1. EFFECT OF INDOOR ENVIRONMENT ON THE SURVIVAL OF SALMONELLA ENTERITIDIS AND SALMONELLA TYPHIMURIUM K. Ganapathy & Kok H. S. Faculty of Veterinary Medicine University Putra Malaysia

2. INTRODUCTION • Protect health and safety of human. • Protect export market & global acceptance. • Protect health, production and welfare of chickens. • Salmonella spp infection in chickens • Prevention/control and eradication of Salmonellae infection - important

3. CONTROL & PREVENTION Antibiotics Non-absorbable chemicals Acidifiers Prebiotics Probiotics Phytogenicadditives Antibodies Other biological substances

4. PROBLEM STATEMENT • Numerous acidifiers brands for water treatment • Comparative efficacy studies – no ‘standard’ method • Efficacy of acidifiers under tropical climate is unknown • Limitation of in-vivo studies (cost, facilities, repeatability and etc)

7. IN-WATER TREATMENT

8. Items Homogenized 1ml overnight culture 9ml 500ml Autoclaved tap water 10ml mixture 2hr 4hr 6hr 12hr 24hr 100ul Experiment 1 : In-water Treatment

9. 10-fold dilutions 900ul deionized distilled water Vortex mixing 100ul plating 20ul 24hours incubation 37OC counting

10. Results: SE survival in Sterile - Water

11. NOTE Out of 7 acidifiers and/or prebiotics tested at the dosage recommended by manufactures only two products worked and had effect against the Salmonella enteritidis (SE). The acidifiers were made from different mixtures of acids such as formic, lactic, propionic, acetic, citric,. The only acidifier that worked, the problem was that it reduced so much pH of water that birds will not drink it.

12. Acidifier-treated water

13. Items Homogenized 1ml overnight culture 9ml 500ml Autoclaved tap water 10ml mixture 2hr 4hr 6hr 12hr 24hr 100ul Experiment 1 : In-water Treatment

14. 10-fold dilutions 900ul deionized distilled water Vortex mixing 100ul plating 20ul 24hours incubation 37OC counting

15. Results: SE survival in acidified-water

16. In-water Treatment ST

17. Results: ST survival in water

18. NOTE Out of 7 acidifiers and/or prebiotics tested at the dosage recommended by manufactures only two products worked and had effect against the Salmonella typhimurium (ST). The acidifiers were made from different mixtures of acids such as formic, lactic, propionic, acetic, citric,. The only acidifier that worked, the problem was that it reduced so much pH of water that birds will not drink it.

19. Acidifier-treated water

20. Results: ST survival in acidified water

22. High survival

23. Drinking water

24. Indoor environment

25. Suitable for efficacy studies…

26. Digester Broiler showed remarkable reduction of SE and ST

27. Conclusion In-water - a workable in vitro methodology was found. - Digester Broiler Liquid Dosage 0.5ml / 1L water Reduced significantly SE & ST titers.

28. Ultimate assessment… In-water ….in vivo studies under control (experimental) and/or field condition

29. Acknowledgement Research students & assistants - Dr.Mahadevan - Mr. Phi Lip - Mr. Tan Chin Giap Prof. Ho & Ms.Vicky (IBS) Prof MadyaDr.Saleha, En.Kamar, En.Hajar Pharmaceutical companies including Rhonema (M) Sdn. Bhd. Other directly or indirectly assisted…