Probiotics and their applications in fish culture have been studied and discussed for much of the past decade. However, we must not continue to use the umbrella term too loosely and be mindful that not all probiotics display the same properties or yield the same subsequent benefits. Probiotics are inclusive of a wide range of microbes: Gram-positive bacteria, Gram-negative bacteria, yeasts and cyanobacteria have all shown some varying degree of success when applied to fish. They provide benefits to the host fish, in part at least, by improving the microbial balance and functionality of the host gastrointestinal microbial communities.

1. July | August 2010
Feature title: The Probiotic approach to elevating health of fish
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2. F: Probiotic F: Feature F: Vibro control
F: Probiotic
Table 3: Growth results during week 3 and 4 with lower feeding frequency
average weight total Solubility and digestibility lower salinity anymore, maybe because 10ppt and lower salinity, which confirms Reducing the OFIMER (Office National Interprofessionnel
diet including this probiotic strain. The total
Table 2: Mean survival, growth rate, final weight,
Growth
The lower leaching rate (higher reten- shrimp adapted to this condition. The best
final biomass and FCR
earlier results of leaching and digestibility.Vibrio load in the des Produits de laspp. found inl'Aquaculture),
number of Vibrio Mer et de the hepatopan-
salinity feed day 14 day 21 day 28 (g/week)
growth was obtained with diet number
tion) could be the result of a lower solubil- Based on these results a special mineral intestine of shrimp demonstrated thatof P. acidilactici could®
creas and intestine shrimp fed AquaStar
Control AquaStar®
The Probiotic approach
ity of the mineral; which could affect the three. premix was developed jointly by companies study by Dr Kidchakan improve rainbow trout resistance to yers-
A Hatchery was lower than control group.
digestibility of this minerals for the shrimp. For week five and six the feeding fre- Prayon and CreveTec. Supamattaya (2006) at the iniosis, a recent study, theby reduced accu-
In as demonstrated Marine Station of
10 ppt 1 10,05 11,68 12,82 1,39 quency was further lowered to only Survival (%)
Therefore, the apparent digestibility was two 52.0a 83.0b Prince of Songkla University, mulated mortalities after an intraperitoneal
Aquaculture at the Federal University of
tested for some selected growth trials. It times per day. Thailand, has shown that the use injection challenge.
Rio Grande (FURG) in Brazil, investigated
to elevating health of fish
Growth rate (g/week) 0.85a 0.92b
10 ppt 2 10,76 12,20 13,12 1,18 is clear from Table 1 that for all diets, the To understand the influence of leaching of AquaStar® Hatchery in feed the effect as benefits to intestinal morphol-
As well of the simultaneous application of
Final weight (g) 8.42a 9.05a
on growth, the feed for the three replicates
digestibility of selected minerals is not the AquaStar® Pond and AquaStar® Growout in
is effective in reducing the total ogy and disease resistance, P. acidilactici can
10 ppt 3 9,79 11,24 12,44 1,33
problem. in tank number one was Final biomass (kg/tank)more 45.97a
treated before inFormation:
78.87b
number of Vibrio bacteria found also alleviate vannamei trout vertebralbiofloc
Litopenaeus rainbow cultured in a col-
Figure 2. Probiotic P.
Molecular profiling of gastro-intestinal biota and effects on gut ultra-structural characteristicsfeeding: Four nets received normal feeds; FCRFrederic 2.70a
10 ppt 4 10,83 11,74 13,02 1,1 Martens
1.40b in the hepatopancreas and intes- umn compression syndrome (VCCS) Vibrio
acidilactici significantly
technology system contaminated with and
Material and methods four nets received feeds whichProductivity (kg/m²)Prayon SA, Rue J Wauters 144
were first 1.31a 2.25b tine of white shrimp (Panaeus improve aesthetic (Krummenauer et al., 2009).
elevated the circulating parahaemolyticus quality by lowering con-
4 ppt 1 9,78 11,05 11,97 1,1 From the leaching experiment, the most soaked in fresh water for onebhour before a row withEngis, Belgium
a, Means within 4480 different superscripts differ vannamei), which can reduce the dition factor (Aubin et shrimp were stocked
leucocyte levels and serum The juvenile white al. 2005a; Merrifield
by Dr Daniel L Merrifield and Matthew Emery, Senior microbiology technician,promising combinations were and Biologicalfeeding and four nets received feeds which
The School of Biomedical used for a Email: fmartens@prayon.be
significantly (p < 0.05); analyzed by one-way analysis of lysozymeof infection (see Table 1).
risk activity of tilapia et al. 2010a). These benefitsof 300 shrimp/m².
in tanks at the high density which improve
Sciences, The University of8,64
4 ppt 2 Plymouth, Plymouth, UK, Email: Daniel.merrifield@plymouth.ac.uk
10,06 11,38 1,37
feed trial. Four diets were produced on were first soaked in tank water (10ppt) for
Tel: +32 4 2739310
variance (ANOVA). after 14 days feeding white the quality(38 the final product protein) was
Website: www.prayon.com Groups of 20 juvenile Feed of percent crude could effec-
a small pellet mill with a diameter of two one hour before feeding (see table 4). (Ferguson et al. 2010)
4 ppt 3 9,18 10,64 12,16 1,99 shrimp (1 - 1.5g) were stocked tively boost production and elevate trout
supplied three times/day. The experimental
millimeters. There was a clear influence ofat inhibiting a wideDe Muylder fish
effective leaching Eric range of into 200L glass aquaria and fed farming additionally received three grams of
group profit margins.
4 ppt 4 10,00 11,50 12,34 1,17 Shrimp (Litopenaeus vannamei) were on growth, indicating thatpathogens. nutri- CreveTec
leaching of a foundation for enhanced
P
ments provide to satiation five times daily during a six-week
can infect and cause severe tissue damage to AquaStar® Growout/kg feed and 0.5ppm/
Email: eric@crevetec.be
robiotics and their applica- probiotic in fish culture has been under- imported from Thailand and of the to two ents is an imported factor Recent growth maintaining epithelial
microvilli morphology raised enterocytes in the data but period. A intestine of salmonid was used The financial ® Pond during the rear-
digestive function from in vivo experiments the anteriorcommercial type diet fish. From week of AquaStar benefits
10 ppt average 10,36 11,71 12,85 1,25 Website: www.crevetec.be
tions in fish culture have taken; information regarding the benefits of grams. They anterior intestine of rainbow trout shrimp. Leaching for integrity isinparamount ® may be beneficial these control. AquaStar® Hatchery was sup- of this probiotic parameters, growth,
in the were then transferred to the of one hourthat10ppt
suggest AquaStar to defend against as a findings we hypothesise that dietary ing period. Biological
4 ppt been studied and discussed
average 9,40 11,96 1,28 CreveTec-AFT Merrifield et al. 2010b).
10,81 P. acidilactici is available for pollock Pollachius (Figure 1; Research station in Venray water reduces the growth the infections. the Vibrio load in shrimp applications to P. acidilactici mayinclusion rate
in by 15 percent
enteric control of Leon Claessens plemented of the feed at an be able to weight have FCR and survival were evaluated
We gain, recently also begun to assess
for much of the past decade. pollachius (Gatesoupe 2002), channel catfish divided over 20 netssuch intestinal improve- leaching in fresh water This is particularly relevantwell-defined, prevent percent. Six pathogen for treatment the efficacy of this probiotic in group. The
and Not only could (volume 150L) in and reduces growth
aquaculture. AquaStar® Aquaculture Farming Technology0.5 or reduce replicates damage and
is a to salmo- of throughout the study for each tilapia at
average 1 9,91 11,36 Ictalurus punctatus
12,39 1,24 two separate tanks. Shrimp were acclima- by 33 percent. Email: info@aquaculture-ft.com used. During the trial gut of have been
nid culture as probiotic product tract fish translocation in the anterior all datasalmonids
multi-strain the gastrointestinal for is were Plymouth (Ferguson et70 days.
experiment lasted for al. 2010).
tised for several weeks to a lower salinity. Website: www.aquaculture-ft.com
However, we must not continue to use the (Shelby et al. 2007), potential port entry for beneficial and we are presently using novel approaches
a and shrimp andofpromotes a many fish recorded. We fed triplicate tanksthat AquaStar® was
The results showed of genetically male
average 2 9,70 11,13 12,25 1,28
umbrella term too loosely and be mindful that tilapia Oreochromis Each tank is connected to a biofloc Week one to six pathogens and both Vibrio as an improved at The University of Plymouth to test this tilapia (12 in 175g fish per tank) a basal diet
– overall
gut microflora as well anguillarum and Enterococcus faecium – as part of effective x controlling Vibrio parahaemolyti-
average 3 9,49 10,94 niloticus (Shelby et
not all probiotics display the same properties 12,30 1,41 reactor to control water quality. This way, performance (see Aeromonas salmonicida (the causative agents hypothesis. Hatchery - was found along the with or awithout the addition of and improved
table 5).
environmental condition in shrimp and AquaStar® cus in biofloc culture system P. acidilactici.
or yield the same subsequent benefits. al. 2006; Ferguson et the water quality is equal for all nets. One • Growth is negatively of vibriosis lowerfurunculosis, respectively)
affected by and
fish ponds. Indeed, a study supported by the
shrimp digestive system in the group fed the After 14 days feeding on the experimen-
the overall productivity of the system. Survival
EZ Artemia IAF2:Layout 1 6/16/2010 3:53 PM Page 1
Probiotics are inclusive of 10,42 range
average 4 a wide 11,62 al. 2010) and shrimp
12,68 1,13 tank was running on a salinity of 10ppt and salinity.
of microbes: Gram-positive bacteria, Gram- (Castex et al. 2008, the other on 5ppt. Each diet was used in • The best
negative bacteria, yeasts and cyanobacteria 2010) but the bene- three replicates for the 10ppt salinity and growth was
two replicates for the 5ppt salinity. observed Is artemia throwing you for a loop? Are you sure
have all shown some varying degree of fits to rainbow trout
Oncorhynchus Each net is equipped with a belt feeder. with diet The event that I‘m not missing
will energize
success when applied to fish. They provide
benefits to the host fish, in part at least, mykiss health and Feeding was adjusted daily to an estimated number a key essential
by improving the microbial balance and production is most growth and percentage of biomass. two for 10 nutrient?
functionality of the host gastrointestinal well documented and lower your BUSINESS
microbial communities. (Quentel et al. 2004; Results salinities.
Aubin et al. 2005a,b; During the first two weeks (see table 2)
Improving fish health Merrifield et al. the shrimp were fed continuously through Conclusion
From current literature, it is clear that 2010a,b). a belt feeder. Growth was good for all Growth VI INTERNATIONAL
replicates. Average growth loss between and survival AQUACULTURE
probiotic applications can improve fish We have previ-
health, disease resistance, growth perform- ously shown at 10ppt and 5ppt was 4.3 percent. are negatively New S yn th e tic Ar t e mia EXHIBITION
ance, feed utilisation, carcass composition, Plymouth that the The most promising diets were diet affected by for Shrimp from Z1 to PL
intestinal morphology and reduce mal- provision of dietary number two and four. For diet four lower salinity New
formations; however, we are yet to find a P. acidilactici can lead there was no growth diffference between of pond water
Formula
&
Smaller
VANNAGEN ®
specific probiotic candidate that can achieve to potentially resi- salinities. and by decreas- Size
all of these effects. dent populations These results show that Litopenaeus ing feeding � Up to 100% artemia replacement in shrimp trials naturally supports…
Some candidates are able to mediate in close association vannamei can easily be grown at good frequency. � No hatching or processing
growth rates in lower salinities if minerals The addi-
… Per formance
more benefits than others, and perhaps with the intestinal � Shortens larval culture cycle up to 3 days
are added in the diet. tion of various … Health
one of the most well studied, which we epithelium of rain- � Contains VpakTM and probiotics for stronger PL’s
have been working on at the Fish Nutrition bow trout, even For week three and four the feeding minerals in … Stress management October 20th to 23rd
� Completely Bi-secure
frequency was lowered to only four times feeds makes T Puerto Montt - Chile
and Health Research Aquarium, at The though other com- 50-2wo Size
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University of Plymouth, is Pediococcus aci- mercially available per day. This will result in slower feed it possible to -500 ron
www.chemoforma.com
uptake, longer interaction of feed and water still obtain Mic s
dilactici (Bactocell®, Lallemand). probiotics could ron
s
P. acidilactici strains produce a range of not (Merrifield and higher leaching of minerals from the growth in those
antimicrobial proteins (pediocins) and organic et al. 2010a,b). feed. The salinity of tank number two was conditions. Last available stands: book now!
María Paz / mpfernandez@aqua.cl / +56-2 756 5402 / +56-9 9547 7589
acids (such as lactic and acetic acid) which Subsequently, the Figure 1. TEM lowered from five to four ppt.
micrographs reveal that although Diet number
gives them antagonistic properties against a application of P. microvilli appear healthywasboth groups they are the
Growth in affected negatively by two seems to Zeigler Bros., Inc.
significantly longer feeding frequency and dropped to P. O. Box 95, Gardners, PA 17324 We have your per for mance in mind
range of Gram-positive and Gram-negative acidilactici enhanced lower in trout fed P. acidilactici (B) be the best 717-677-6181 SponSorS organizerS
than the control (A). Scale bar = 1 µm. (Merrifield et www.zeiglerfeed.com
bacteria, notably pathogenic Vibrio spp.. the gut ultrastruc- 1.25g/week (see table 3). source of min-
al. 2010b) Product of USA Chemoforma Ltd. CH-4302 Augst Switzerland
A clear effort to assess the use of this ture and improved There was no negative influence of erals in both Tel +41 61 811 33 55 Fax +41 61 811 28 03
nal AquAFeed | July-august 2010 18 | InternatIonal AquAFeed | July-august 2010
36 | InternatIonal AquAFeed | July-august 2010 July-august 2010 | InternatIonal AquAFeed | 15 July-august 2010 | InternatIonal AquAFeed | 19
July-august 2010 | InternatIonal AquAFeed | 37

3. F: Probiotic Fatten up your bottom line. Buhler high-performance animal and aqua feed production
systems are used by leading companies around the world. These producers know they
tal diets serum lysozyme activity and cir- sal interface which help
can rely not just on the technology itself, but also on the support that accompanies it. A
culating leucocyte levels were significantly mediate the reported
higher in the P. acidilactici group than the host benefits in order to
control group (Figure 2). maximise the benefits of
service combining local presence with global expertise both lowers feed mill operating
At day 32, PCR-DGGE demonstrated future applications.
clear modulations of gastrointestinal micro- costs and increases capacity utilization. To find out more, visit www.buhlergroup.com.
VIVChina 2010
bial profiles and the presence of P. acidilactici References
(OTU – Pa) in the probiotic fed fish (Figure
Aubin, J., Gatesoupe,
3). The level of lactic acid bacteria in F.-J., Labbe, L., Lebrun, L.
the intestine significantly increased from (2005a). Trial of probiotics
<0.001 percent in the control group to to prevent the vertebral
9.81 percent in the probiotic group. These column compression
lactic acid bacterial isolates in the probiotic syndrome in rainbow trout
group were confirmed as P. acidilactici by 16S (Oncorhynchus mykiss
rRNA sequencing. Walbaum). Aquacult. Res. 36,
758–767. September 6 - 8, 2010
Bühler AG, Feed & Biomass, CH-9240 Uzwil, Switzerland, T +41 71 955 11 11, F +41 71 955 28 96
Beijing
Aubin, J., Gatesoupe, F.J., fu.buz@buhlergroup.com, www.buhlergroup.com
Quentel, C., Forraz, M.,
"We are continuing to Facqueur, J.M., Rochet, B.,
Usache, V. (2005b). Étude
pursue our scientific de l'amélioration de la
rentabilité et de la sécurité
investigations to further
a 2010
sanitaire des filières truite
arc-en-ciel et bar par
IV Chin
our knowledge of the l'utilisation de probiotiques
d visit V
par voie alimentaire. Rapport
complex probiotic- Final, Convention N°008-
me an
03C, Contract OFIMER
host interactions at the
Co
N°A01959. 67 pp.
mucosal interface" Castex, M., Chim, L., Pham, Figure 3. 40-60% DGGE of the V3 region of the
D., Lemaire, P., Wabete, N., microbial profiles of the intestine of tilapia after
Nicolas, J.-L., Schmidely, P. feeding on probiotic P. acidilactici (Ferguson et al.
& Mariojouls, C. (2008) 2010)
Probiotic P. acidilactici
After reverting the probiotic group application in shrimp
back to the control diet P. acidilactici Litopenaeus stylirostris mykiss Walbaum). Aqua. Nutr. in press. DOI:
persisted within the gastrointestinal tract culture subject to vibriosis in New Caledonia. 10.1111/j.1365-2095.2009.00712.x.
www.viv.net
for up to 17 days. Light microscopy Aquaculture, 275, 182–193.
Merrifield, D. L., Harper, G., Baker, R. T. M., Ringø, E.,
revealed that the histological pattern of Castex, M., Lemaire, P., Wabete, N., Chim, L. Davies, S. J. (2010b). Possible influence of probiotic
the intestinal tract did not differ between (2010) Effect of probiotic Pediococcus acidilactici adhesion to intestinal mucosa on the activity and
the control and probiotic groups: no signs on antioxidant defences and oxidative stress morphology of rainbow trout (Oncorhynchus
of tissue or cell damage were observed status of Litopenaeus stylirostris. Under Vibrio mykiss) enterocytes. Aquacult. Res. in press DOI:
and leucocyte infiltration levels remained nigripulchritudo challenge. Fish and Shellfish 10.1111/j.1365-2109.2009.02397.x.
Immunology 28, 622-631.
The international platform
unaffected. The simultaneous ability to Shelby, R.A., Lim, C., Yildirim-Aksoy, M. & Delaney,
elevate innate immune responses and Ferguson R. M. W., Merrifield, D. L., Harper, M.A. (2006) Effects of probiotic supplements on
fortify intestinal microbial communities G. M., Rawling, M. D., Mustafa, S., Picchietti, S., disease resistance and immune response of young
is likely to boost tilapia defences against
enteric infection.
Balcázar, J. L. &. Davies S. J. (2010) The effect of
Pediococcus acidilactici on the gut microbiota
Nile tilapia, Oreochromis niloticus. J. Appl. Aqua.,
18, 22–34.
for the Chinese Feed to Meat industry
From the present data it is clear that and immune status of on-growing red tilapia
Shelby, R.A., Lim, C., Yildirim-Aksoy, M. & Klesius,
there is a great potential for P. acidilac- (Oreochromis niloticus). J. Appl. Microbiol., early
P.H. (2007) Effects of probiotic bacteria as dietary
tici applications for intensively reared fresh view doi:10.1111/j.1365-2672.2010.04713.x
supplements on growth and disease resistance
water fish species of economic importance, Gatesoupe, F.-J. (2002) Probiotic and in young Channel catfish, Ictalurus punctatus
and the current applications conducted at formaldehyde treatments of artemia nauplii (Rafinesque). J. Appl. Aqua., 19, 81–91.
New CIEC, Beijing
the farm level will highlight the financial as food for larval pollack, Pollachius pollachius.
Quentel, C., Gatesoupe, F.J., Lamour, F., Abiven,
benefits of probiotic P. acidilactici use in Aquaculture, 212, 347–360.
A., Baud, M., Aubin, J. (2004) Effects of oral
industrial fish farming. Merrifield, D.L., Bradley, G., Harper, G.M., administration of probiotics on the resistance
At the University of Plymouth, under Baker, R.T.M., Munn, C.B., Davies, S. J. (2010a). of rainbow trout, Oncorhynchus mykiss, against
Professor Simon Davies, we are continuing Assessment of the effects of vegetative and Yersinia ruckeri: asymptomatic carriers and
to pursue our scientific investigations to lyophilised Pediococcus acidilactici on growth, humoral immune parameters. 6th Symposium on
further our knowledge of the complex feed utilisation, intestinal colonisation and health Fish Immunology, 26–29 May 2004. Åbo/Turku,
probiotic-host interactions at the muco- parameters of rainbow trout (Oncorhynchus Finland, p. 60. book of abstracts.
The solution behind the solution.
38 | InternatIonal AquAFeed | July-august 2010

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