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Prevalence and Characterization of Escherichia Coli from Rectal Swab of Apparently Healthy Cattle in Mymensingh, Bangladesh

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

Md. Bahanur Rahman
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

Tazrin Kamal
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

K. H. M. Nazmul Hussain Nazir
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

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

Md. Shafiullah Parvej
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

Prevalence study of commensal E. coli in the rectal swab of apparently healthy animals is essential to reveal out the epidemiology of disease outbreaks and development of antibiotic resistance by E. coli. Though there are reports on the prevalence of pathogenic E. coli, information on the commensal E. coli are very few in Bangladesh. This study will provide a base line data on the prevalence of E. coli in the rectal swab of different groups of cattle in the study area. However, the area selected and sample examined in this study is very few, a study comprising larger population size origination from other part of the country will reveal the actual figure of prevalence E. coli in the rectal Swab of apparently healthy cattle in Bangladesh.

Keywords: Prevalence, Characterization, E. coli, Rectal swab, Apparently healthy cattle

Location: Dairy Farm, Bangladesh Agricultural University (BAU), Bangladesh

Start Date:

End Date:

Program Area: Animal Health and Management

Commodity: Cattle

Objectives:

1. The present study was designed to investigate the prevalence of E. coli in the rectal swab of apparently healthy cattle reared in Bangladesh Agricultural University (BAU) dairy farm and visiting BAU veterinary clinic, Mymensingh, Bangladesh.

Methodology:

Sample collection - The rectal swab samples (n=128) were collected randomly from apparently healthy cattle comprising 35 samples from the Veterinary Clinic, and 93 samples from Dairy Farm, Bangladesh Agricultural University (BAU), Bangladesh following a convenience sampling method without repetition of animals. Sterile cotton buds were used for the collection of swab samples, and the swab was transferred to nutrient broth instantly. The swab samples were transported to the Bacteriology Laboratory at the Department of Microbiology and Hygiene, BAU. During sample collection information regarding the age, sex, breed and management systems of the animals were also recorded. Cultural and biochemical examination - The nutrient broth containing swab samples were incubated overnight at 37^oC. After overnight incubation samples from the nutrient broth were cultured on to EMB agar, MacConkey agar and Cefixime Tellurite – Sorbitol MacConkey (CT-SMAC) medium. Isolated organisms with supporting growth characteristics of E. coli were subjected to sugar (dextrose, fructose, maltose, lactose and sucrose) fermentation, MR-VP and indole production test. DNA Extraction - Crude DNA was obtained from the isolates using boiling method with little modification. Briefly, the organisms were cultured onto EMB agar at 37^oC. After overnight incubation a medium sized colony was picked up with sterile tips and mixed in 200μl of deionized water. The mixture was then heated in boiling water for 10 minutes followed by dipping into ice for 10 minutes and centrifugation at 10,000rpm for 10 minutes. The supernatant was collected which contains DNA. The DNA sample was kept in -20^oC until use. PCR for the confirmation of the isolates as E. coli - The isolated organisms that were preliminarily identified as E. coli were confirmed by PCR using primers specific to E. coli 16S rRNA gene. PCR was performed with slight modification. 25μl reaction containing 1X Taq Polymerase PCR master mix and 10pmol of primer was prepared. After initial incubation at 95^oC for 3 min, a 30-cycle amplification protocol was followed as 94^oC for 45 s, 58^oC for 45 s and 72^oC for 60 s, and a final extension step of 72^oC for 3 min. Electrophoresis of the PCR products was done using 2% agarose gel. After electrophoresis, the gel was stained for 10 minutes in ethidium bromide for visualization. Data analysis - Data were entered and statistical analyses were performed using SPSS version 17. Using Pearson’s Chi-square (χ2) test difference among the variables was calculated. P values less than 0.05 were considered as significant.

Source of Information: Microbes and Health, June 2014, 3(1): 12-14; ISSN: 2226-0153 (Print) 2305-3542 (Online)

Article Link (Online Journal): http://journal.bsvmph.org/

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Prevalence study of commensal E. coli in the rectal swab of apparently healthy animals is essential to reveal out the epidemiology of disease outbreaks and development of antibiotic resistance by E. coli. Though there are reports on the prevalence of pathogenic E. coli, information on the commensal E. coli are very few in Bangladesh. This study will provide a base line data on the prevalence of E. coli in the rectal swab of different groups of cattle in the study area. However, the area selected and sample examined in this study is very few, a study comprising larger population size origination from other part of the country will reveal the actual figure of prevalence E. coli in the rectal Swab of apparently healthy cattle in Bangladesh.

Knowledge Management: Journal