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Cultural and Biochemical Characterization of Sheep Escherichia Coli Isolated from in and Around BAU Campus

M. Purkayastha
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh

M. S. R. Khan
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh

M. Alam
Enteric Microbiology Laboratory, Laboratory Sciences Division, ICDDR,B, Dhaka,

M. P. Siddique
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh

F. Begum
Veterinary Training Institute, Khakdahor, Mymensingh

T. Mondal
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh

S. Choudhury
1Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh

Abstract/ Executive Summary:

Pathogenic Escherichia coli remain as an important etiological agent of sheep diarrhoea in Bangladesh. The present study was designed for the cultural and biochemical characterization of Sheep E. coli from diarrhoeic and apparently healthy sheep in and around BAU campus for the period from January to October, 2007. Out of 90 faecal samples, 36 from diarrhoeic and 54 from apparently healthy sheep collected from different areas in and around BAU campus, 15 (41.67%) and 21 (38.38%) were found to be positive for E. coli. The cultural characterization of all positive sheep E. coli revealed greenish black colony with metallic sheen in Eosine methylene blue agar, bright pink color smooth transparent colony in MacConkey agar, green color colony in Brilliant green agar, slight pinkish smooth colony in Salmonella-Shigella agar and colorless colony with hemolysis in blood agar. In case of biochemical characterization, all of the isolates showed fermentation of dextrose, sucrose, fructose, maltose and mannitol with the production of acid and gas, negative result to Voges-Proskaure test, positive result to Methylred test and differential result to Indole test.The overall prevalence of E. coli was recorded as 80.05% through the cultural and biochemical characterization. The antibiotic sensitivity and resistance pattern showed that the isolates of sheep E. coli were highly sensitive to ciprofloxacine, co-trimoxazol, nalidixic acid and chloramphenicol but to the erythromycin the isolates were highly resistant

Keywords: E. coli, Sheep, Biochemical, characterization

Location: Mymensingh

Start Date: 01-01-2007

End Date: 31-10-2007

Program Area: Animal Health and Management

Commodity: Sheep

Objectives:

The present study was undertaken with some distinct aims these are isolation, identification, biochemical characterization and antibiotic sensitivity pattern of E. coli isolated from sheep faeces.

Methodology:

The study was conducted in the Bacteriology laboratory of the Department of Microbiology and Hygiene, BAU, Mymensingh during the period of January 2007 to October 2007.The study was conducted in the Bacteriology laboratory of the Department of Microbiology and Hygiene, BAU, Mymensingh during the period of January 2007 to October 2007.Primary growth of all kinds of bacteria was performed in NB. Rectal swabs were collected with sterile cotton bud by gentle touch, and then inoculated into the NB, incubated for overnight at 37°C to obtain the primary culture. After primary culture of the organism smears were prepared from each of the test tubes and the smears were fixed. The fixed smears were stained with Gram’s Method of staining and examined under microscope at 100 magnifications using immersion oil. In presence of gram negative rods in the smears, the materials from the tube corresponding to the smears were streaked into MacConkey agar, Eosin-Methyline Blue agar, Salmonella- Shigella agar and Brilliant green agar separately. The plates were then incubated for overnight at 37°C and the plates containing characteristic colonies of E. coli were selected for subculture. Motility test and Gram’s staining testare performed to identify the plates containing E. coli accurately. Subculturing in Eosin - Methyline Blue agar was performed from the suspected plates containing E. coli obtain a pure culture. These pure isolates obtained in this way were used for further study. (Cheesbrough, 1984). A small colony was picked up with a bacteriological loop, smeared on a glass slide and fixed by gently heating. Crystal violet solution was then applied on the smear to stain for two minutes and then washed with running water. Lugol’s iodine was then added to act as mordant for one minute and then again washed with running water. Acetone alcohol was then added, which act as a decolorizer, for 5 seconds. After washing with water, safranine was added as counter stain and allowed to stain for two minutes. The slide was then washed with water, blotted and dried in air and then examined under microscope with high power objectives (100X) using immersion oil (Merchant and Packer, 1967).

Source of Information: Bangl. J. Vet. Med. (2010). 8(1): 51 – 55

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In the present study, out of 90 rectal swab samples, 37 samples were identified as E. coli in which 15 were collected from diarrhoeic sheep and 21 were collected from apparently healthy sheep belonging to different areas in and around BAU campus. This study revealed that the higher prevalence rate was from diarrhoeic sheep (41.67%) and the lower from apparently healthy sheep (38.88%)The cultural characterization of all positive sheep E. coli revealed greenish black colony with metallic sheen in Eosine methylene blue agar, bright pink color smooth transparent colony in MacConkey agar, green color colony in Brilliant green agar, slight pinkish smooth colony in Salmonella-Shigella agar and colorless colony with hemolysis in blood agar. In Gram’s staining, the morphology of the isolated bacteria exhibited Gram negative short rod arranged in single or paired and after motility test we also found that they were motile. In biochemical examination all the isolates fermented dextrose, sucrose, fructose, maltose and mannitol with the production of acid and gas within 24-48 hrs of incubation. The isolates also revealed positive reaction in MR test, negative reaction in VP test and differential results in Indole test. In the present study we found that the isolates of sheep E. coli was highly sensitive to ciprofloxacin, cotrimoxazol, nalidixic acid and chloramphenicol while moderately sensitive to kanamycine and amoxicillin and less sensitive to cephalexin. The isolates of sheep E. coli were fully resistant (100%) to erythromycin

Knowledge Management: Journal