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Isolation, Identification, and Antibiogram Studies of Salmonella Species and Escherichia Coli from Broiler Meat in Some Selected Areas of Bangladesh

Abu Saim Al-Salauddin
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Mohammad Farhad Hossain
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Amit Dutta
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Shahin Mahmud
Department of Biotechnology and Genetic Engineering, Faculty of Life Science, Mawlana Bhashani Science and Technology University, Santosh, Tangail-1902, Bangladesh.

Md. Shariful Islam
Department of Biotechnology and Genetic Engineering, Faculty of Life Science, Mawlana Bhashani Science and Technology University, Santosh, Tangail-1902, Bangladesh.

Sukumar Saha
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

S. M. Lutful Kabir
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Abstract/ Executive Summary:

A study was performed for the isolation, identification of Salmonella and Escherichia coli from broiler meat samples (leg muscle, breast muscle and drumstick) which were collected from different upazilla markets of Mymensingh, Gazipur, and Sherpur districts during the period of January 2015 to May 2015. A total of 60 samples were subjected to bacterial isolation and identification by using cultural, biochemical, and polymerase chain reaction assays. Using standard bacteriological techniques E. coli was isolated from 50 (83.33%) samples and Salmonella spp. from 18 (31.66%) samples. Furthermore, the isolates were subjected to antibiogram studies by disk diffusion method using eight commonly used antibiotics. Antibiogram studies revealed that gentamicin, ciprofloxacin, and norfloxacin were highly sensitive against all the isolated bacteria, whereas most of the isolates were resistant to amoxicillin, erythromycin, and tetracycline. Out of all the isolates, 5 isolates of E. coli and 3 isolates of Salmonella were found multidrug resistant. The study revealed the presence of multidrug resistant Salmonella and E. coli in broiler meat sold in live bird market of different upazillas studied; which deserves priority attention to prevent health damages of the consumers.

Keywords: Broiler meat, Salmonella spp., Escherichia coli, Antibiogram,Multidrug resistant, Antobiotic

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

Start Date: 00-01-2015

End Date: 00-05-2015

Program Area: Animal Health and Management

Commodity: Diseases

Objectives:

To isolate and identify the associated bacteria prevalent in broiler meat and
To find out the effective antibiotics against the bacteria through antibiogram studies.

Methodology:

A total of 60 dressed broiler carcasses were collected during the period of January 2015 to May 2015 and immediately brought to Bacteriology Laboratory of the Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh by maintaining cool chain using ice box. The samples were processed immediately for isolation and identification of Salmonella and E. coli. For the isolation of bacteria 10 g of meat (thigh muscle, breast muscle, and drumstick) were taken in a sterile pack and allowed to prepare meat homogenates by adding 90 ml of 0.1% peptone water using stomacher blender. The primary culture was performed in nutrient agar and nutrient broth media. For sub-culturing, suspected bacteria were inoculated separately onto different bacteriological media under the aseptic condition and incubated at 37°C for 24 hrs. Pure cultures were achieved by further sub culturing on selective agar. Cultural, morphological, and biochemical characteristics were studied to identify the bacterial flora. The cultural characteristics or colonial morphology of the bacteria grown on the Eosin Methylene Blue (EMB) and Xylose Lysine Deoxycholate (XLD) agar were recorded. Gram staining was performed to study the morphology and staining characteristics of the bacteria.Biochemical tests, such as sugar fermentation, methyl red (MR), voges-proskauer (VP), and indole tests, were performed to identify the bacteria tentatively. Bacterial DNA template was prepared by using boiling method. All the samples were examined by two pairs of primers to detect 16S rRNA gene of E. coli and histidine transport operon gene of Salmonella spp. Thermal profiles used in PCR are discussed. PCR products were separated on 2% agarose gel, stained with ethilium bromide and photographed using a Gel documentation system (BioRad). Antibiotic susceptibility test was performed by disk diffusion method using the commercial antibiotic disk on Mullar- Hinton agar to assess the susceptibility and resistance pattern of the isolates. For this purpose, eight different antibiotic discs were obtained from commercial sources (Himedia, India and Oxoid Ltd. England). The selected antibiotics used were ciprofloxacin (5 μg/disc), azithromycin (30 μg/disc), amoxicillin (30 μg/disc), gentamicin (10 μg/disc), norfloxacin (10 μg/disc), erythromycin (30 μg/disc), streptomycin (10 μg/disc), and tetracycline (30 μg/disc). The interpretation on susceptibility was done according to the guidelines of Clinical and Laboratory Standard Institute formerly known as NCCLS.

Source of Information: Int J Basic Clin Pharmacol. 4(5):999-1003, 2015 Oct; ISSN 2319-2003

Article Link (Online Journal): doi: http://dx.doi.org/10.18203/2319-2003.ijbcp20150881

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

E. coli and Salmonella spp. were isolated from the meat samples of broilers collected from different upazilla live bird market of Mymensingh, Gazipur, and Sherpur districts in Bangladesh. Prudent use of antibiotics should be considered in broiler production (only where permissible) since many strains are resistant to common antibiotics, and some were multidrug resistant as described in this study. Potential drug resistant pathogens in otherwise normal broilers might be a serious public health concern. Current findings warrants further studies with the isolated strains of bacteria.

Knowledge Management: Journal