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

• 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

INTRODUCTION

Antibiotics

Non-absorbable chemicals

Acidifiers

Prebiotics

Probiotics

Phytogenic additives

Antibodies

Other biological substances

CONTROL & PREVENTION

• 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)

PROBLEM STATEMENT

• 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.

OBJECTIVE

Salmonella enteritidis (SE) (Salmonella enterica serovar enteritidis)

Salmonella typhimurium (ST) (Salmonella enterica serovar typhimurium)

MATERIAL & METHODOLOGY

IN-WATER TREATMENT

SE Control

Control (sterile tap water and SE)

500ml

Autoclaved tap water

Items

Homogenized

9ml 1ml overnight culture

10ml mixture

2hr 4hr 6hr 12hr 24hr

100ul

Experiment 1 : In-water Treatment

10-fold dilutions

20ul

24hours incubation 37OC

counting

100ul

Vortex mixing

plating

900ul deionized distilled water

0 2 4 6 12 240123456789

10

SE Control

hours

log1

0 cf

u/m

lResults: SE survival in Sterile - Water

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

Acidifier propionic, lactic acid & salts 7.5ml / L pH 3.5

Digestor Broiler

Lactic, citric, fumaric, activated ascorbic acid (citrex) 0.5 ml / L pH 5.9

VinegarHouse vinegar

(acetic acid)3 ml / L pH 6

SEControl

Control (sterile tap water and salmonella)

pH 7

Acidifier-treated water

500ml

Autoclaved tap water

Items

Homogenized

9ml 1ml overnight culture

10ml mixture

2hr 4hr 6hr 12hr 24hr

100ul

Experiment 1 : In-water Treatment

10-fold dilutions

20ul

24hours incubation 37OC

counting

100ul

Vortex mixing

plating

900ul deionized distilled water

0 2 4 6 12 240

2

4

6

8

10

Acidifier D. Broiler Vinager SE Control

hours

log1

0 cf

u/m

lResults: SE survival in acidified-water

ST ControlControl

(sterile tap water and ST)

In-water Treatment ST

0 2 4 6 12 240

1

2

3

4

5

6

7

8

9

ST Control hours

log

10cf

u/m

lResults: ST survival in water

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

PRODUCT INGREDIENTS DOSAGEpH of solution

in water

Acidifierpropionic, lactic acid

& salts7.5ml / L pH 3.5

Digestor Broiler

Lactic, citric, fumaric, activated ascorbic acid (citrex) 0.5 ml / L pH 5.9

VinegarHouse vinegar

(acetic acid)3 ml / L pH 6

SEControl

Control (sterile tap water and salmonella)

pH 7

Acidifier-treated water

0 2 4 6 12 240

1

2

3

4

5

6

7

8

9

Acidifier D. Broiler Vinager ST Control

hour

log1

0cfu

/ml

Results: ST survival in acidified water

• Current methodology • High survival• Drinking water• Indoor environment

• Suitable for efficacy studies…

• Digester Broiler showed remarkable

reduction of SE and ST

Discussion

In-water - a workable in vitro methodology was found.

- Digester Broiler Liquid Dosage 0.5ml / 1L water

Reduced significantly SE & ST titers.

Conclusion

In-water

….in vivo studies

under control (experimental) and/or field condition

Ultimate assessment…

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…

Acknowledgement