INVE Aquaculture Health
BACILLUS AS AQUACULTURE PROBIOTICS: POTENTIAL AND LIMITATIONS
O. DecampINVE TECHNOLOGIES nv, Belgium.
D.J.W. MoriartyThe University of Queensland, AustraliaINVE Aquaculture Health, Thailand.
INVE Aquaculture Health
Bacillus as probiotic for animal use
Authorised additive EUBacillus cereus var. toyoi (Toyocerin)Bacillus subtilis (Belfeed, Calsporin, Bioplus 2B)Bacillus licheniformis (Bioplus 2B)http://www.efsa.eu.int/science/feedap/an_applications/catindex_fr.html
No authorised additive EUMany products authorised Asia/Latin AmericaBacillus cereus var. toyoiBacillus coagulansBacillus lentusBacillus subtilisBacillus pumilusBacillus licheniformis…
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Benefits of selected Bacillus strains
• Present in aquatic animals and environments• Ability to grow under wide range of conditions• Producer of numerous enzymes• Active against many pathogens
• Directly by growth inhibition• Indirectly by competitive exclusion
• Advantages of spores• Long shelf life • Easy to transport• Resistant to dehydration • Resistant to temperatures up to 60° C
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PROBIOTICS AS ALTERNATIVE TO PROPHYLACTIC USEOF ANTIMICROBIALS: LIMITATIONS AND POTENTIAL
•Doubts of efficacy
•Doubts of composition
•Doubts of safety
•Suitable claims
•Suitable delivery
•Suitable QC
•Suitable production
•Suitable registration
INVE Aquaculture Health
Advantages of Bacillus mixtures
Doubts of compositionsafety
Germinationnot fermentation
<8 hours for activationgermination
24 hours for fermentation
Bacillus Spores high concentration
Freeze-driedor liquid culture low concentration
contamination
INVE Aquaculture Health
PROBIOTICS AS ALTERNATIVE TO PROPHYLACTIC USEOF ANTIMICROBIALS: LIMITATIONS AND POTENTIAL
Registration
Safety to environment
Safety to target
Formulations – COST/PRACTICALITIES
Screening of strains for function
R&D Farmers
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Function: from lab to pilot-scale
Field testSanolife MIC
Lab “Pathogen control”
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SAFETY: environment including humans
• Non-target aquaculture organisms
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SAFETY: Survival of starved seabream larvae
Surv
ival
of
seab
ream
larv
ae (%
)
Days after hatching
0
20
40
60
80
100
0 2 4 6 8 10
2A
3
LMG
i8
10i
isol
anm
INVE Aquaculture Health
SAFETY: Survival of starved seabream larvae
0
20
40
60
80
100
0 2 4 6 8 10 12 14DAH
Surv
ival
(%)
Bacillus mixture conc1
Bacillus mixture conc2
Filtered w ater
Sterile w ater
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SAFETY: environment including humans
• Humans
Bacillus cereus
B. cereus enterotoxins
emetic toxin genes
INVE Aquaculture Health
PROBIOTICS AS ALTERNATIVE TO PROPHYLACTIC USEOF ANTIMICROBIALS: LIMITATIONS AND POTENTIAL
Sales
Registration
QC
Upscale production – COST?
Safety issues
Commercial
R&D
Support
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UPSCALE PRODUCTION
Advantage of Bacillus
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?Species with little history
UPSCALE PRODUCTION
Cost!!!
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Feed back from field
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Shrimp Grow-Out Probiotics
Sanolife PRO-1/PRO-2 mixed with feed (min 108 cfu/g feed)
$0
$400
$800
$1.200
$1.600
$2.000
$2.400
$2.800
Controls5-7
Treated2,3 & 4
Treated2 & 3
US
Dol
lars
0
20000
40000
60000
80000
100000
120000
140000
Indi
an R
upee
s
INVE Aquaculture Health
Alternative delivery – incorporation in feed
Advantages of Bacillus
Bacillus spores can exist in a desiccated and dehydrated state almost indefinitely.
The resistance of spores can be explained by various factors:- thick proteinaceous spore coats- reduced permeability of the spore core- reduced water content - etc (Setlow 1999)
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Alternative delivery – incorporation in feed
“The extrusion-expansion and drying process resulted in the loss of >99 % of spores” Biourge et al. 1998. The use of probiotics in the diet of dogs. J Nutr. 1998 Dec;128(12 Suppl):2730S-2732S.
Ruiz et al. J. Food Sci. 67(2)
Selection of resistant strains
What about claims?
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Alternative delivery – incorporation in feed
0,0
0,2
0,4
0,6
0,8
1,0
1,2
1,4
1,6
1,8
-5 0 5 10 15
Storage time (month)
Spor
e co
ncen
tratio
n (1
0^6
cfu/
g)
Batch 1
Batch 2
Batch 3
Batch 4
Biourge et al. 1998. J. Nutrition 128
1. Loss during production
-20°C 4°C Room 30°C
1,E+03
1,E+04
1,E+05
1,E+06
1,E+07
1,E+08
1 month storage Sanolife PRO-1 temperature
2. Loss during storage
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FURTHER INVESTIGATION
-Legal status
-Spore germination, proliferation, colonisation
- Resistant pathogens (geographical variations, history, management, etc)
- Follow-up of probiotics
INVE Aquaculture Health
Thank you
