1. March | April 2010
Feature title: Evaluation of probiotic bacteria in prawn production
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2. F: Probiotic
How should probiotics
be administered?
Probiotics could be used to replace
chemicals and antibiotics and have
already been shown to increase the
immune response, survival and disease
resistance in prawns (6). Probiotics
could either be incorporated into the
feed or administered into the rearing
water. Hai et al. (4) found that both feed
and water administered application was
efficient in the transfer of probiotics
to the prawns. However, survival, spe-
cific growth rates and total haemocyte
counts (THCs) concentrations were
higher when probiotics were supple-
mented through feed (4) .
Thus, feed incorporated probiotics are
more beneficial than probiotics adminis-
tered to the rearing water. THCs play
important roles in the crustacean immune
Evaluation of probiotic response as the haemocytes encapsulate
bacteria, thus a high haemocyte count clears
the bacterial load in the haemolymph.
or on the sediments. The spores of Bacillus
sp. can be easily introduced into dry food
Bacillus spp. to inhibiting the growth of
pathogenic bacteria could be one of the
bacteria in prawn production
and are unlikely to use genes for antibiotic enzymes produced by Bacillus, including
How do probiotics work? resistance or virulence from the vibrios or protease, amylase, lipase and gelatinase (6).
Proposed mode of actions of probiotics related gram-negative bacteria (6) and have The success of Bacillus spp. has been
includes immunostimulation, inhibition or not been associated with aquatic organism demonstrated by the development and use
by Tim O’brine, university of Plymouth, united Kingdom competition (antagonistic) with potential pathologies.
pathogens or simply maintenance of healthy For this
and diverse intestinal microbiota. reason, their
ECOBIOL
This article focuses on our current knowledge of the benefits of probiotic application, selecting probiotics Non pathogenic strains of Vibrio, use has been ®
and their modes of actions in combating microbial diseases in prawn production Aeromonas, Pseudomonas, Bacillus and promoted and
Alteromonas that are known to have antago- widely accept-
nistic properties to pathogenic strains ed within
can be used as probiotic strains (8). The the industry.
The probiotic for healthy growth
G
inhibitory compounds produced from the Banerjee et al.
lobally, two million metric and in the live feed and they usually cause of probiotics to manipulate the microbial antagonistic organisms can control the (9) observed 90
tonnes of prawn valued disease under sub-optimal culture condi- balance (3). pathogenic bacteria population in a system percent inhibi-
at US$14.4 billion were tions. Therefore the successful completion Probiotics are organisms that create without affecting the water quality and tion of patho-
produced in 2004(1), with of the larval cycle in prawns requires a a favourable environment for the growth therefore could be very beneficial to prawn genic bacteria
large industry growth observed in healthy microbial balance in the system (3). and well-being of the host animals by their production. by Bacillus
Asian countries. During the larval stages of prawn transience through the gastrointestinal tract Examples of antagonistic bacteria that subtilis along
production, luminescent vibriosis caused or merely by their presence in the water. secrete inhibitory substances include nisin with competi-
Intensification of prawn production has by Vibrio spp. is particularly severe and The selection of probiotics for specific (antibacterial peptide) of Lactobacillus lactis, tive exclusion
resulted in an increase of bacterial infec- sporadically causes temporary closure of marine aquatic species has to be considered pediocin (proteinaceous bacteriocin) of of patho-
tions which frequently cause mass mortali- economic activities in prawn larviculture carefully in order to make them spe- Pediococcus acidlactici, sidephore-like mol- genic vibrios
ties of prawn larvae due to the proliferation facilities (3). cies specific (4). Probiotic candidates are ecules and pyocyanin (antibiotic pigment) by Bacillus spp.
of opportunistic pathogenic bacteria. Vibriosis elicits signs of lethargy, degener- most often bacteria with inhibitory activity of Pseudomonas spp. The primary require- Moriar ty(6)
Disease is often the result of stress ation of the gills, lymphoid organs, digestive against a target pathogen or the dominant ment for the demonstration of antagonistic reported that, Improvements in growth and Feed Conversion Ratio
mode of action of probiotics in aquaculture Lower mortality in the ponds
caused by poor water quality and by reduc- glands and detachment of the epithelium intestinal bacterial flora in larvae from well when Bacillus
Increases biomass production
ing stress, less mortality may occur. lining the mid-gut, all of which lead to even- performing prawn-rearing groups. systems is the assurance of the minimum sp. was added Keeps pathogenic bacteria populations under control
The single most critical factor governing tual or rapid mortality. For example, Hill et al. (5) identified inhibitory concentration of the antagonistic to pond water Helps maintain optimal health status
optimal growth and survival of prawn is Application of disinfectants and antimi- Bacillus pumilis in the mid gut of P. monodon compound in the culture, which is supple- in penaeid
water quality. Prawns are delicate creatures crobials can complete the larval cycle in which had an inhibitory effect against patho- mented to the larval rearing tank (3). prawn culture,
and can be stressed by adverse environ- prawns but has led to the emergence of genic bacteria. The dominant bacterial flora Antagonistic, gram-positive Bacillus spp. there was a
mental conditions in culture ponds, result- antibiotic-resistant pathogens with threat is possibly capable of excluding invading have been used as a probiotic in fish and decrease in
ing in reduced appetite, slow growth, as well of gene transfer to humans as well as waste bacteria from the adhesion sites of the gut shellfish (9) and could be used in prawn luminous Vibrio rl
Natuna trition
as being more susceptible to disease (2). antimicrobials ending up in the environment. wall thus supporting the endemic bacterial aquaculture. densities.
u
Vibrio spp. are normally present in the These negative consequences of antibiotic flora of prawn - a more natural approach to Bacillus spp. are naturally ingested by The antago- NOREL,S.A. • Jesús Aprendiz, 19, 1º A y B • 28007 Madrid (SPAIN)
Tel. +34 91 501 40 41 • Fax +34 91 501 46 44 • www.norel.es
culture facilities, larval gut flora of prawns treatments have led to the development protecting prawns from diseases. prawns and other organisms that feed in nistic factor of
18 || InternatIonal AquAFeed || March-april 2010
18 InternatIonal AquAFeed March-april 2010 March-april 2010 || InternatIonal AquAFeed || 19
March-april 2010 InternatIonal AquAFeed 19

3. F: Probiotic
also reported that B. coagulans
used as a water additive at a
certain concentration could sig-
nificantly increase the survival
rate and some digestive enzyme
activities of prawn larvae. In
addition, the use of B. subtilis
was improved prawn immune
responses by increases in phe-
noloxidase activity, phagocytic
activity and clearance efficiency
against Vibrio alginolyticus.
Several probiotics
versus one probiotic
Combining probiotics to pro-
duce a range of beneficial effects
on prawn production has been
studied and is often employed
in commercial probiotics, as
previously mentioned by the
use of multiple Bacillus spp. in
SANOLIFE® MIC (10).
The combination of two pro-
biotics in Western king prawn
production was not found to
significantly improve the specific
growth rate of prawns compared
to single use of a probiotic,
but the prawns exposed to the
combined probiotics did elicit a
lower proportion of hyalinocytes
(involved in phagocytosis) as well
as having a lower bacterial load
in the haemolymph, hence were
of SANOLIFE® MIC, a mixture of strains super oxide radicals, resulting in the inhibi- healthier than those exposed to the
of Bacillus spp. Decamp et al. (10) reported tion of bacteria due to oxidative damage individual probiotics (4).
on the performance of SANOLIFE® MIC and lipid peroxidation. Far et al. (15) also compared the use of
on Asian and Latin-American hatcheries, As a result, prawns fed the probiotics one probiotic against multiple probiotics
with P. monodon and L. vannamei and found Pseudomonas synxantha and P. aeruginosa through feed incorporation. Treatment
that SANOLIFE® MIC performed as well as had higher specific growth rates, survival groups consisted of prawns fed diet
prophylactic use of antibiotics and was also and food conversion ratio compared to with B. subtilis and prawns fed a diet
a cost effective alternative. prawns not fed probiotics (11). Probiotics mixed with B. subtilis and a commercial
Pai et al. (3) suggested another mode of stimulated the prawns to produce a probiotic (BIO-GOLD, Korea) which con-
action of Bacillus; immunostimulation and higher THC and resulted in the lowest sisted of L. acidophilus, Bacillus subtilis
enhanced food acceptance which results in bacterial load in the haemolmpyh, thus and L. plantarum. Interestingly, diets just
improved general health of prawns rather improving the health of the prawns (4). consisting of B. subtilis had significantly
Dry with 20-50% less energy
than the antagonistic mode described above. Nakayama et al. (12) demonstrated higher survival and yield compared with
In their study they found that the bacteria that B. Subtilis could be effective in the diet incorporating both B. subtilis
that showed the greatest antagonism was the inhibition of Vibrio growth, while and BIO-GOLD. Thus it appears that B.
the Pseudomonas bacteria. B. Licheniformis and B. Megaterium subtilis alone, out performs BIO-GOLD,
The Pseudomonas bacteria suppressed could be effective in reducing V. harveyi but it should also be noted that a diet just
the vibrio population in a P. monodon larval haemolytic activities, suggesting that containing BIO-GOLD was not different
rearing system by production of pyocyanin compounds produced by Bacillus might from the diet containing B. Subtilis and
giving them a competitive advantage over be effective against Vibrio growth and BIO-GOLD.
V. harveyi, mediated by the killing of the toxin production. Therefore, an inhibitory action between
pathogen (3). Balcazar and Rohas-Luna (13)reported the consortium of probiotics in the BIO-
The unusual redox properties of pyo- that B. subtilis produced antimicrobial GOLD and B. subtilis diet can be ruled out.
cyanin and other phenazine compounds activity against pathogenic Vibrio spp. and Overall though, the addition of B. subtilis
secreted by fluorescent Pseudomonads can that prawn mortality was reduced by was found to improve prawn (L. vannamei) Geelen Counterflow / T +31-475-592315
enhance the production of hydroxide and treatment with B. subtilis. Zhou et al. (14) survival rate and yield (15). Geelen Counterflow USA Inc. / T +1-772-559-4338
Geelen Counterflow América Latina / T +54-9-2362-418899
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