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Molecular Characterization of Salmonella Isolates of Duck in Comparison to Salmonella Isolates of Chicken and Ruminants

Tania Mondal
Department of Microbiology & Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh,

M Shahidur Rahman Khan
Department of Microbiology & Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh,

Munirul Alam
Enteric Microbiology Laboratory, Laboratory Sciences Division, Centre for Health & Population Research, International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B), GPO Box 128, Mohakhali, Dhaka 1212, Bangladesh.

Moushumi Purakayastha
Department of Microbiology & Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh,

Abstract/ Executive Summary:

Salmonella species are recognized as a major cause of food borne illness that are closely associated with the consumption of contaminated poultry and egg products. The present study was conducted to compare the cultural, biochemical characteristics, antibiotic sensitivity pattern and the patterns of genomic organization of duck Salmonella isolates associated with chicken, cattle, sheep and goat using pulsed-field gel electrophoresis (PFGE) with Xba1 restriction enzyme. The comparative antibiogram study among duck, chicken and ruminants showed variable results in antibiotic sensitivity and similar results in resistance pattern. Genome analysis using PFGE with Xba1 restriction enzyme revealed that the Salmonella isolates of the same species collected from same areas to be of same genomic pattern, although a great genomic diversity could be found among duck, chicken, sheep, goat and cattle Salmonella strains. It may be concluded from the result of this research work that the heterogeneity in genomic organization among different isolates of different species collected from different areas occurred greatly and for this reason.

Keywords: Salmonellosis, Salmonella, Pulsed-field gel electrophoresis, Duck, Chicken, Ruminants

Location: BAU, Mymensingh

Start Date: 00-11-2006

End Date: 00-10-2007

Program Area: Variety and Species

Commodity: Chicken

Objectives:

To compare the cultural, biochemical characteristics, antibiotic sensitivity pattern and the patterns of genomic organization of duck Salmonella isolates associated with chicken, cattle, sheep and goat using pulsed-field gel electrophoresis (PFGE) with Xba1 restriction enzyme.

Methodology:

The experiment was conducted between November 2006 and October 2007 in the Bacteriology Laboratory of the Department of Microbiology and Hygiene, Bangladesh Agricultural University(BAU), Mymensingh and Enteric Microbiology Laboratory, International Centre for Diarrhoeal Diseases Research of Bangladesh (ICDDR,B) in Dhaka. A total of 65 cloacal swab samples from apparently healthy and diarrhoeic ducks were tested for the examination. In addition, 27 isolates of Salmonella from chicken, cattle, sheep and goat collected from the repository of the Department of Microbiology and Hygiene, BAU, Mymensingh were used to compare with those of ducks Cloacal swabs were collected and each of swabs was inoculated into freshly prepared selenite broth. Then the tubes were marked properly and incubated at 37ºC for 24 h aerobically in bacteriological incubator. The tubes were then examined for growth of bacteria. Smears were prepared for each culture and the Gram-stained examined under microscope. Gram-negative rod isolates were streaked on MacConkey agar, Salmonella-Shigella agar and brilliant green agar separately. The plates were then incubated at 37ºC for 24 h. The plates containing characteristic colonies of Salmonella were selected. Motility test and Gramreaction were performed to identify Salmonella. Sub culturing in Salmonella-Shigella agar was performed from the suspected plates containing Salmonella to obtain a pure culture¹². These pure isolates obtained in this way were used for further study Organisms showing cultural characteristics of Salmonella on various media were maintained on SS and BGA and were subjected to biochemical tests such as sugar fermentation test, MR-VP reaction and indole reactionSusceptibility of the Salmonella isolates to different antibacterial agents was performed through disc diffusion method13. In this method Salmonella isolates were grown overnight on BGA. The overnight cultured isolates were inoculated into NB and poured on BGA and spread uniformly with the help of sterile glass spreader. Antibacterial discs were applied aseptically to the surface of the plate at an appropriate arrangement with the help of sterile forceps and incubated aerobically at 37°C for 24 h¹⁴Salmonella isolates from duck and the laboratory isolates of Salmonella from chicken, cattle, sheep and goat were preserved in 20% glycerine and soft agar method. Salmonella isolates preserved in 20% glycerine were placed in ice box and transported to ICDDR,B, Dhaka for performing molecular characterization Molecular typing of Salmonella by pulsed-field gel electrophoresis (PFGE) Molecular typing of Salmonella isolates by PFGE was done using preparation of PFGE agarose plugs from cell suspensions and lysis of cells in agarose plugs. After cell lysis agarose plugs were washed and then subjected to restriction digestion of DNA in agarose plugs with Xba1 followed by casting agarose gel and loading restriction plug slices on the comb. Electrophoresis was performed with the contour clamped homogenous electric field (CHEF-DRII) apparatus from the Bio-Rad (Richmond, USA ).

Source of Information: Bangladesh J Microbiol, Volume 25, Number 2, December 2008, pp 91-94

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The comparative antibiogram study among duck, chicken and ruminants showed variable results in antibiotic sensitivity and similar results in resistance pattern. Genome analysis using PFGE with Xba1 restriction enzyme revealed that the Salmonella isolates of the same species collected from same areas to be of same genomic pattern, although a great genomic diversity could be found among duck, chicken, sheep, goat and cattle Salmonella strains. It may be concluded from the result of this research work that the heterogeneity in genomic organization among different isolates of different species collected from different areas occurred greatly and for this reason.

Knowledge Management: Journal