The study tested the efficacy of 7 acidifiers and prebiotics in reducing Salmonella enteritidis (SE) and Salmonella typhimurium (ST) in drinking water under indoor environmental conditions in Malaysia. Only two products were effective against SE and ST: Digester Broiler Liquid at a dosage of 0.5ml per liter of water remarkably reduced levels of both pathogens. While another acidifier reduced pathogen levels, it lowered the water pH too much for chickens to drink. In-vitro methodology proved suitable for efficacy studies, but in-vivo studies under controlled or field conditions are still needed for ultimate assessment.

1. Effect of indoor environment on the survival of S almonella 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.

3. Introduction • Salmonella spp infection in chickens • Prevention/control and eradication of Salmonellae infection - important

6. Objective  To determine survival of SE and ST in the drinking water under Malaysian indoor environment.  To determine survival of SE and ST in of 7 different products in water under Malaysian indoor environment.

7. Material & Methodology  Salmonella enteritidis (SE) ( Salmonella enterica serovar e nteritidis )  Salmonella typhimurium (ST) ( Salmonella enterica serovar typhimurium )

8. In-water Treatment Control (sterile tap water and SE) SE Control

9. Experiment 1 : In-water Treatment 500ml autoclaved tap water items homogenized 9ml 1ml overnight culture 10ml mixture 2hr 4hr 6hr 12hr 24hr 100 u l

10. 10- fold dilutions 900 u l deionized distilled water 20 u l 24hours incubation 37 O C counting 100 u l Vortex mixing plating

11. Results: SE survival in sterile-water

12. 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. NOTE

13. Acidifier-treated water 3 ml / L 0.5 ml / L 7.5ml / L Control (sterile tap water and salmonella) House vinegar (acetic acid) Lactic, citric, fumaric, activated ascorbic acid (citrex) propionic, lactic acid & salts pH 7 pH 6 pH 5.9 pH 3.5 Vinegar SE Control Digestor Broiler Acidifier

14. Experiment 1 : In-water Treatment 2hr 4hr 6hr 12hr 24hr 100 u l 500ml autoclaved tap water items homogenized 9ml 1ml overnight culture 10ml mixture

15. 10- fold dilutions 900 u l deionized distilled water 20 u l 24hours incubation 37 O C counting 100 u l Vortex mixing plating

16. Results: SE survival in acidified-water

17. In-water Treatment ST Control (sterile tap water and ST) ST Control

18. Results: ST survival in water

19. 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. NOTE

20. Acidifier-treated water pH of solution in water DOSAGE INGREDIENTS PRODUCT 3 ml / L 0.5 ml / L 7.5ml / L Control (sterile tap water and salmonella) House vinegar (acetic acid) Lactic, citric, fumaric, activated ascorbic acid (citrex) propionic, lactic acid & salts pH 7 pH 6 pH 5.9 pH 3.5 Vinegar SE Control Digestor Broiler Acidifier

21. Results: ST survival in acidified water

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

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

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