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Isolation and Detection of Antibiotic Sensitivity Pattern of Escherichia Coli from Ducks in Bangladesh and Nepal

Avishek Singh
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Md. Shahidur Rahman Khan*
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Sukumar Saha
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Jayedul Hassan
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Urmi Roy
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh

Abstract/ Executive Summary:

A total of 120 rectal swab samples from ducks (sixty samples from Nepalgunj area of Nepal and 60 from Boyera area under Mymensingh district of Bangladesh) were collected for the isolation of Escherichia coli and their antibiogram study. After cultural and biochemical examination, a total of 40 samples from Nepalgunj and 45 samples from Boyera were found positive for E. coli. Pathogenicity study of 10 positive isolates from Nepalgunj and 12 positive isolates from Boyera were done to detect the presence of enterotoxin. All inoculated mice died showing typical lesion of extensive hemorrhage and massive edema. The isolates from two different origins showed major difference in their antibiogram study. The isolates of Nepal were highly sensitive to ciprofloxacin, co-trimoxazole, chloramphenicol and amoxicillin; moderately sensitive to nalidixic acid and; less sensitive to kanamycin and resistant to cephalexin. However, the isolates of Bangladesh were highly sensitive to ciprofloxacin, chloramphenicol and amoxicillin; moderately sensitive to nalidixic acid, cephalexin, and co-trimoxazole; less sensitive to kanamycin. This variation of antibiotic sensitivity and resistance patterns among the E. coli isolates of Nepal and Bangladesh might be due to strain variations and indiscriminate use of antibiotics in these two different countries.

Keywords: Escherichia coli, Duck, Nepal, Bangladesh, Antibiogram

Location: Department of Microbiology and Hygiene, Bangladesh Agricultural University (BAU), Mymensingh-2202, Bangladesh.

Start Date: 00-07-2002

End Date: 00-11-2002

Program Area: Pest Management

Commodity: Duck

Objectives:

This study was carried out to characterize the E. coli isolates from duck of Bangladesh and Nepal and to find out the effective antibiotic(s) that could be applied to control avian colibacillosis in duck in these areas.

Methodology:

The study was conducted during July 2010 to Nov. 2010 at the Department of Microbiology and Hygiene, Bangladesh Agricultural University (BAU), Mymensingh-2202, Bangladesh. A total of 60 field samples (rectal swabs) were collected from 35 diarrheic and 25 apparently healthy ducks by sterile cotton bud and carried to the laboratory of Nepalgunj Medical College and Hospital, Nepalgunj, Nepal. All these 60 samples were inoculated on Eosine Methylene Blue (EMB) agar plates, incubated for overnight at 37⁰C. The suspected colonies were brought to the Laboratory of Bacteriology, Department of Microbiology and Hygiene, at BAU, Mymensingh maintaining cool chain. Similarly, 60 field samples comprising rectal swab were collected by sterile cotton bud from 35 diarrheic and 25 apparently healthy ducks of Boyera area, located around BAU, Mymensingh, Bangladesh and brought to laboratory for isolation and characterization of E. coli. Isolation and characterization of isolated E. coli Pure culture was obtained from the primary culture using EMB agar, MacConkey (MC) agar, Brilliant Green (BG) agar, Salmonella-Shigella (SS) agar and Blood agar. The colony characteristics were observed and Gram staining was performed. Motility test and biochemical tests (Sugar Fermentation test, Indole test, Methyl-Red (MR) test, Voges-Proskauer (VP) test and Catalase test) were performed for further characterization of the isolates of both origins. Pathogenicity study of the isolates: A total of 10 positive isolates from Nepalgunj and 12 positive isolates from Boyera were subjected for pathogenicity study. A total of 30 new born Swiss Albino suckling mice (2-4 days old) collected from the Laboratory Animal Center of the Department of Microbiology and Hygiene, BAU were divided into two groups (n=15/ group): group A for isolates from Nepalgunj and group B for isolates from Boyera on random basis. Ten EMB colonies of isolates from Nepalgunj and 12 colonies of isolates from Boyra were inoculated into nutrient broth and incubated overnight at 37⁰C. The cells were then removed from the broth culture by centrifugation at 25,000 rpm for 20 min. The supernatants were filtered and collected. Each of the 12 mice in group A received 0.1 ml of supernatant by oral route and remaining 3 mice were kept as control. Similarly, 10 mice from group B were administered 0.1 ml of supernatant and 5 mice from this group were kept as control without enterotoxin treatment. Both groups were kept at room temperature for 14 hours. After 14 hours all mice (live and dead) including the control mice, were necropsied. The ratio of gut weight and the remaining carcass weight was calculated for each mouse and the average ratio was calculated (Gyles, 1979). Antibiogram of isolates: Eight different antibacterial discs (HiMedia, Mumbai, India) were selected randomly for antibiotic sensitivity study against isolated E. coli from Bangladesh and Nepal (Table 1). Six isolates from each of the country were subjected for antibiogram study. A disk diffusion test of the Kirby-Bauer (Bauer et al., 1966) method was used to determine the susceptibility of E. coli isolates to different antibiotic agents. The procedure involved measuring the diameter of the zone of inhibition that results from diffusion of the agent into the medium surrounding the disc. Tentative inhibition zone diameter interpretive criteria were developed using the error-rate-bounded methods recommended by the CLSI (Clinical and Laboratory Standards Institute, 2011)

Source of Information: Microbes and Health, June 2012, 1(1): 6-8 ISSN: 2226-0153 (Print)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Based on this study, it may be concluded that chloramphenicol, ciprofloxacin and amoxicillin should be the first choice of treatment against avian colibacillosis in duck in Bangladesh. On the other hand amoxicillin, co-trimoxazole, ciprofloxacin and chloramphenicol should be the first choice of drug for the treatment of avian colibacillosis in duck in Nepal. The resistance pattern observed among the isolated E. coli in Bangladesh and Nepal might be related to the extensive and indiscriminate use of antibiotic in duck in these areas.

Knowledge Management: Journal