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Isolation and Identification of Common Food Borne Bacteria from Spicy Puffed Rice and Determination of Their Antibiotic Resistance Profile

Md. Imran Rahman
Department of Microbiology, Hajee Mohammad Danesh Science and Technology University, Dinajpur5200, Bangladesh.

Md. Khaled Hossain
Department of Microbiology, Hajee Mohammad Danesh Science and Technology University, Dinajpur5200, Bangladesh.

Mst, Deloara Begum
Department of Microbiology, Hajee Mohammad Danesh Science and Technology University, Dinajpur5200, Bangladesh.

Nazmi Ara Rumi
Department of Microbiology, Hajee Mohammad Danesh Science and Technology University, Dinajpur5200, Bangladesh.

Md. Fakhruzzaman
Department of Microbiology, Hajee Mohammad Danesh Science and Technology University, Dinajpur5200, Bangladesh.

Md. Rafiqul Islam
Principal Scientific Officer, Livestock Division, Bangladesh Agricultural Research Council, Farmgate, Dhaka, Bangladesh.

Abstract/ Executive Summary:

The aim of this study was to determine bacterial load in street vended spicy puffed rice (jhalmuri), to isolate and identify the common food borne bacteria, and to know the antibiogram profile of the isolated bacteria. Samples were collected from different vendors at Sadar Upozila, Dinajpur, Bangladesh. Total viable bacteria were counted, and the samples were inoculated into various selective media. The bacteria were identified through cultural properties, staining characteristics and biochemical tests. Antibiotic sensitivity of the bacteria were conducted using disc diffusion method. Among 20 samples, all (100%) had bacterial contamination. The total viable count (TVC) in the samples were ranged from 8.4x10³ CFU/g to 3.24x10⁷ CFU/g. From the samples Pseudomonas aeruginosa (37.5%), Staphylococcus aureus (25%), E. coli (19.5%) Bacillus cereus (13%) and, Bacillus subtilis (5%) were isolated and identified. Antibiotic sensitivity test showed that Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa were sensitive to Chloramphenicol (60%) and Ciprofloxacin (60%). Bacillus cereus was moderately sensitive to Chloramphenicol, Ciprofloxacin and Kanamycin; E. coli was moderately sensitive to Chloramphenicol and Ciprofloxacin; Staphylococcus aureus and E. coli were found resistant to Kanamycin (40%). All isolates were found 100% resistant to Amoxycillin, Cefixime, Cephalexin, Neomycin and Ceftriaxone. Presence of coliforms in the sample might be due to poor quality of water, unhygienic vendor places and poor personal hygiene of vendors. The results of this study suggested that street vended ready to eat (RTE) foods should be manufactured under good hygienic practices.

Keywords: Antibiotic resistance, Bacterial load, Identification, Spicy puffed rice

Location: Bacteriology Laboratory of the Department of Microbiology, Faculty of Veterinary and Animal Science (VAS), Hajee Mohammad Danesh Science and Technology University (HSTU), Dinajpur, Bangladesh

Start Date: 00-01-2016

End Date: 00-07-2017

Program Area: Food Safety and Security

Commodity: Quality, Bacteria

Objectives:

The objectives of this research work were therefore to determine bacterial load in street vended spicy puffed rice, to isolate and identify the food borne bacteria from those samples and to know the antibiogram profile of isolated bacteria.

Methodology:

The present study was conducted during the period from January to July 2016, in the Bacteriology Laboratory of the Department of Microbiology, Faculty of Veterinary and Animal Science (VAS), Hajee Mohammad Danesh Science and Technology University (HSTU), Dinajpur. The samples were collected from different vendors located in Sadar Upozila, Dinajpur. A total number of 20 spicy puffed rice (jhalmuri) samples collected from different spicy puffed rice vendors. The bacteriological quality of samples was analyzed. The bacteriological analysis comprisee enumeration of TVC. Isolation of the samples was performed onto Mannitol salt (MS) agar, Eosin Methylene Blue (EMB) agar, MacConkey (MC) agar, Salmonella-Shigella (SS) agar. Identification of bacteria was performed by morphological, cultural and biochemical tests. The pattern of antibiotic sensitivity of bacterial isolates obtained from spicy puffed rice (jhalmuri) was studied. The materials used in the study comprise sample materials and laboratory materials. Sample materials include spicy puffed rice and associated instruments; those are clean rubber hand gloves, eppendorff tubes, and ice box containing ice. Different bacteriological culture media and reagents were for isolation and identification and also propagation of bacteria from spicy puffed rice. The culture media and reagents that were used in this experiment are Nutrient Broth (NB) was used as liquid culture media. Nutrient agar (NA), MacConkey (MC) agar medium. Eosin methylene blue (EMB) agar medium, Salmonella-Shigella (SS) agar medium and Manitol Salt (MS) agar medium were used as solid culture media. All of them were purchased from Hi Media, Mumbai, India. A total of 20 samples were collected for bacteriological examination. The street vendors on the busiest street areas were chosen for sample collection. The samples were collected from Basher hat, Gopalgonj, Ranigonj, HSTU High School, Dosmile. The samples were congregated during the months of January to July 2016. The time interval between sampling from each vendor was approximately 15 minutes. Two samples were collected on every sampling day. Food samples included cereal based (jhal-muri). Approximately 300 g of each food sample was collected using the vendors serving utensils, take parcel and placed into sterile plastic bags. All the collected samples were kept on an ice-box during transportation to the laboratory and stored at 4°C until testing. They were analyzed within 24 hours of sampling. The test organism was cultured in a medium containing tryptophan. The organisns break down tryptophan and indole is released. It was detected by the action of Kovac’s reagent or Ehrlich reagent (formation of red colored compound). This test was important for the identification of enterobacteria such as E. coli, P. rettgeri etc. Tryptophan containing broth was inoculated with bacteria. The tube was incubated at 37⁰ C for 24 hours. 0.5 ml of the Kovac’s reagent was added after the bacterial growth. In positive case after 2 minutes a red color ring appeared at the junction of medium and reagent in the tube. Individually isolated colonies of the same morphology were selected from appropriate agar plates, cloned and checked for purity of growth prior to characterization of the respective genera and species. Characteization into respective genera and species were done on the basis of morphological, cultural and biochemical reactions. The classification and specification of organisms was based on the scheme presented in Bergey’s Manual of Systematic Bacteriology (Holt, 1985). Stock culture was prepared and maintained for subsequent studies. Strict aseptic measures were maintained throughout the period of study.

Source of Information: Official journal of the Bangladesh Society for Safe Food, Volume 1 Issue 1, January 2018, Pages 33-47.

Article Link (Online Journal): http://bssf-bd.org/

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The recent study was conducted for the isolation and identification of food borne bacteria from spicy puffed rice study of their antibiotic sensitivity pattern. Among 20 spicy puffed rice (jhalmuri) samples, all (100%) had bacterial contamination. TVC in spicy puffed rice sample was ranged from 8.4x10³ CFU/g to 3.24x10⁷ CFU/g. From the spicy puffed rice Pseudomonas aeruginosa (37.5%), Staphylococcus aureus (25%), E. coli (19.5%) Bacillus cereus (13%) and, Bacillus subtilis (5%) were isolated and identified. Presence of coliforms in the sample might be due to poor quality of water, unhygienic vendor places and poor personal hygiene of vendors.. Antibiotic sensitivity test showed that Bacillus subtilis, Staphylococcus aureus and Pseudomonas aeruginosa were sensitive to Chloramphenicol (60%) and Ciprofloxacin (60%)but the Bacillus cereus and E. coli were moderately sensitive. Bacillus subtilis, Bacillus cereus were moderately sensitive to Kanamycin (40%) but the Staphylococcus aureus and E. coli were resistant. All isolates were found 100% resistant to Cefixime, Cephalexin, Neomycin, Amoxycillin and Ceftriaxone. The results of this study suggested that street vended RTE foods should be manufactured under good hygienic practices and conservation practices should be developed in order to minimize the microbial contamination of food. Moreover, it is recommended to develop an appropriate hazard analysis critical control point (HACCP) system, and a more close supervision of RTE food should be carried out by relevant authorities to enhance food safety.

Knowledge Management: Journal