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Frequency of Drug Resistant Salmonella Spp. Isolated from Poultry Samples in Bangladesh

Muhammad Ali Akond
Department of Botany, Jahangirnagar University, Dhaka-1342, Bangladesh

Saidul Alam
Department of Botany, Jahangirnagar University, Dhaka-1342, Bangladesh

S. M. R. Hassan
Department of Botany, Jahangirnagar University, Dhaka-1342, Bangladesh

Md. Mahbubur Rahman
Laboratory Sciences Division, icddr,b, Dhaka-1212, Bangladesh

Mozammel Hoq
Department of Microbiology, University of Dhaka,Dhaka-1000, Bangladesh

Momena Shirin
Institute of Public Health, Mohakhali, Dhaka-1212,Bangladesh

Abstract/ Executive Summary:

Present study was conducted to determine the levels of Salmonella spp. mediated contamination in samples collected from poultry and poultry environments in Bangladesh and to determine the antibiotic resistance trait in those pathogens. A total of 300 samples were collected from five different sources (cloacal swab, intestinal fluid, egg surface, hand wash of chicken handler and soil of chicken market) and 80 (26.6%) samples were found to be contaminated with Salmonella spp. One hundred and fifty Salmonella strains were isolated among which, 91 were detected as S. enteriditis and 59 as S. typhimurium through morphological, biochemical and serological tests. Susceptibility of the isolates against 13 antimicrobials was tested, and resistance against chloramphenicol (30 μg), erythromycin (15 μg), ampicillin (10 μg), riphampicin (5 μg), cephalexin (15 μg), cefixine (5 μg), penicillin (10 μg), tetracycline (30 μg), norfloxacin (10 μg), nalidixic acid (30 μg) and ciprofloxacin (10 μg) was found in 58%, 82%, 88%, 60%, 65%, 50%, 100%, 100%, 20%, 20% and 20% of the strains, consecutively. 76% isolates were found to be sensitive against gentamicin and 70% against streptomycin. Some of the isolates interestingly exhibited multiple-drug resistances against at least 6 to 10 antibiotics. The results indicated the higher resistance of Salmonella spp. against antibiotics which serving as a threat not only to poultry industry of Bangladesh but also possesses a serious threat to public and animal health.

Keywords: Salmonella spp.; drug resistance; poultry environments

Location: Department of Botany, Jahangirnagar University, Dhaka-1342, Bangladesh

Start Date: 00-10-2008

End Date: 00-06-2010

Program Area: Food Safety and Security

Commodity: Chicken

Objectives:

1. The study was focused on the determination of the level of Salmonella contamination in the poultry environments and their resistance against different antibiotics in Bangladesh.

Methodology:

Sampling Area and Sample Collection. The study was carried out during the period from October, 2008 to June, 2010. Three hundred samples from cloacal swabs of chicken (n = 80), intestinal fluid of chicken (n=70), egg surface (n = 60), soil of chicken market (n=40) and hand wash of chicken handlers (n=50) were collected from various poultry shops of 4 City Corporation markets viz., Mohakhali, Dhaka New Market, Mirpur and Malibag of the Dhaka metropolitan city. The samples were collected following the procedure. Transportation of Sample. All the samples were transported to the laboratory immediately after collection in an insulating foam box with ice maintaining the temperature ranging from 4⁰C to 6⁰C. In case of the cloacal swab samples, the test tubes containing selenite broth were incubated for 6 hours at 37⁰C immediately after coming to the laboratory for the enrichment of the Salmonella spp. Isolation of Salmonella spp. Serial dilutions up to 10–3 were made for soil sample. 0.1 ml of sample from intestinal fluid, 0.2 ml of sample prepared from soil of chicken market (dilution up to 10–3) and a loopfull of selective selenite broth from previously incubated cloacal swab sample were spread on the solid surface of Salmonella-Shigella (SS) agar medium (Hi-Media, India). A total of 1.0 ml sample from intestinal fluid and soil of chicken market was placed onto sterile plates which was then mixed with sterile medium poured into the plates after being cooled to about 42-45⁰C. About 10-100 ml of water sample from egg surface and hand wash of chicken handlers was filtrated through the membrane filter (0.45 μm, Millipore) to isolate the organism. The membrane filter was then placed on the surface of Salmonella-Shigella (SS) agar medium. Replications of all samples using triplicate plates were tested for successful isolation of typical colonies by incubating them at 37⁰C for 24 hours. Biochemical Identification and Serotyping of Salmonella spp. A total of 150 strains were subjected for biochemical identification including Gram staining, urease test, catalase test, oxidase test, H₂S production test, lactose utilization test, indole test, Voges-Proskauer test and citrate utilization test. Moreover, Salmonella serotyping was determined by latex agglutination tests using polyvalent antisera (Denka Seiken Co. Ltd, Tokyo, Japan), according to the manufacturer’s instruction. The presumptive Salmonella isolates were mixed with the latex test reagent and was observed for the agglutination reaction to check the presence of Salmonella. Antimicrobial Susceptibility Test. A total of 100 selected Salmonella strains, recovered from cloacal swab, intestinal fluid, handwash of chicken handler, egg surface and soil sample from chicken market, were tested for resistance against antimicrobial agents by Kirby-Bauer method using Mueller-Hinton agar plates. The standard procedure of the Clinical and Laboratory Standards Institute (CLSI) were strictly followed throughout the testing procedure. Quality control strain Escherichia coli ATCC 25922 was included in each run. All Salmonella isolates were subjected against 14 commercial antibiotic discs (Becton Dickinson Antibiotic Disc, U.S.A.) of streptomycin (10 μg), erythromycin (15 μg), chloramphenicol (30 μg), ciprofloxacin (10 μg), tetracycline (30 μg), penicillin (10 μg), norfloxacin (10 μg), cephalexin (15 μg), riphampicin (5 μg), cefixine (5 μg), ampicillin (10 μg), nalidixic acid (30 μg) and gentamicin (10 μg). The isolates were categorized as susceptible, intermediate and resistant according to the zone diameter interpretative standards recommendations by CLSI.

Source of Information: Stamford Journal of Microbiology, December 2012. Vol. 2, Issue 1; ISSN: 2074-5346

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The results of the study have illustrated the extent of antibiotic resistance in Salmonella serotypes from poultry sources in Bangladesh. The domestic and commercial handlers of poultry and poultry products in chicken shops and household and the peoples engaged in the poultry farms need more attention to strictly follow the rules and guidelines of hygiene to reduce or eliminate the risk of antibiotic resistant Salmonella and other pathogenic microbes. In addition, the use of antibiotics both for farming and for medication should be astute to minimize the chance for organisms to develop resistance.

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