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Isolation and Identification of Avibacterium Paragallinarum from Layer Chickens in Gazipur, Bangladesh

Sharmin Akter
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

Kamrul Ahmed Khan
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

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

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

Abstract/ Executive Summary:

An investigation was conducted for isolation, identification and determination of antibiotic sensitivity of Avibacterium paragallinarun, the causal agent of infectious coryza, from layer chickens. A total of 21 samples with characteristic symptoms of the disease were collected from a Hatchery of Gazipur. Tissue specimens obtained aseptically from swollen infra orbital sinus and tracheal swab were processed, of which, 3 were found positive while the rest 18 were negative. Isolation of bacteria was performed by first putting the specimen in Nicotinamide adenine dinucleotide (NAD) enriched phosphate buffer broth, anaerobically incubated for 24 hours followed by culturing loopful of broth on Blood agar (BA) and Chocolate agar (CA) plates enriched with NAD and streaked with feeder organism of Staphylococcus. aureus. On 24 hours of anaerobic incubation (candle jar method), dew drop satellite colonies of A. paragallinarum were visible on the culture plates. Cultural characteristics of bacteria as well as their staining, morphological, motility and biochemical properties such as sugar fermentation, MR and V-P tests, Indole production and catalase tests were recorded for identification. Further, antibiogram study revealed that the isolates were sensitive to Ciprofloxacin, Chloramphenicol and Gentamicin but resistant to Ampicillin, Amoxycillin, Oxytetracycline, Erythromycin and Sulphamethoxazole.

Keywords: Infectious coryza, Infraorbital swelling, Feeder organism, Antibiogram profiles

Location: Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh

Start Date: 00-06-2012

End Date: 00-07-2013

Program Area: Animal Health and Management

Commodity: Chicken

Objectives:

1. The present research work was undertaken to isolate and identify Avibacterium paragallinarum from infected layer chicken, determination of dependence on growth factor requirement to isolate Avibacterium paragallinarum and their antibiotic sensitivity profiles against commonly used antimicrobial agents.

Methodology:

A total of 21 samples (of which 15 were from live and 6 dead Chickens) were collected from Phenix Hatcheries Ltd. during the period of June 2012 to July 2013. Aseptically collected materials from swollen infraorbital sinus and tracheal swabs were used as laboratory specimens. Isolation of Bacteria - Bacteriological samples obtained from nasal, tracheal and infraorbital sinus swabs inoculated separately in glycerol-enriched phosphate buffered broth supplemented with NAD was incubated anaerobically (candle jar method) at 37°C for 24hrs to allow maintenance and growth of only NAD dependent organis. A loopful of enrichment culture was then streaked onto Blood agar (BA) with NAD, blood agar with feeder colony of S. aureus, Chocolate agar (CA) with NAD and CA with feeder colony (S. aureus) followed by anaerobic incubation for 24 hour and examined for the growth of characteristics colonies. Identification of bacteria - Colonial morphology of bacteria such as: shape, size, surface texture, edge, elevation and color was observed on pure culture; Gram staining and biochemical tests such as sugar fermentation, methyl red (MR), Voges-Proskauer (V-P), indole production, Catalase test and motility test in MIU medium were used foridentification of the bacteria. Antimicrobial susceptibility testing - The Bauer disk diffusion method was used to test antimicrobial susceptibility of Avibacterium paragallinarum using freshly prepared Mueller Hinton agar (Oxoid, UK). Antimicrobial agents and their disc concentrations used were as follows: ampicillin (10 μg), ciprofloxacin (5 μg), gentamicin (10 μg), trimethoprim-sulphamethoxazole (25 μg) and tetracycline (30 μg). Results of antibiotic sensitivity tests were recorded as sensitive, intermediate and resistant following the guidelines of Clinical and Laboratory Standard Institute.

Source of Information: Microbes and Health, June 2014, 3(1): 9-11; ISSN: 2226-0153 (Print) 2305-3542 (Online); http://journal.bsvmph.org/

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In Bangladesh infectious coryza is an important disease of economic significance occurring in multiage layer flocks but diagnosis of the disease still now based on postmortem examination of the dead birds.In the present study A. paragallinarumlike organism was isolation and identification of from Layer Chickens in Gazipur, Bangladesh. This organism grew more frequently with typical dewdrop like satellite colony when cultured on BA containing both NAD and feeder organism. Further in depth study on serogroup determination, molecular detection and characterization of A. paragallinarum is warranted.

Knowledge Management: Journal