The dietaryThe dietaryPROBIOTIC to PROBIOTIC to
secure the health secure the health of of shrimpsshrimps
• In hatcheries, Bactocell is particularly indicated to:– Maintain a healthy microbial environment – Improve larvae quality (reduce deformities) and
development (facilitate transition from Zoe to Mysis)– Improve resistance of larvae to opportunistic pathogens &
stressSuppress use of antimicrobial compounds
� A solution for high larval production
BACTOCELL applications in BACTOCELL applications in HatcheriesHatcheries
BACTOCELL® 100
The The productproduct
Concentrated formula PA100 for hatcheries(100 billion CFU/g (10x1010 CFU/g))
-Standard dosage:- Larval tanks: 1g/m3 of water- Live preys: 1 to 5g/m3 of culture water
The strain: Pediococcus acidilactici MA18/5M
Yeast MouldBacilles GRAM+
sporulated
Bacilles GRAM+
no-sporulated
Bacillus LactobacillaceaeEnterococcus
FungiBacteria
Pediococcus
MICROORGANISM
Coques GRAM+
no-sporulated
Pediococcus and other probiotics microorganisms classification
Pediococcus acidilacticiPediococcus acidilacticiMA 18/5MMA 18/5M
The The strainstrain
The The strainstrain
BACTOCELL ADMINISTRATION
Live Prey
Microparticules (transition feeds)
Nursery feeds
Larval feeds
Enrichments
Grow out conditions of use:
100-200g/kg PA10
1-5g.m3of the PA
100
Water
1g.m3 of the PA
100
5-10g.m3 Direct mixing: 10-20g/kg of
PA 100
The The productproduct
Larval tank Nursery tanks
TA
NK
SF
EE
DIN
G
Preparation dose Bactocell 100 at 0.1g.m-3
Preventive dose Bactocell 100 added directly in tank at 1g.m-3 every day
Tank filling PL5Zoe Mysis
Microencapsulated diet
Bactocell PA10 mix with feed: 1g.kg-1
Complete feedBactocell PA10 mix with feed: 2g.kg-1
A0
Artemia hatching tanks: Bactocell 100 at 5g.m-3
Feeding programs for shrimps - hatcheryBACTOCELL applications in AquacultureBACTOCELL applications in Aquaculture
Bactocell is active in seawater
1,0E+02
1,0E+03
1,0E+04
1,0E+05
1,0E+06
S11 S17 S22 S27
Total Marine Microflora (CFU/g)
Vibrio Flora (CFU/g)
Bactocell PA100 (CFU/g)
In sea water tanks Bactocell is active :- It ranged 10e4-10e5 CFU/ml seawater- Vibrio level are below the detection threshold
* : belowdetectionthreshold2.10e3 CFU/g
* * * *
Population in Water tanks (CFU /ml)
The The productproduct
Bactocell is active in Enriched Live preys
• Bactocell concentration in artemia enriched during 14 hours (Gatesoupe, 2002)
• Bactocell concentration in rotifers:– Use in the culture� (Marine
hatchery)– Use during enrichment only
(2 hours)� Bactocell reach 10e4-10e5
CFU/ml (IFREMER, 2010)
The The productproduct
Bactocell reaches concentration between 10e4-10e5 CF U/ml
6.8 E+037.6 E+06Day 2
1.4 E+042.1 E+03Day 1
2.5 E+051.2 E+04Day 0
Rotifers (CFU/ml)Culture water (CFU/ml)
Day of culture
6.8 E+037.6 E+06Day 2
1.4 E+042.1 E+03Day 1
2.5 E+051.2 E+04Day 0
Rotifers (CFU/ml)Culture water (CFU/ml)
Day of culture
Bactocell administration to larvae through level preys is efficient
The The productproduct
Bactocell reaches concentration between 10e4-10e5 CFU/g of gut
1,0E+02
1,0E+03
1,0E+04
1,0E+05
1,0E+06
1,0E+07
S11 S17 Gut
Total Marine Microflora (CFU/g)
Vibrio Flora (CFU/g)
Bactocell PA100 (CFU/g)
Microbial population in the gut
Bactocell count in the gut
Direct administration to larvae through feed or water is also anefficient way to deliver Bactocell in hatcheries
The The productproduct
Bactocell reaches again concentration between 10e4-1 0e5 CFU/g of gut
Stomach Digestive gland Gut
Bac
teria
l con
cent
ratio
n (C
FU
.g-1
)
1e+2
1e+3
1e+4
1e+5
1e+6
1e+7
1e+8
1e+9
1e+10
1e+11P. acidilacticiFlore totaleVibrio
Stomach Digestive gland Gut
P. a
cidi
lact
ici
(CF
U.o
rgan
-1)
1e+2
1e+3
1e+4
1e+5
*
IFREMER, 2009
Administration through the feed
Direct administration to larvae through feed or water is also anefficient way to deliver Bactocell in hatcheries
The The productproduct
Bactocell reaches again concentration between 10e4-1 0e5 CFU/g of gut
Villamil et al., 2010
Administration through water
Protocol for Artemia Enrichment• Protocol defined based on the work conducted with the IFREMER
(Gatesoupe, 2002)
– Bactocell solution preparation • Weigh the right amount of Bactocell in a bucket• Add the sea water, natural salinity (no chlorinated water) • Stir for 2-3 minutes with the enrichment product • Check the consistency of the mixture. • Distribute directly the solution at the water inlet tofavor dilution in the tank
– Enrichment step:• Cysts are decapsulated and kept 24 to hatch, • we obtain AO (nauplii) which are then rinsed 20min. • Enrichment:
– 300 A0 / ml– Salinity natural – 14 to 24 hours enrichment – Recommended dose: the objective is to work 5.10exp 5 CFU / ml of enrichment medium for
300 A0 / ml – Dosage = 5g Bactocell per m3 of enrichment media (In the case of bad quality culture In the case of bad quality culture
BactocellBactocell can be increase up to 10g/mcan be increase up to 10g/m 33))• Then follow the standard enrichment final procedure and distribution protocol
The The productproduct
The The strainstrain
BACTOCELL EFFECTS
Pediococcus acidilactici MA18/5M prevents the colonization and development of pathogenic bacteria and
positively influence microbial communities
The The strainstrain
Prevention of pathogenic bacteria development (Vibrio, pseudomonas,…)
- Acidification of pathogens interior- Metabolism perturbations
Bactocell
Lactic acid
Pathogens
Pathogens growth
XpH
• Antagonism against Vibrio:– in vitro
– In vivo
Ex: Live feed afterenrichment with Bactocell:Decraese in Vibrio countaby 10 to 100 times
ExperimentalExperimental studiesstudies
Number of active live preys/ non mobile ones
Bactocell cycles� direct benefits
• Antagonism against Vibrio:– In vivoEx: Live feed culture with Bactocell: Decrease in Vibrio count
by 10 to 100 times
ExperimentalExperimental studiesstudies
Vib
rio (
logC
FU
.g-1
)
1e+7
1e+8
BactocellControl
Before feeding After feeding
Vib
rio (
logC
FU
.g-1
)
1e+7
1e+8
BactocellControl
Before feeding After feeding
P<0,05
ExperimentalExperimental studiesstudies
• Antagonism against Vibrio:– In vivo
Ex: Decrease in Vibrio concentration in the gut of the shrimps after feeding with Bactocell
Bactocell group
L.stylirostris, IFREMERP. Monodon
Aquac. Sc. Inst. CHINA
LOCATION : DAC-IFREMER New Caledonia/ 2005.
OBJECTIVE : Bactocell evaluation as antibiotic replacement in larval production
EXPERIMENTATION
Administration mode : directly into the waterInclusion rate : 3 doses: 107 CFU/100l (L1), 2.107/100l CFU/g (L2) and 3.107 CFU/ 100l (L3)Duration: 10 daysRearing system : 500L tanks and cages in pondDensity: Feeding frequency:
ANIMAL
Specie : Larvae of Litopenaeus stylirostrisStade of development: from mysis to nauplii
(Pham D., 2005)
ExperimentalExperimental studiesstudies
0%
20%
40%
60%
80%
100%
120%
E L1 L2 L3 T-Treatment
Sur
viva
l (%
)
J0 J4 J6 J9
• Effects on survival:– A significant dose effect of Bactocell– Bactocell at 3.107 CFU/100l lead to the same results
than antibiotic treatment (E: erythromycine, T- : control)
ExperimentalExperimental studiesstudies
• Effect on larvae developement:– Bactocell doses between 2.107 CFU/100l and 3.107
CFU/100l allows to enhance the proportion of mysisstage compared to the control.
Proportion of the various larval stage for the diff erent treatment after 9 days
0%
20%
40%
60%
80%
100%
120%
140%
160%
T- L1 L2 L3 E
Treatment
Pro
porti
on %
ZoéIII Mysis I Mysis II Mysis III
ExperimentalExperimental studiesstudies
LOCATION : Aquaculture Science Institute Shengzhen DapengTrial Center, China, 2003
OBJECTIVE: Bactocell evaluation on intensive breading type experiments
EXPERIMENTATION
Administration mode : via the feedInclusion rate : 106 CFU/g of feedDuration: 7 weeksRearing system : 2,5-m³ tanks Density: 2500 PL 15/m³
ANIMALS
Specie : Penaeus monodonStade of development : PL 15
ExperimentalExperimental studiesstudies
50
60
70
80
90
100
110
CONTROL BACTOCELL
% S
urvi
val
CONTROL
BACTOCELL
94,6
73,3
0,2
0,3
0,4
0,5
0,6
0,7
0,8
0,9
1
1,1
Indi
vidu
al W
eigh
t (g)
CONTROL
BACTOCELL
0,4
0,8
• Effects on survival– Increases survival by
29% (P<0,05) after 7 weeks.
• Effects on growthparameters– Significant increase in
weight by 50%(P<0,05)
– Non significant increases in length by 16% (P>0,05)
ExperimentalExperimental studiesstudies
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