower mortality
Clearly, therefore, injecting 0.10 ml of the bivalent vaccine gave a higher rela-tive survival and the lowest mortality rateof any group in the test. The test alsodemonstrated that the European model of using vaccination as a primary prophylactictool could be applied successfully in Asia toreduce losses in spotted grouper juvenilesfrom a Vibrio infection.This points to a new direction in Asiangrouper culture for health management. Sofar, the development of effective strategies inAsia for disease prevention and control hasbeen hampered by a lack of data on the widevariety of disease-causing agents present andthe immune systems of Asian fish species.Technical support in such matters as diseasediagnosis is often lacking at farm level.There is also a stark contrast betweenthe Asian situation and modern salmonidaquaculture in countries such as Norway.Intensified salmon production has broughta separation between the hatchery sitesproducing fry and those for grow-out,together with optimized feed strategies andvirtually disease-free fingerlings. However,the segregation in year classes that isobligatory for salmon in Europe does notyet exist in Asia.Asian farms also commonly producedifferent species of fish at the same site,they make widespread use of trash fish asfeed and their fry are often wild-caught orderived from wild-caught brood stock. Withfarm licensing and zoning policies virtuallynon-existent in most Asian countries, thespread of diseases is promoted by havingtoo many fish and too many farms in aconcentrated area.It explains why the salmon farms of Norway or Chile consider asurvival rate of lower than95% as a sign of a diseaseoutbreak, whereas a survivalrate of 50% at fish farms inAsia is often considered tobe acceptable.
The biggestconstraint toprocution
Disease undoubtedlyrepresents one of the big-gest constraints to thelated by the Reed and Muench procedure asthe basis for deciding the optimal challengedose to be used during testing of vaccineefficacy.Figure 1 illustrates the clear doseresponse curve found from the virulenceassays. The LD100 dose (killing 100% of fish within three days) for this particularisolate of Vibrio cholerae was 1x1010 CFU/ml, with the LD50 established as 1x108CFU/ml.At 21 days after immunization, the vacci-nated and control fish were challenged withthe LD100 as a 0.10 ml injection adminis-tered intraperitoneally while the positivecontrols received 0.10 ml of sterile saline.Monitored for three days post-challenge,there was a 100% mortality rate for the fishin the positive control group. All dead fish inthe challenge experiment were confirmedby re-culture and re-identification using API20E as positive for Vibrio cholerae infection.Results of the efficacy test in Table 1confirmed that giving 0.10 ml of the vaccineprovided the highest rate of protectionagainst Vibrio cholerae. This vaccination gaverelative percentage survival (RPS) values of up to 77.59% compared with the relevantcontrols. Its percentage mortality rates0.9 % (v/v) saline. The aim was to findpathogen doses that would kill 50% and100% of injected fish within three days, sothat the mean lethal dose could be calcu-
Dr John S Clark
obtained his Ph.D.in micro-encapsulation at Heriot-WattUniversity in Edinburgh, Scotland in 1983.He currently works on formulation of anti-parasitics, vaccines, nutritional/health products for a variety of clients.Email: email@example.com
Dr Nantarika Chansue
is theAssociate Professor and Directorof the Veterinary Medicine AquaticAnimal Research Centre (VMAARC) atChulalongkorn University in Bangkok.She specializes in disease identifica-tion and control methods for shrimp,marine and freshwater fish, turtles andaquatic mammals. Aquatic immunoto-xicology has been her major researchinterest for some 20 years.
Professor Jirasak Tangtrongpiros
is the head of the Veterinary MedicineAquatic Animal Research Centre(VMAARC) at ChulalongkornUniversity in Bangkok, Thailand. Hespecialises in aquatic animal diseaseand his primary interest is in methodsfor disease prevention and control.
The authors would like to convey their thanks to all the staff of VMAARC inChulalongkorn University, BangkokThailand, and to the supporting staff of the Research Institute at Rayong, Rayong Province, Thailand, and finally to Drs. Gee,Na, Lin and Jim for their endless patience.References are available on request.
Measures to sustain asuccessful aquaculture
(Tan et al., 2006)
Use healthy (not necessarilycheap) fry
Quarantine incoming animals
Use formulated pelleted feed
Grade fish periodically
Monitor water quality
Record diseases and feeding
Observe withdrawal period of drugs
Remove dead fish at least once aday
Clean and disinfect equipment
Vaccination if available
Place your farm too close toothers
Have several species in one farm
Use fingerlings from unknownsources
Overstock (to overcome lowsurvival)
Use trash fish
Use drugs without diagnosis
Leave or throw dead fish in thewater
Restock fish without cleaning thecages
Ignore diseases until heavy mor-tality occurs
Percentage mortality of juvenile orange-spotted grouper,non-vaccinated and vaccinated by two intraperitoneally injecteddoses of a Vibrio vaccine and relative percentage survival (RPS)after challenge with Vibrio cholerae.GroupMortality*RPSNCControl, no infection2.63±3.72c97.44PCControl, normal saline IP100.00±0.00a0.00T10.05 ml. vaccine IP54.04±31.73b45.76T20.10 ml. vaccine IP23.15±18.61bc77.59
Data was subjected to two way analysis of variance using an SPSSprogram. Significant differences were determined at P<0.05.
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