July | August 2010

Feature title: The Probiotic approach to elevating health of fish

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 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 (Officethis
diet including National
probiotic Interprofessionnel
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
number of de
Vibrio la Mer
spp. et
found de inl'Aquaculture),
the hepatopan-
salinity feed day 14 day 21 day 28 (g/week)
tion) could be the result of a lower solubil- growth was obtained with diet number Based on these results a special intestine
mineral of shrimp demonstrated
creas and that
intestine of P. acidilactici
shrimp fed AquaStarcould®
Control AquaStar®

The Probiotic approach

ity of the mineral; which could affect the three. premix was developed jointly by companies A study by Dr Kidchakan improve Hatcheryrainbow
was lower troutthanresistance
control group. to yers-
digestibility of this minerals for the shrimp. For week five and six the feeding fre- Prayon and CreveTec. Supamattaya (2006) at the iniosis, In aasrecent
demonstrated
study, thebyMarine reduced accu-of
Station
10 ppt 1 10,05 11,68 12,82 1,39 Therefore, the apparent digestibility was quency was further lowered to only two
Survival (%) 52.0a 83.0b Prince of Songkla University, mulated
Aquaculturemortalities
at theafter an
Federal intraperitoneal
University of
tested for some selected growth trials. It times per day. Thailand, has shown that the use injection
Rio challenge.
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 theAseffect
well asof benefits to intestinal
the simultaneous morphol-of
application
Final weight (g) 8.42a 9.05a
digestibility of selected minerals is not the on growth, the feed for the three replicates is effective in reducing the total ogy ® Pondresistance,
and disease
AquaStar and AquaStar ® Growout
P. acidilactici canin
10 ppt 3 9,79 11,24 12,44 1,33
problem. in tank number one was Final treatedbiomass (kg/tank)
before 45.97a
more inFormation:
78.87b
number of Vibrio bacteria found also alleviate
Litopenaeus rainbow
vannamei trout
cultured vertebral
in a col-
biofloc
Figure 2. Probiotic P.
Molecular
10 ppt profiling
4 of gastro-intestinal
10,83 11,74 13,02biota and 1,1 effects on gut ultra-structural characteristicsfeeding: Four nets received normal feeds;
FCR 2.70a
Frederic Martens
1.40b in the hepatopancreas and intes- umn
acidilactici significantly
compression
technology system syndrome
contaminated (VCCS) and
with Vibrio
Material and methods four nets received feeds whichProductivity were first(kg/m²)Prayon SA, 1.31a 2.25b
Rue J Wauters 144 tine of white
elevated the circulating shrimp (Panaeus improve aesthetic
parahaemolyticus quality
(Krummenauer by lowering
et al., con-
2009).
4 ppt 1 9,78 11,05 11,97 1,1 From the leaching experiment, the most soaked in fresh water for one a, bhour
Means within a row4480
before withEngis, Belgium
different vannamei),
superscripts differleucocyte levels which
andcan serumreduce the dition Thefactor (Aubin
juvenile whiteet shrimp
al. 2005a; wereMerrifield
stocked
by Dr Daniel L Merrifield and Matthew Emery, Senior microbiology technician,promising The School of Biomedical and Biologicalfeeding and four nets received feeds which Email: fmartens@prayon.be
combinations were used for a significantly (p < 0.05); analyzed by one-way analysis oflysozyme risk ofactivity
infectionof(see tilapia
Table 1). etinal. 2010a).
tanks at theThese
high benefits
density ofwhich improve
300 shrimp/m².
Sciences,
4 pptThe University2 of8,64
Plymouth, Plymouth,
10,06 11,38UK, Email: 1,37Daniel.merrifield@plymouth.ac.uk Tel: +32 4 2739310 after 14 days
feed trial. Four diets were produced on were first soaked in tank water (10ppt) for variance (ANOVA).
Website: www.prayon.com Groups of 20feeding
juvenile white the quality Feed (38 of the final product
percent could effec-
crude protein) was
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
suppliedboostthreeproduction
times/day. andThe elevate trout
experimental
millimeters. There was a clear influenceeffectiveofatleaching
inhibiting a wideEric Derange
Muylderof fish into 200L glass aquaria and fed farming profit margins.
group additionally received three grams of
4 ppt 4 10,00 11,50 12,34 1,17 Shrimp (Litopenaeus vannamei) were on growth, indicating that leaching of nutri- CreveTec

P
pathogens.
ments provide a foundation for enhanced to
can satiation
infect and five times
cause daily
severe during
tissue a six-week
damage to AquaStar ® Growout/kg feed and 0.5ppm/
Email: eric@crevetec.be
10 ppt
robiotics and their applica-
average 10,36 11,71
probiotic
12,85
in fish culture
1,25
has been under-imported from
microvilli Thailand
morphology and raised
of the to two
enterocytesents is an imported factor in the
Recent
digestive growth
data
function from
but in vivo
maintaining experiments
epithelial
Website: www.crevetec.be
period.
the A
anterior commercial
intestine of type
salmonid diet was
fish. used
From The financial benefits
week of AquaStar ® Pond during the rear-
tions in fish culture have taken; information regarding the benefits ofgrams.inThey were then
the anterior transferred
intestine to the troutof shrimp. Leaching for integrity
of rainbow one houris
suggest inparamount
that 10ppt
AquaStar®tomay defend against
be beneficial as a control.
these findings AquaStar ® Hatchery
we hypothesise that was sup- of
dietary ing this probiotic
period. Biological parameters, growth,
4 ppt been studied
average and discussed
9,40 10,81 P. acidilactici
11,96 is available
1,28 for pollock CreveTec-AFT
Pollachius (Figure 1; Research
Merrifield station
et al. in
2010b).Venray water reduces the growth
enteric
in by
the 15 percent
infections.
control of the Vibrio load in
Leon Claessens shrimp applications
plemented of
to P.
theacidilactici
feed at may
an be able
inclusion to
rate weight gain, FCR and survival
We have recently also begun to assess were evaluated
for much of the past decade. pollachius (Gatesoupe 2002), channel catfishand divided Notoveronly 20 netssuch
could (volume 150L)improve-
intestinal in and leaching in fresh water reduces
This growth
is particularly
aquaculture. ® Aquaculture
AquaStarrelevant to Farming prevent
salmo- Technology
is a well-defined, or reduce
of 0.5 percent. Six pathogen
replicates for damage and
treatment throughout
the efficacy of thethis
study for each
probiotic in group.
tilapia Theat
average 1 9,91 11,36 Ictalurus
12,39 punctatus1,24 two separate tanks. Shrimp were acclima- by 33 percent. nid culture as probiotic
multi-strain Email: info@aquaculture-ft.com
the gastrointestinal
product tract
for fishis translocation in the anterior
were used. During the trial allgutdata
of salmonids
have been Plymouthexperiment lasted foret70
(Ferguson al.days.
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), a and
potential
shrimpport andofpromotes
entry for a many fish
beneficial and we are presently using novel approaches
recorded. WeThefedresults showed
triplicate tanksthat AquaStar®male
of genetically was
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 –gutoverall and both
microflora as Vibrio
well as anguillarum
an improved and at The University offaecium
Enterococcus Plymouth – to as test
partthisof tilapia (12 inx 175g
effective fish per
controlling Vibriotank)parahaemolyti-
a basal diet
not all probiotics display3 the same
average 9,49 10,94 niloticus
properties 12,30 (Shelby et1,41 reactor to control water quality. This way, performance (see Aeromonas table 5). salmonicida
environmental (the causative
condition in shrimp AquaStar® Hatchery - was found along the with
and hypothesis.
agents cus orin awithout
biofloc the addition
culture system of and improved
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 affected by lower
fishvibriosis
ponds. and furunculosis, respectively) Indeed,
shrimp a study
digestive systemsupported
in the groupbyfedthe the theAfter 14 productivity
overall days feedingofon thethe experimen-
system. Survival
average are inclusive4 EZ Artemia IAF2:Layout 1 6/16/2010 3:53 PM Page 1
Probiotics of 10,42
a wide range11,62 al. 2010)
12,68and shrimp1,13 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
300 00 Mic s!
www.aqua-sur.cl
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
micrographsfrom five to four
reveal thatppt.although Diet number
gives them antagonistic properties against a application of P. microvilli appearGrowth
healthywas in both groups
affected they are
negatively by the two seems to Zeigler Bros., Inc.
significantly longer in trout frequency
fed P. acidilactici (B) 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 feeding and dropped 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 | InternatIonalAquAFeed

18| InternatIonal
36 AquAFeed| July-august
| July-august2010
2010 July-august 2010 | InternatIonal AquAFeed | 15 July-august2010
July-august 2010| InternatIonal AquAFeed| 37
| InternatIonalAquAFeed | 19
 F: Probiotic
tal diets serum lysozyme activity and cir-
culating leucocyte levels were significantly
higher in the P. acidilactici group than the
control group (Figure 2).
At day 32, PCR-DGGE demonstrated
clear modulations of gastrointestinal micro-
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
sal interface which help
can rely not just on the technology itself, but also on the support that accompanies it. A
mediate the reported
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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
1 0
sanitaire des filières truite

our knowledge of the

arc-en-ciel et bar par
l'utilisation de probiotiques

C h i n a 2 0
I V
d visit V
par voie alimentaire. Rapport
complex probiotic- Final, Convention N°008-

host interactions at the

mucosal interface"
03C, Contract OFIMER
N°A01959. 67 pp.

After reverting the probiotic group
back to the control diet P. acidilactici
persisted within the gastrointestinal tract
Castex, M., Chim, L., Pham,
D., Lemaire, P., Wabete, N.,
Nicolas, J.-L., Schmidely, P.
& Mariojouls, C. (2008)
Probiotic P. acidilactici
application in shrimp
Litopenaeus stylirostris
culture subject to vibriosis in New Caledonia.
Figure 3. 40-60% DGGE of the V3 region of the Co m e a n
microbial profiles of the intestine of tilapia after
feeding on probiotic P. acidilactici (Ferguson et al.
2010)
mykiss Walbaum). Aqua. Nutr. in press. DOI:
10.1111/j.1365-2095.2009.00712.x.

for up to 17 days. Light microscopy
revealed that the histological pattern of
the intestinal tract did not differ between
the control and probiotic groups: no signs
of tissue or cell damage were observed
and leucocyte infiltration levels remained
unaffected. The simultaneous ability to
elevate innate immune responses and
fortify intestinal microbial communities
is likely to boost tilapia defences against
enteric infection.
From the present data it is clear that
there is a great potential for P. acidilac-
tici applications for intensively reared fresh
water fish species of economic importance,
and the current applications conducted at
Aquaculture, 275, 182–193.
Castex, M., Lemaire, P., Wabete, N., Chim, L.
(2010) Effect of probiotic Pediococcus acidilactici
on antioxidant defences and oxidative stress
status of Litopenaeus stylirostris. Under Vibrio
nigripulchritudo challenge. Fish and Shellfish
Immunology 28, 622-631.
Ferguson R. M. W., Merrifield, D. L., Harper,
G. M., Rawling, M. D., Mustafa, S., Picchietti, S.,
Balcázar, J. L. &. Davies S. J. (2010) The effect of
Pediococcus acidilactici on the gut microbiota
and immune status of on-growing red tilapia
(Oreochromis niloticus). J. Appl. Microbiol., early
view doi:10.1111/j.1365-2672.2010.04713.x
Gatesoupe, F.-J. (2002) Probiotic and
formaldehyde treatments of artemia nauplii
www.viv.net
Merrifield, D. L., Harper, G., Baker, R. T. M., Ringø, E.,
Davies, S. J. (2010b). Possible influence of probiotic
adhesion to intestinal mucosa on the activity and
morphology of rainbow trout (Oncorhynchus
mykiss) enterocytes. Aquacult. Res. in press DOI:
10.1111/j.1365-2109.2009.02397.x.
The international platform
Shelby, R.A., Lim, C., Yildirim-Aksoy, M. & Delaney,
M.A. (2006) Effects of probiotic supplements on
disease resistance and immune response of young
Nile tilapia, Oreochromis niloticus. J. Appl. Aqua.,
18, 22–34.
for the Chinese Feed to Meat industry
Shelby, R.A., Lim, C., Yildirim-Aksoy, M. & Klesius,
P.H. (2007) Effects of probiotic bacteria as dietary
supplements on growth and disease resistance
in young Channel catfish, Ictalurus punctatus
(Rafinesque). J. Appl. Aqua., 19, 81–91.

New CIEC, Beijing

the farm level will highlight the financial
benefits of probiotic P. acidilactici use in
industrial fish farming.
At the University of Plymouth, under
Professor Simon Davies, we are continuing
to pursue our scientific investigations to
further our knowledge of the complex
probiotic-host interactions at the muco-
38 | InternatIonal AquAFeed | July-august 2010
as food for larval pollack, Pollachius pollachius.
Aquaculture, 212, 347–360.
Merrifield, D.L., Bradley, G., Harper, G.M.,
Baker, R.T.M., Munn, C.B., Davies, S. J. (2010a).
Assessment of the effects of vegetative and
lyophilised Pediococcus acidilactici on growth,
feed utilisation, intestinal colonisation and health
parameters of rainbow trout (Oncorhynchus
Quentel, C., Gatesoupe, F.J., Lamour, F., Abiven,
A., Baud, M., Aubin, J. (2004) Effects of oral
administration of probiotics on the resistance
of rainbow trout, Oncorhynchus mykiss, against
Yersinia ruckeri: asymptomatic carriers and
humoral immune parameters. 6th Symposium on
Fish Immunology, 26–29 May 2004. Åbo/Turku,
Finland, p. 60. book of abstracts.
The solution behind the solution.
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