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The Use of Probiotics in Shrimp Aquaculture

The Use of Probiotics in Shrimp Aquaculture

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MINIREVIEW
Theuseofprobioticsinshrimpaquaculture
Ali Farzanfar
Iranian Fisheries Research Organization, Tehran, Iran
Correspondence:
Ali Farzanfar, IranianFisheries Research Organization (IFRO), No.297, West Fatemi Ave., Tehran, Iran. Tel.:
1
98912 3153788; fax:
1
98 192 4562534; e-mail:afarzanfar@yahoo.comReceived 13 February 2006;accepted 7 April 2006.First published online 20 June 2006.DOI:10.1111/j.1574-695X.2006.00116.xEditor: Willem van Leeuwen
Keywords
shrimp; aquaculture; probiotic; lactic acidbacteria;
Streptococcus 
spp.;
Lactobacillus 
spp.;
Bacillus 
spp.
Abstract
Shrimp aquaculture, as well as other industries, constantly requires new techniquesin order to increase production yield. Modern technologies and other sciencessuch as biotechnology and microbiology are important tools that could lead to ahigher quality and greater quantity of products. Feeding and new practices infarming usually play an important role in aquaculture, and the addition of variousadditives to a balanced feed formula to achieve better growth is a common practiceof many fish and shrimp feed manufacturers and farmers. Probiotics, as ‘bio-friendly agents’such as lactic acid bacteria and
Bacillus
spp., can be introduced intothe culture environment to control and compete with pathogenic bacteria as wellas to promote the growth of the cultured organisms. In addition, probiotics arenonpathogenic and nontoxic microorganisms without undesirable side-effectswhen administered to aquatic organisms. These strains of bacteria have many other positive effects, which are described in this article.
Introduction
The use of probiotics as farm animal feed supplements datesback to the 1970s. They were originally incorporated intofeed to increase the animal’s growth and improve its healthby increasing its resistance to disease. The results obtained inmany countries have indicated that some of the bacteriaused in probiotics (Lactobacilli) are capable of stimulatingthe immune system (Fuller, 1992).The beneficial effect of the application of certain bene-ficial bacteria in human, pig, cattle and poultry nutritionhas been well documented. However, the use of suchprobiotics in aquaculture is a relatively new concept.With interest in treatments with friendly bacterial candi-dates increasing rapidly in aquaculture, several researchprojects that deal with the growth and survival of fishlarvae, crustaceans and oysters have been undertaken (Ali,2000).Yasudo and Taga (1980) predicted that some bacteriawould be found to be useful not only as food but also asbiological controllers of fish disease and activators of nutrient regeneration. It was only in the late 1980s that thefirst publication on biological control in aquacultureemerged, and since then the research effort has continually increased (Verschuere
et al 
., 2000).
Background of study
On fishes
Bacteria live in every corner of the aquatic environment. Thefish egg is the first stage of a fish life-cycle that could beexposed to bacteria. Therefore, a relatively dense, nonpatho-genic, and diverse adherent microbiota present on the eggswould probably be an effective barrier against the formationof a colony by pathogens on fish eggs. In addition, theestablishment of a normal gut microbiota may be regardedas complementary to the establishment of the digestivesystem, and under normal conditions it serves as a barrieragainst invading pathogens. Larvae may ingest substantialamounts of bacteria. It is obvious that the egg microbiotawill affect the primary colonization of the fish larvae(Verschuere
et al 
., 2000).Kennedy 
et al 
. (1998) used probiotic bacteria in theculture of marine fish larvae. They identified and usedprobionts for the culture of common snook, red drum,spotted sea trout and striped mullet. They then observedthat the application of probiotic bacteria to larval fish tanks(from egg through transformation) increased survival, sizeuniformity, and growth rate. The periodic addition of bacteria to the tanks altered the microbial communities of 
FEMS Immunol Med Microbiol
48
(2006) 149–158
c
2006 Federation of European Microbiological SocietiesPublished by Blackwell Publishing Ltd. All rights reserved
 
both tanks and fish. In addition, they noticed that the fisheggs incubated with probiotic bacteria were less likely todevelop bacterial overgrowth and die than those incubatedwithout probiotic bacteria.Carnevali
et al 
. (2004) isolated
Lactobacillus fructivorans
(AS17B) from sea bream (
Sparus aurata
) gut, and thenadministered it during sea bream development using
Bra-chinons plicatilis
and/or
Artemia salina
and dry feed asvectors. At the end of the experiments, they found asignificantly decreased larvae and fry mortality in theirtreated groups.Previously, Gildberg
et al 
. (1997) had analysed the effectof a probiotic of lactic acid bacteria in the feed of Atlanticcod fry (
Gadus morha
) on growth and survival rates. In theirstudy, a dry feed containing lactic acid bacteria (
Carnobac-terium divergens
) that had been isolated from adult intes-tines was given to cod fry. After 3 weeks of feeding the fry were exposed to a virulent strain of 
Vibrio anguillarum
. Thenumber of deaths was recorded during a further 3 weeks of feeding with feed supplemented with lactic acid bacteria. Acertain improvement in disease resistance was obtained, andat the end of the experiment lactic acid bacteria dominatedthe intestinal flora in surviving fish given feed supplementedwith lactic acid bacteria.Lara-Flores
et al 
. (2003) used two probiotic bacteria andthe yeast,
Saccharomyces cerevisiae
as growth promoters inNile tilapia (
Oreochromis niloticus
) fry. The results of thisstudy indicated that the fry subjected to diets with aprobiotic supplement exhibited greater growth than thosefed with the control diet. In addition, they suggested that the yeast is an appropriate growth-stimulating additive in tilapiacultivation.
On crustaceans
During the last few decades, aquaculture has become theworld’s fastest growing food production sector, with cul-tured shrimp growing at an annual rate of 16.8%. Mean-while, according to a World Bank report, global lossesresulting from shrimp diseases are around 3 billion USdollars. The potential negative consequences of using anti-biotics in aquaculture, such as the development of drug-resistant bacteria and the reduced efficiency of antibioticresistant for human and animal diseases, have led to sugges-tions of the use of nonpathogenic bacteria as probioticcontrol agents (Vaseeharan & Ramasamy, 2003).Moriarty (1999) reported on his successful experiences of using probiotic bacteria instead of antibiotics to control
Luminus vibrios
in shrimp farms in Negros, Philipine. Theeffects of ozone and probiotics on the survival of black tigershrimp (
Penaeus monodon
) were recorded by Meunpol
et al 
.(2003). They investigated the effects of ozone with andwithout feeds supplemented with the probiotic
Bacillus
S11on bacterial (
Vibrio harveyi
) growth and shrimp (
P. mono-don
) survival. According to the results of their study,shrimp survival after probiotic treatment, coupled withozonation, increased significantly compared with controls.The antagonistic effect of 
Bacillus
against the pathogenic
Vibrios
was evaluated in black tiger shrimp (
P. monodon
),and it was suggested as an alternative treatment factorinstead of antibiotics in shrimp aquaculture (Vaseeharan & Ramasamy, 2003).In another experiment that was performed by Rengpipat
et al 
. (2003), the growth and resistance to
Vibrio
in black tiger shrimp (
P. monodon
) fed with a
Bacillus
probiotic(BS11) were studied. It was found that the growth andsurvival rates of shrimps fed on the probiotic supplementwere significantly greater than those of the controls. Somestrains of Gram-negative bacteria have been used as probio-tics in shrimps too. For instance, Alvandi
et al 
. (2004)isolated
Pseudomonas
sp. PM11 and
Vibrio fluvialis
PM17as candidate probions from the gut of farm-reared subadultshrimp and tested for their effect on the immunity indica-tors of black tiger shrimp. The results of the study suggestthat the criteria used for the selection of putative probioticstrains, such as predominant growth on primary isolationmedia, ability to produce extracellular enzymes and side-ropheros, did not bring about the desired effect
in vivo
andimprove the immune system in shrimp.Nogami and Maeda (1992) found that production of crab(
Portunus trituberculatus
) larvae increased following theaddition of bacterial strain PM-4 to their culture water. Heisolated PM-4 from a crustacean culturing pond andcultured it in large quantities to add daily to the water of crab larvae. When bacteria increased to more than a specificpopulation, the protozoan population grew rapidly andreduced the bacterial population.
On bivalve mollusks
The mass culture of scallops and oysters has been introducedin many countries. However, mass mortalities of larvae havefrequently occurred, limiting the success of the hatcheries.To prevent these mortalities, most farmers routinely useantibiotics. As mentioned above, antibiotics have limitedapplicability, because of the ability of a large variety of pathogens to develop multiple antibiotic resistance. Analternative method for controlling pathogenic bacterialstrains in bivalve farms may be the addition of pure cultureof natural bacteria isolates (probiotics), which have beenshown through experimentation to produce chemical sub-stances inhibitory to bacterial pathogens (Gildberg
et al 
.,1997; Riquelme
et al 
., 1997; Vaseeharan & Ramasamy, 2003).
 Alteromons haloplanktis
was isolated from the gonads of Chilean scallop (
 Argopecten purpuratus
) brood stock anddisplayed
in vitro
inhibitory activity against the known
FEMS Immunol Med Microbiol
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2006 Federation of European Microbiological SocietiesPublished by Blackwell Publishing Ltd. All rights reserved
150
A. Farzanfar
 
pathogens
Vibrio ordalii
,
V. parahaemolyticus, V. anguillar-um, V. alginolyticus
and
Aeromonas hydrophila
. In an experi-mental infection, the
A. haloplanktis
and a
Vibrio
strain 11(that showed
in vitro
inhibition effects on
V. anguillarum
)protected the scallop larvae against the
V. anguillarum
(Riquelme
et al 
., 1997; Verschuere
et al 
., 2000).Douillet & Langdon (1994) added a bacteria strain (CA2)as a food supplement to larval cultures of the oyster
Crassostrea gigas
. They found more growth in larvae thathad been treated by CA2 bacteria cells.
On water quality
There are no serious problems for water quality during theinitial stages of farming aquatic organisms, when thestocked organisms are small and their metabolism rate andamounts of supplementary feed are low. However, with theprogress of culture the organisms grow, leading to a rapidincrease in biomass, and water quality deteriorates, mainly as a result of the accumulation of metabolic waste of cultured organisms, decomposition of unutilized feed, anddecay of biotic materials (Prabhu
et al 
., 1999). At this time,the application of a group of beneficial microorganisms(such as
Lactobacillus, Bacillus, Nitrosomonas, Cellulomonas, Nitrobacter, Pseudomonas, Rhodoseudomonas, Nitrosomonas
and
Acinetobacter 
) would be very useful for controlling thepathogenic microorganisms and water quality (Prabhu
et al 
., 1999; Shariff 
et al 
., 2001; Irianto & Austin, 2002).By definition, bacteria added directly to pond water arenot probiotics, and should not be compared with livingmicroorganisms added to feed (Rengpipat
et al 
., 2003).Many workers have evaluated some specific microorganismsas biological improvers for water quality: Douilett (1998)used a probiotic additive consisting of a blend of bacteria ina liquid suspension in intensive production systems. Theprobiotic blend improved water quality in fish and crusta-cean cultures by reducing the concentration of organicmaterials (OM) and ammonia. This procedure was accom-plished by a series of enzymatic processes carried out insuccession by the various strains present in the probioticblend. The addition of this blend to culture systems reducedthe concentration of 
Vibrio
strains and thus controlleddiseases caused by 
Vibrio
strains. In addition,
Bacillus
spp.have been evaluated as probiotics, with uses including theimprovement of water quality by influencing the composi-tion of water-borne microbial populations and reducing thenumber of pathogens in the vicinity of the farm species.Thus, the Bacilli are thought to antagonize potential patho-gens in the aquatic environment (Irianto & Austin, 2002).Bacterial species belonging to the genera
Bacillus, Pseudo-monas, Nitrosomonas
,
Nitrobacter, Acinetobacter 
and
Cellu-lomonas
are known to help in the mineralization of organicwater and in reducing the accumulation of organic loads(Shariff 
et al 
., 2001). Furthermore, there are many reports of the use of microbial products in aquaculture ponds forincreasing the removal rate of ammonia. Prabhu
et al 
.(1999) used some microorganisms in a shrimp farm toevaluate them as a factor for controlling the water quality.According to the results of this study, all factors of water-quality parameters were at optimum levels in the experi-mental ponds compared with the control.
On human consumption
The use of live microorganisms to enhance human health isnot new. For thousands of years, long before the discovery of antibiotics, people have been consuming live microbial foodsupplements such as fermented milks. According to Ayurve-da, one of the oldest medical sciences that dates back toaround 2500 BC, the consumption of yoghurt is recom-mended for the maintenance of overall good health. Ascientific explanation of the beneficial effects of lactic acidbacteria present in fermented milk was first provided in1907 by the Nobel Prize-winning Russian physiologist EliMetchnikoff. In his fascinating treatise ‘The Prolongation of Life’, Metchnikoff states that, ‘The dependence of theintestinal microbes on the food makes it possible to adoptmeasures to modify the flora in our bodies and to replacethe harmful microbes by useful microbes’ (Talwalkar, 2003).He proposed that the acid-producing organisms in fermen-ted dairy products could prevent ‘fouling’ in the largeintestine and thus lead to a prolongation of the life span of the consumer (Heller, 2001). Probiotics have a great variety of effects on human health. Probiotic therapy could be usedfor applications such as: modulation of the intestinalmicrobial communities, immune modulation, controllingallergic diseases, treating diseases related to the gastrointest-inal tract such as inflammatory bowel disease, and control-ling colorectal cancer and constipation (Ouwehand
et al 
.,2002).
Literature review on probiotics
Definitions and history
The word ‘probiotics’ originates from the Greek word ‘forlife’, and is currently used to name bacteria associated withbeneficial effects for humans and animals. The definition of probiotics has, however, evolved over time. Lily & Stillwell(1965) had originally proposed to use the term to describecompounds produced by one protozoan that stimulated thegrowth of another. The scope of this definition was furtherexpanded by Sperti in the early 1970s to include tissueextracts that stimulated microbial growth (Gomes & Mal-cata, 1999). Thereafter, other scientists applied the term toanimal feed supplements having a beneficial effect on the
FEMS Immunol Med Microbiol
48
(2006) 149–158
c
2006 Federation of European Microbiological SocietiesPublished by Blackwell Publishing Ltd. All rights reserved
151
Use of probiotics in shrimp aquaculture

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