BACILLUS AS AQUACULTURE PROBIOTICS: POTENTIAL AND … · Advantages of Bacillus Bacillus spores can...
Transcript of BACILLUS AS AQUACULTURE PROBIOTICS: POTENTIAL AND … · Advantages of Bacillus Bacillus spores can...
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BACILLUS AS AQUACULTURE PROBIOTICS: POTENTIAL AND LIMITATIONS
O. DecampINVE TECHNOLOGIES nv, Belgium.
D.J.W. MoriartyThe University of Queensland, AustraliaINVE Aquaculture Health, Thailand.
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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
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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
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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
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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
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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
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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
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Thank you