Microbial management in fish and shrimp on growing

1. Microbial management in fish and shrimp on growing Rudi Bijnens Ridley Agriculture Prawn and Barramundi association conference

2. Challenges facing aquaculture producers • Pathogens have a higher change of creating disease • More feed and animals • Deteriorating water and pond bottom quality Environment Pathogen Host • Higher stocking densities • will increase stress

3. Meeting these challenges Environment Pathogen Host INVE Aquaculture health Management based on Prevention

4. Bio security - Definition “An effective biosecurityprogram requires an understanding of the aquaculture production systems, the principles of disease transmission, the biology of the reared animals, the advantages and limitations of the products used and management techniques adopted.” Moriarty & Decamp 2009. The Rising Tide pp 53-70 “…sets of practices that will reduce the probability of a pathogen introduction and its subsequent spread from one place to another…” . Lotz 1997. World J MicrobiolBiotechnol 13:405–413

5. Sanocare PUR The highly effective disinfectant and virucide for aquaculture. Multiple mode of action • High dosage (1%) for total kill • Cleaning of equipment • Foot baths • Lower dose (0.3 – 1 ppm) for bacteriostatic action • Control pathogens during culture • Cure skin infections of prawn/fish

6. Mode of action Cl- + O2 + OH- 2. Sulphamic acid Potassium monopersulfate Organic acids  low pH (1 %) + virucidal Surfactant  allow disinfectant to penetrate organic matter Sodium chloride + potassium monopersulfate + sulphamic acid  production of free chlorine 3. Hydrolysis Potassium monopersulfate Oxidizing agent

7. Biofilm of Vibrio – resistance to antimicrobials Failure of an agent to penetrate the full depth of the biofilm. Polymeric substances in the matrix of a biofilm retard the diffusion of chemicals. Costerton et al. 1999. Science 284:1318-1322 biocide Not affected by the biocide

8. High dosage application (1%) vsbiofilm

9. Low dosage application vs fungi In vitro effect on hyphal growth (Lagedinium sp.) • Fungistatic at low dose • Not a fungicide (like Treflan) Results

10. Biosecurity – destruction of viruses • 30ppm in 3 hours contact time sufficient to kill WSSV • Difficult to reproduce conditions from ponds with longer contact time • Confirmed efficacy in pond preparation

11. Sanocare PUR Recommended protocols of use 1% (0.8Kg in 80L) to disinfect pond bottom and side before stocking (prawn + Barramundi) 1% to disinfect area surrounding diseased pond to prevent disease spreading to the rest of the farm 0.5ppm to control bacteria during the crop 0.5ppm to clean prawn exoskeleton before harvest (against algal fouling and lesions)

12. Probiotic’s - Definition • Historically, probiotics were defined according to their expected benefit to the host’s intestinal balance. • In aquaculture, extension of the definition considering the level of interaction between the rearing medium and the animal. Traditional Definition + Bioremediation = Microbial technology. • Live microbial additive with a beneficial effect on the host by: • modifying the host-associated /ambient microbial community • ensuring improved use of the feed • enhancing the nutritional value of the feed • enhancing the host response towards disease • improving the quality of its ambient environment

13. Sanolife PRO-2, PRO-F and PRO-W Trueprobiotics for pond based farming shrimp and fish 5 x1010cfu/g • Applied to the water • Active in the water and the pond bottom • Recycling waste into nutrients and biomass • Resulting in stable phytoplankton bloom and reduced ammonia levels • (off flavour Tilapia) 2 x1010cfu/g Coated on the feed Active in the gut Inhibiting pathogens supporting food digestion resulting in better growth and lower FCR

14. Selection of bacillus strain • Safety • Natural for the animals and their environment • No pathogenic for animals or humans • Technological requirements • Cost-effective manufacturing • consistent product • long shelf life • Functional requirements • Able to perform in all aquaculture environments: waste degradation, support hosts’ digestion, inhibition of pathogens • Selection of specific Bacillus strains that are adapted to a wide range of aquaculture environments

15. Inhibition of pathogens • Numerous publications on in vitro inhibitions • Strains of Arthobacter, Bacillus, Lactobacillus, Pseudomonas, Saccharomyces, Streptococcus, Vibrio, etc. • Recent isolates or strains from other applications 1 2 3 Check inhibition

16. Functional requirements • Persistence in shrimp or fish intestinal tracts: • Displace and inhibit vibrios • Provokes reaction of immune system • Produce enzymes that support feed digestion: FCR • Importance of feeding frequency

17. Probiotics. Improved FCR HOW? Better use of carbohydrates explained by a stimulation of alpha amylase. Effect on metabolism Source of nutrients or growth factors Bacillus feed probiotics. Ecuador Bacillus feed probiotics. Brazil

18. Waste management Bacillus spores Phytoplankton Feeds Zooplankton Bacillus metabolites Host Bacillus exo-enzymes Bacillus Waste Pond bottom

19. Proof of concept Waste management • Water and pond bottom quality improves • Less stress for the animals  Survival & Growth • Reduced need for water change • zero water exchange? • Recycle pond water into next crop? • Waste is recycled into nutrients and single cell proteins which are turned into live food  FCR Beige effluent from PRO-W pond Dark effluent from control pond

20. Probiotics - Immunostimulation Improved bacterial clearance • Peptidoglycan: a component strengthening bacterial cell wall, esp. Gram(+) bacteria • peptidoglycan added to P. monodon feed resulted in • a better capacity to clear bacteria from hemolymph • better resistance to Vibrioharveyi • Boonyaratpalin et l. 2000. J. Sci. Technol. 22:633-639 • Purivirojkul et al. 2006. Kasetsart J. (Nat. Sci.) 40:181-187 • Also supplementation of feed with peptidoglycan extracted enhanced stress tolerance . • Itami et al. 1993. Proceedings of the Second Symposium on Diseases in Asian Aquaculture 0 15 30 Time after injection with Vibrioharveyi(minutes) Clearance of Vibrio from haemolymph of shrimp from control and 2 probiotic treatments (treat 1 andtreat 2). Probiotics are commercial Bacillus strains. Test carried out at SBBU, Bangkok, Thailand.

21. Probiotics - Mode of delivery • At the farm: In water in feed • At the feedmill

22. Applications Sanolife PRO-W 200g/ha (1m depth) twice in the pond preparation Sanolife PRO-W 200g/ha (1m depth) once every 15 days. Increase frequency when organic load high. Sanolife PRO-2 at 5g/Kg feed, 2 feeding/day Sanolife PRO-F at 1g/Kg Increased dosage in case of white faeces, up to 10g/Kg feed for 10 days.

23. Sanolife Nutrilake Natural sodium nitrate with silicates and trace elements • Natural fertilizer • Direct food for algae • Favors growth of diatoms and green algae • Strong Oxygenator • Oxidizes harmful reduced compounds • Healthy bio-regulator • Stimulates growth of beneficial denitrifying bacteria Sanolife Nutrilake keeps the pond bottom oxidized

24. Thank you