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
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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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