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Research Detail

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Md. Atiqur Rahman Sarker
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Md. Mazedul Haque
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Rafia Afroze Rifa
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Fateha Akther Ema
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

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

Aim: The study was conducted for the isolation, identification, and antibiogram of bacteria obtained from fresh guava (Psidium guajava). Materials and Methods: A total of 25 fresh guavas were collected from five markets located in Mymensingh city. Guava samples were cultured onto various selective media such as eosin methylene blue, xylose lysine deoxycholate, thiosulfate citrate-bile salts-sucrose, blood agar, and mannitol salt agar for the isolation of bacteria. Biochemical tests (dextrose, maltose, lactose, sucrose, mannitol, methyl red, Voges–Proskauer, and indole) were performed to identify the bacteria. Results: Total viable counts of guava were ranged between log 6.56 colony-forming unit (cfu)/ml and 6.62 cfu/ml. A total of 106 bacterial isolates belonged to five genera (Escherichia coli, Salmonella spp., Vibrio spp., Bacillus spp., and Staphylococcus spp.) were identified. Salmonella spp. (23.6%) was the most prevalent, followed by E. coli (22.64%), Bacillus spp. (19.81%), Staphylococcus spp. (17.92%), and Vibrio spp. (16.03%). The results of antibiotic sensitivity test showed that Salmonella spp., Bacillus spp., and E. coli were sensitive to chloramphenicol, ciprofloxacin, and gentamicin and resistant to ampicillin and cephalexin. Vibrio spp. was sensitive to chloramphenicol and gentamicin, intermediately sensitive to ciprofloxacin and ampicillin and resistant to cephalexin. Conclusion: The results of this study indicate that fresh guava contains multidrug-resistant bacteria which might pose a public health risk.

  Antibiogram profile, Bacteria, Guava, Identification, Isolation, Mymensingh.
  In the Bacteriology Laboratory of the Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University (BAU), Mymensingh.
  00-07-2013
  00-12-2013
  Pest Management
  Guava

It is evident that no survey or assessment of guava fruit safety has yet been conducted in Bangladesh. Therefore, the objective of the present study was to isolate, identify and antimicrobial profile of bacteria isolated from fresh guava fruits sold at five markets in Mymensingh city.

Ethical approval The present study was conducted during the period from July to December 2013, in the Bacteriology Laboratory of the Department of Microbiology and  Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University (BAU), Mymensingh.

Collection and transportation of samples Guava samples were collected from KR market, Sheshmor, Kewatkhali, Meshubazar, and Nutunbazar. Samples were collected in sterile polythene bags separately. The samples were transported carefully to the Bacteriology Laboratory of the Department of Microbiology and Hygiene, BAU, Mymensingh, for bacteriological analysis.

Processing of guava samples Each guava sample was washed with 20 ml sterile PBS and transferred the washing sample into a separate poly bag. A ten-fold serial dilution of the samples was performed in the nutrient broth.

Determination of total viable count (TVC) 0.5 ml of each 10-fold diluted sample was transferred and spread on Plate Count Agar using a sterile pipette and a sterile glass spreader. Then, the plates were kept in an incubator at 37°C for 24-48 h. The number of colonies in a particular dilution was multiplied by the dilution factor to obtain TVC which was expressed as a mean log10 ± standard deviation colony-forming unit (cfu) per ml.

Isolation of bacteria in pure culture The isolation and identification of bacteria were performed according to the method described by Carter [6]. Samples were enriched in nutrient broth at 37°C for 24 h. The overnight cultures were streaked on SS agar (for Salmonella spp.), eosin methylene blue (EMB) (for Escherichia coli), BA (for Bacillus spp.), MS agar (for Staphylococcus spp.), and thiosulfate-citrate-bile salts-sucrose (TCBS) agar (for Vibrio spp.). The inoculated plates were incubated at 37°C for 24 h. A single colony was further subcultured until pure culture was obtained. Identification of bacteria was performed on the basis of colony morphology, Gram’s staining reaction, motility test, and biochemical tests.

Molecular detection of E. coli by polymerase chain reaction (PCR) DNA extraction A pure bacterial colony of E. coli was mixed with 100 µl of distilled water which was boiled for 10 min and then immediately kept on ice for cold shock. Finally, centrifugation was done at 10,000 rpm for 10 min. The supernatant was collected and used as a DNA template for PCR.

Primers used for PCR A genus-specific PCR was performed to amplify 16S rRNA of E. coli using previously published primers.

Antibiotic sensitivity test Five isolates randomly selected from five genera were tested for antimicrobial drug susceptibility against five commonly used antibiotics by disc diffusion or Kirby-Bauer method [8]. Selection of 3–5  well-isolated colonies from the SS, BA, EMB, MS, and TCBS agar plate. Touched the top of colony with a loop and the growth is transferred into the nutrient broth. The broths were streaked onto Mueller-Hinton agar plates using sterile glass spreader homogeneously. Then, the antibiotic disc was placed onto Muller-Hinton agar and incubated at 37°C for 24 h. The plates were examined, and the diameter of the zones of inhibition was measured in mm from the edge of the disc to the edge of the zone using a meter ruler.

Statistical analysis The results of TVC of the bacteria of guava sold at local markets were analyzed for statistical significance using [9] multiple range test (SPSS 11.5, US). A p≤0.01 was considered to be statistically significant. 

  Veterinary World, Vol.11/August-2018 EISSN: 2231-0916
  Available at www.veterinaryworld.org/Vol.11/August-2018/19.pd
Funding Source:
1.   Budget:  
  

Data of this study suggest that guava sold at local markets of Mymensingh city harbors multidrug-resistant bacteria which underscore the need of the implementation of proper hygienic measures during pre- and post-harvest stages of production and at the time of transportation, storage, selling, and preparation of different products such as juices to safeguard public health.

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