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Prevalence of Vibrio Cholerae in Human-, Poultry-, Animal Excreta and Compost Samples

Md. Aftab Uddin
Department of Microbiology, Stamford University Bangladesh, 51 Siddeswari Road, Dhaka 1217, Bangladesh

Rashed Noor
Department of Microbiology, Stamford University Bangladesh, 51 Siddeswari Road, Dhaka 1217, Bangladesh

Md. Wali Ullah
Department of Microbiology, Stamford University Bangladesh, 51 Siddeswari Road, Dhaka 1217, Bangladesh

Abstract/ Executive Summary:

Transmission of Vibrio cholerae might take place in water bodies that often undergo contamination with human-, poultry- and animal excreta. In the present study, wastes from all three categories were analyzed microbiologically. Twelve poultry excreta samples, 12 human excreta samples and 12 cowdung samples were examined for the presence of V. cholerae. High prevalence of V. cholerae was found in each of the samples and the highest prevalence was observed for human excreta samples (2.2x10⁸ cfu/g). Besides, 24 compost samples, which were prepared using human feces, were tested. The load of V. cholerae was found lower in the compost samples (from 0 to 2.2x10³ cfu/g) than that of the excreta samples. The study of antibiogram showed that all the isolates were resistant against ampicillin, erythromycin, tetracycline, chloramphenicol, nalidixic acid, meropenem and cefotaxime.

Keywords: Vibrio cholerae; Human excreta; Personal hygiene; Domestic animal; Compost

Location: Department of Microbiology, Stamford University Bangladesh

Start Date:

End Date:

Program Area: Pest Management

Commodity: Poultry

Objectives:

1. The present study was designed to isolate, identify and quantify V. cholerae from human excreta, poultry excreta and cow dung. The results were compared with that of compost samples. The antibiotic susceptibility patterns of the isolates were also investigated.

Methodology:

Excreta sample collection. Twelve human excreta and 12 poultry excreta samples were collected using sterile cotton swab. Cow dung samples were collected with the help of sterile spatula. Preparation and collection of Compost Samples. Pits having size of 3' in diameter and 4' in length were constructed for composting. Human excreta were dumped inside the pits. Trace amount of ash or lime was mixed with the excreta. The pits were covered with leaf and grass which were then allowed to stay for at least 6 months for composting. A total of 24 compost samples were then collected in sterile plastic container and transported immediately to the laboratory. Enrichment and Isolation of V. cholerae. Samples were enriched in alkaline peptone water (APW) (Difco Laboratories, Detroit, Mich.) at 37 °C for 6 to 8 h before isolation. Enriched samples were then subjected to 10 fold serial dilution up to 10-5. From 10-2 and 10-4 dilutions, 0.1 ml samples were inoculated onto thiosulfate citrate bile salt sucrose (TCBS) agar (Difco Laboratories, Detroit, Mich) and incubated at 37⁰C for 18 to 24 h. Moreover, nutrient agar was also used for the determination of total viable bacteria. Confirmation of V. cholerae. After incubation, sucrose fermenting yellow colored colonies on TCBS agar were further streaked on to taurocholate tellurite gelatin agar (TTGA) for confirmation of V. cholerae and incubated at 37°C for 48 hours. Characteristic black centered colonies were selected and screened for the presence of cytochrome oxidase and gelatinase activity by subculturing onto gelatin agar media. All the colonies, which were found to harbor cytochrome oxidase and showed gelatinase activity, were then taken for further confirmation by biochemical tests (triple sugar iron (TSI) agar test, motility test, Voges-Proskauer (VP) test and salt tolerance test) according to ‘Microbiological Laboratory Manual’. Determination of antimicrobial susceptibility by modified Kirby-Bauer method. All the isolates of V. cholerae were tested for antibiotic susceptibility against 10 antibacterial drugs by modified Kirby-Bauer method (19) on Mueller-Hinton Agar (Difco, Detroit, MI) media using antibiotic discs (Neo- Sensitabs, Rosco, Denmark) including chloramphenicol (30 μg), nalidixic acid (30 μg), meropenem (30 μg), cefotaxime (30 μg), ampicillin (10 μg), erythromycin (100 μg), neomycin (120 μg), norfloxacin (10 μg), imipenem (10 μg) and tetracycline (10 μg). The zone diameters for individual antimicrobial agents were then translated into susceptible, intermediate or resistant categories according to the interpretation table supplied by the manufacturer (Oxoid limited, England).

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 study envisaged on the urge of composting human-, animal- and poultry excreta which may further ensure a hygienic waste disposal system also being of agro-economic importance. The elimination of V. cholerae in the compost samples compared to the excreta samples refer to a safe and natural destruction of microorganisms in the environment, thus reducing the possibility of transmission of V. cholerae.

Knowledge Management: Journal