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Isolation, Identification and Characterization of Bacterial Flora from the Respiratory Tract of Apparently Healthy Sheep

Chandan Kumar Sarker
Veterinary Surgeon, Department of Livestock Services, Farmgate, Dhaka, Bangladesh

Anowar Hossen
Poultry Disease Diagnostic Laboratory, Poultry Production Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka, Bangladesh

Md. Abu Yousuf
Scientific Officer
SAARC Regional Leading Diagnostic Laboratory for PPR, Animal Health Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka-1341, Bangladesh.

Md. Ala Uddin
Department of Animal Husbandry and Veterinary science, University of Rajshahi, Rajshahi, Bangladesh

Most. Sumona Akter
Veterinary Surgeon, Department of Livestock Services, Farmgate, Dhaka, Bangladesh

Mostafizur Rahman
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh, Bangladesh

Md. Bahanur Rahman
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh, Bangladesh

Abstract/ Executive Summary:

Bangladesh is a densely populated developing country and its economy is primarily based on agriculture. The current study was designed for isolation, identification and characterization of bacterial flora from the upper respiratory tract of sheep. Thirty apparently healthy sheep were selected at the adjacent areas of Bangladesh Agricultural University (BAU) for this experiment. Swab samples were collected from nasal swabs, lung swabs and tracheal swabs. All samples were subjected into inoculated on to bacteriological media . Furthermore, all of the bacterial isolates were characterized by Gram’s staining, biochemical tests, antibiotics sensitivity tests and pathogenicity tests. None of the isolated Bacillus, E. coli and Staphylococcus spp. was found to be pathogenic. Isolated Pasteurella spp. were found to be pathogenic as observed in different experimental models and showed a degree of variation in antibiotic drug sensitivity test. Ciprofloxacillin was sensitive to all of the isolated bacteria. Through the bacteria that were isolated from various organs of apparently healthy sheep is normal micro flora, however these may act as primary pathogen and may produce diseases when the sheep are immunologically suppressed due to severe stress conditions.

Keywords: Sheep, Respiratory tract, Bacteria, Antibiotics

Location: Bangladesh Agricultural University, Mymensingh, Bangladesh

Start Date:

End Date:

Program Area: Animal Health and Management

Commodity: Sheep, Diseases

Objectives:

Objective of this study was to reduce the mortality rate of sheep from bacterial diseases, minimize cost of healthy sheep production, reduce economic loss which is spent for the treatment of sheep diseases resulting more establishments of sheep farms that creates employment opportunity and will increase animal protein production.

Methodology:

Thirty apparently healthy sheep were selected at the adjacent areas of Bangladesh Agricultural University (BAU) for this experiment and under the Department of Microbiology and Hygiene. Swab samples were collected from Nasal swabs, Lung swabs and Tracheal swabs. Nasal swabs, tracheal swabs and lungs swabs were collected using sterilized cotton with aseptically. Then the swabs were inoculated into NB and selenite broth and incubated at 37⁰C for 24 hours. After incubation it was streaked on to NA and MC agar medium. Bacterial pathogen free healthy day-old Swiss Albino suckling mice were selected for this experiment. All the mice were supplied by the laboratory experimental animal house of the Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh. After inoculation of the inoculum (bacteria, crude toxin and both bacteria and crude toxin) the laboratory animals were kept in an isolation room of the animal shed of the Department of Microbiology and Hygiene providing appropriate temperature. The entire study was divided into three major steps: The first step included selection of sources, collection of samples, isolation, identification and characterization of isolates organisms on the basis of their colony morphology, staining property, motility and biochemical characteristics. In the second step, pathogenicity study of the isolates organisms was performed in different experimental model such as in day-old chicks and in day-old suckling mice to observe their lethal effect. In the third step, the current status of drug sensitivity and resistance pattern of the bacterial isolates from nasal, tracheal and lung swabs was determined. Bacteriological media: a. Agar media; b. Liquid media (broth); c. Chemicals and reagents. To determine the drug sensitivity and resistance pattern and to interpret their disease potential commercially available antimicrobial discs (Becton, Dickinson and Company, USA) were used. This method allowed for the rapid detection of the efficacy of drugs against the test organisms by measuring the diameter of the zone of inhibition that resulted from diffusion of the agent into the medium surrounding the discs inhibiting the growth of the organisms. The following antimicrobial agents with their disc concentration were used to test the sensitivity and resistance pattern of the selected bacterial isolates from nasal, tracheal and lung swab of sheep. Isolation and identification of Pasteurella spp., Isolation and identification of Bacillus spp. Gram’s staining was performed as per recommendation of Merchant and Packer (1976) to determine the size, shape and arrangement of bacteria. The procedure was as follows: A small colony was picked up with a bacteriological loop, smeared on a glass slide and fixed by gentle heating. Crystal violet solution was then applied on the smear to stain for two minutes and then washed through running tap water. Gram’s iodine was then added to act as a mordent for one minute and then again washed with running water. Acetone alcohol was then added, which act as a decolorizer. After washing with water, safranine was added as counter stain and allowed to stain for 2 minutes. The slide was then washed with water, blotted and air dried and then examined under microscope with high power objective (100X) using immersion oil.

Source of Information: Asian J. Med. Biol. Res. 2015, 1 (3), 677-685; ISSN 2411-4472

Article Link (Online Journal): http://banglajol.info/index.php/AJMBR/article/view/26496/17787

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

From the present study it may be concluded that among all the isolates the percentage of the Pasteurella is higher than Bacillus. None of the isolated Bacillus was found to be pathogenic. Isolated Pasteurella spp. were found to be pathogenic as observed in different experimental models and showed a degree of variation in antibiotic drug sensitivity test. Ciprofloxacillin were sensitive to all of the isolated bacteria. Through the bacteria that were isolated from various organ of apparently healthy sheep are normal micro flora, however these may act as primary pathogen and may produce diseases when the sheep are immunologically suppressed due to severe stress conditions.

Knowledge Management: Journal