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Detection of Multidrug Resistance Aeromonas Hydrophila in Farm Raised Fresh Water Prawns

Md. Bakhtiar Lijon
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Mst. Mousumi Khatun
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Ariful Islam
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Mst. Minara Khatun
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Md. Ariful Islam
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

This study was undertaken for isolation, identification and determination of antibiogram profile of Aeromonas hydrophila in farm raised fresh water prawn (Macrobrachium rosenbergii) on five commercial ghers. Fresh water prawns (n=25) were collected from five ghers located at Satkhira, Bagerhat and Khulna districts of Bangladesh. Brain (n=25), muscle (n=25) and intestine (n=25) samples were collected aseptically from fresh water prawn and inoculated into alkaline peptone (APW) water for enrichment at 37⁰ for 8 h. Enriched cultured was streaked into Thiosulfate Citrate Bile Salt Sucrose (TCBS) agar to isolate bacteria. Bacteria were identified by cultural, Gram staining, biochemical properties and polymerase chain reaction (PCR) assay. The antibiogram profiles of bacteria were investigated against 5 commonly used antibiotics (Gentamicin, Cefalexin, Ampicillin, Azithromycin and Ciprofloxacin) by disc diffusion method. Thirteen A. hydrophila isolates were identified and the prevalence of the A. hydrophila in fresh water prawn was 17.33%. All 13 (100%) isolates were sensitive to Ciprofloxacin, Gentamicin, Azithromycin and resistant to Ampicillin and Cefalexin. The results of this study indicate that farm raised fresh water prawn harbor multidrug resistant A. hydrophila which might causes public health problem if enter into human food chain.

Keywords: Aeromonas hydrophila, Antibiogram profile, Fresh water prawn, Prevalence, PCR assay

Location: Satkhira, Bagerhat and Khulna districts, Bangladesh.

Start Date: 00-01-2014

End Date: 00-06-2014

Program Area: Animal Health and Management

Commodity: Prawn

Objectives:

The objectives of this study were

Isolation and identification of A. hydrophila from farm raised fresh water prawn, and
determination of antibiogram profile of A. hydrophila against five commonly used antibiotics.

Methodology:

Collection of samples: A total of 25 fresh water prawns were collected from five ghers which were located at Satkhira (n=2, Satkhira Sadar and Assasuni Upazillas), Bagerhat (n=2, Bagerhat Sadar and Rampal Upazillas) and Khulna (n=1, Dumuria Upazilla) districts of Bangladesh during the period from January to June, 2014. The samples were packed into sterile polyethene bags in an ice box and transported to the Department of Microbiology and Hygiene at the Bangladesh Agricultural University (BAU), Mymensingh for bacteriological study. Processing of samples: Brain (n=25), muscle (n=25) and intestinal (n=25) specimens of prawns were aseptically collected. Muscle samples (0.5 g) were mixed with 4.5 mL alkaline peptone water (APW) and grinded by pastle and morter to prepare a 10% tissue suspension. Intestinal samples were cut by scissors into several small pieces and homogenized in PBS to prepare a 10% tissue suspension. Enrichment of samples: A loopful of brain fluid was inoculated into 4.5 mL of APW and incubated at 37 for 8 h. Suspension of muscle (0.5 mL) and intestine (0.5 mL) were separately inoculated into test tubes containing 4.5 mL APW and incubated at 37 for 8 h. Isolation of bacteria: One loopful of enrichment culture of brain, muscle and intestine was separately streaked duplicate onto Thiosulfate citrate bile salts sucrose (TCBS) agar (Himedia, India) and incubated aerobically at 37 for 24 h. Single colony grown onto the TCBS agar was further sub cultured onto TCBS agar until pure cultures were obtained. Identification of bacteria: Identification of bacteria was conducted by observing cultural characteristics and colony morphology on the TCBS agar and growth of bacteria into nutrient broth containing 0% and 6% sodium chloride (NaCl). Gram’s staining method, motility test, sugar fermentation and biochemical tests (oxidase test, catalase test, citrate test, indole test and MR-VP test) were performed to identify bacteria. Molecular detection of bacteria by PCR: A genus specific PCR assay was performed to identify Aeromonas spp. by amplifying 276-bp fragment of lipase gene (Delamare et al., 2012). Antibiotic sensitivity test: Antibiogram profile of 13 A. hydrophila isolates was done against five different antibiotic such as: Gentamicin, Azithromycin, Ciprofloxacin, Ampicillin and cefalexin (Himedia, India). The antibiotics sensitivity testing was carried out according to instructions of the Clinical and Laboratory Standard Institute (CLSI, 2011).

Source of Information: J. Adv. Vet. Anim. Res., 2(4): 469-474, Volume 2 Issue 4 (December 2015)

Article Link (Online Journal): http://www.banglajol.info/index.php/PTCB/article/download/26252/17633

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Data of this study suggest that A. hydrophila are prevalent in fresh water farm raised prawn in the ghers of the southern part of the Bangladesh. Data of antibiogram study suggest that farm raised fresh water prawn harbors multidrug resistant A. hydrophila which may cause public health hazard if enter into the food chain.

Knowledge Management: Journal