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Isolation, Identification and Characterization of Salmonella from Duck

T. Mondal
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh.

M. S. R. Khan
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh.

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

M. Purakayastha
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh.

M. Das
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh.

M. P. Siddique
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh.

Abstract/ Executive Summary:

The present study was performed with the aim to isolate and identify Salmonella organism from diarrhoeic and apparently healthy ducks and to characterize duck Salmonella by biochemical test. Antibiotic sensitivity analysis of duck Salmonella was performed. A total of 65 cloacal samples were collected from ducks of three different regions such as Char Nilokkhiya, BAU Poultry Farm and Boyra, Mymensingh. Out of 65 samples 9 (13.07% ) were found positive. The antibiotic sensitivity pattern showed that the duck isolates were highly sensitive to ciprofloxacin, kanamycin, nalidixic acid, co-trimoxazole and cephalexin but these isolates were highly resistant to chloramphenicol.

Keywords: Salmonella, Duck, Isolation, Identification, Characterization

Location: Bacteriology laboratory of the Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University (BAU), Mymensingh.

Start Date: 00-11-2006

End Date: 00-10-2007

Program Area: Pest Management

Commodity: Duck

Objectives:

To isolate and identify Salmonella organism from diarrhoeic and apparently healthy ducks and to characterize duck Salmonella by biochemical test.

Methodology:

The present research was conducted between November 2006 to October 2007 in the Bacteriology laboratory of the Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University (BAU), Mymensingh. A total of 65 cloacal swab samples, 24 from Bangladesh Agricultural University (BAU) Poultry Farm, 22 from Boyra, Mymensingh and 19 from Char Nilokkhiya were used in this study. The samples were collected from 40 apparently healthy ducks and 25 diarrhoeic ducks for bacteriological study. The ducks were the breeds of Khaki Campbell and Indian Runner and age ranged from 2 to 3 months without considering any sex. Sterile cotton swab and sterile Bacto- selenite broth in sterile glass tubes were carried to the sampling areas and samples were collected from the mucosa of the cloaca of the ducks. The swabs collected aseptically were transferred immediately into Bacto selenite broth and test tubes containing swab samples were then immediately brought to the bacteriology laboratory, Department of Microbiology and Hygiene, BAU and subjected to cultural study . Each of the collected cloacal swabs was inoculated into freshly prepared selenite broth. Then the tubes were marked properly and incubated at 37ºC for 24 hours aerobically in bacteriological incubator. The incubated tubes were then examined for growth of bacteria. 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, Salmonella-Shigella agar and Brilliant green agar separately. The plates were then incubated at 37ºC for 24 hours and the plates containing characteristic colonies of Salmonella are selected. Motility test and Gram’s staining test are performed to identify the plates containing Salmonella accurately. Subculturing in Salmonella -Shigella agar was performed from the suspected plates containing Salmonella to obtain a pure culture. These pure isolates obtained in this way were used for further study. A small colony from the representative Salmonella colonies was picked up from SS, MC and BGA plates with a bacteriological loop, smeared on separate glass slide and fixed by gentle heating. Crystal violet was then applied on each smear to stain for two minutes and then washed with running water. Few drops of Gram’s iodine was then added to act as mordent for one minute and then again washed with running tap water. Acetone alcohol was then add ed (acts as decolorizer) for few seconds. After washing with water, safranin was ad ded as counter stain and allowed to stain for 2 minutes. Then the slides were washed with water, blotted and dried in air and then examined under microscope with high power objective (100X) using immersion oil. The motility test was performed to differentiate motile bacteria from non-motile one. The motile and non-motile organisms were identified by ob serving motility in contra sting with swinging movement of bacteria. The motile bacteria with swinging movement were identified as Salmonella. Isolated organisms with supporting growth characteristics of Salmonella on various media were maintained on SS and BGA and were subjected to the following biochemical tests named sugar fermentation test, MR-VP reaction and indole reaction. The carbohydrate fermentation test was performed by inoculating a loopful of thick bacterial culture into the tubes containing five basic sugars (dextrose, malt ose, sucrose, lactose, and mannitol) and incubated at 37ºC for 24 hours. The Methyl test, Voges-Proskauer test and Indole test was conducted. Susceptibility of the isolated Salmonellae to different antibacterial agents was performed through disc diffusion method to determine the drug sensitivity pattern. The antibacterial discs used were erythromycin, amoxicillin, cephalexin, chloramphenicol, co-trimoxazole, kanamycin, ciprofloxacin and nalidixic acid. Salmonella isolates were grown overnight on BGA and the overnight cultured isolates were inoculated into NB and poured on BGA and spreaded 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 at 37ºC for 24 hours, aerobically.

Source of Information: Bangl. J. Vet. Med. (2008), 6 (1): 07–12

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Achievement/Findings:

From the antibiogram study, it was revealed that the isolates were highly sensitive to ciprofloxacin, kanamycin, nalidixic acid, and cephalexin while moderately sensitive to kanamycin, nalidixic acid, co-trimoxazole, cephalexin, erythromycin and amoxicillin. The isolates were less sensitive to erythromycin, amoxicillin and chloramphenicol whereas they were resistant to chloramphenicol.

Knowledge Management: Journal