5 Salmonella Water Malaysia

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K. Ganapathy & Kok H. S.Faculty of Veterinary Medicine

University Putra Malaysia

Effect of indoor environment on the survival of

Salmonella enteritidis and

Salmonella typhimurium

Introduction

• Protect health and safety of

human.

• Protect export market & global

acceptance.

• Protect health, production and

welfare of chickens.

Introduction

• Salmonella spp infection in

chickens

• Prevention/control and eradication

of Salmonellae infection - important

Control & PreventionAntibiotics

Non-absorbable chemicals

AcidifiersPrebioticsProbioticsPhytogenic additivesAntibodies

Other biological substances

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)

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.

Material & Methodology

Salmonella enteritidis (SE) (Salmonella enterica serovar enteritidis)

Salmonella typhimurium (ST) (Salmonella enterica serovar typhimurium)

In-water Treatment

SE Control

Control

(sterile tap water and SE)

Experiment 1 : In-water Treatment

500ml

autoclaved tap water

items

homogenized

9ml1ml overnight culture

10ml mixture

2hr 4hr 6hr 12hr 24hr

100ul

10-fold dilutions

900ul deionized distilled water

20ul

24hours incubation 37OC

counting

100ul

Vortex mixing

plating

Results: SE survival in sterile-water

0

2

4

6

8

10

0 2 4 6 12 24

hours

log1

0 cf

u/m

l

SE Control

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-treated water

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

Experiment 1 : In-water Treatment

2hr 4hr 6hr 12hr 24hr

100ul

500ml

autoclaved tap water

items

homogenized

9ml1ml overnight culture

10ml mixture

10-fold dilutions

900ul deionized distilled water

20ul

24hours incubation 37OC

counting

100ul

Vortex mixing

plating

Results: SE survival in acidified-water

0

2

4

6

8

10

0 2 4 6 12 24hours

log1

0 cf

u/m

l

Acidifier D. Broiler Vinager SE Control

In-water Treatment ST

ST Control

Control

(sterile tap water and ST)

0123456789

0 2 4 6 12 24hours

log1

0cfu

/ml

ST Control

Results: 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

Acidifier-treated water

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

Results: ST survival in acidified water

0

1

2

3

4

5

6

7

8

9

0 2 4 6 12 24hour

log1

0cfu

/ml

Acidifier D. Broiler Vinager ST Control

Discussion

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

• Suitable for efficacy studies…

• Digester Broiler showed remarkable

reduction of SE and ST

Conclusion

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

- Digester Broiler Liquid Dosage 0.5ml / 1L water

Reduced significantly SE & ST titers.

Ultimate assessment…

In-water

….in vivo studies

under control (experimental) and/or field condition

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

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