Isolation of pathogenic Escherichia coli from buffalo meat sold in Parbhani city , Maharashtra , India

Aim: Isolation, characterization, in-vitro pathogenicity and antibiogram study of E.coli from buffalo meat sold in Parbhani city. Materials and Methods: Meat samples were collected from buffalo immediately after slaughter. Isolation, identification and enumeration of E. coli were done by following standard methods and protocols. Hemolysin test and Congo red binding assay were used to study in-vitro pathogenicity of E. coli isolates. Disc diffusion method was used to study antibiogram of pathogenic E. coli isolates. Results: A total of 250 buffalo meat samples were collected and processed. A total of 22 (8.80 percent) E. coli isolates were isolated with average differential count of 1.231 ± 0.136 log cfu/g on EMB agar. All the E. coli isolates were confirmed by 10 Grams staining, biochemical reactions and sugar fermentation and motility tests. A total of 9 (3.6 percent) E. coli isolates were found to be pathogenic by in-vitro pathogenicity testing. Antibiogram studies of pathogenic E. coli isolates showed that all 9 isolates were sensitive to gentamycin (20 ± 1.49 mm) while 7 isolate showed resistance to enrofloxacin (18.22 ± 3.58 mm) and tetracycline (11.44 ± 2.04 mm). Conclusion: Buffalo meat sold in Parbhani city is an important source of E. coli infection to human population. A total of 9 pathogenic E. coli were isolated from buffalo meat immediately after slaughter. All isolates were characterized and confirmed pathogenic by in-vitro pathogenicity tests. Antibiogram studies of all isolates revealed sensitivity to gentamicin and resistance to tetracycline and enrofloxacin.


Introduction
antimicrobial agents in maintenance of hygiene in animal husbandry practices as well as during India is a leading exporter of buffalo (Bubalus processing of meat.This results into development of bubalis) meat earning foreign exchange.The production drug resistant condition.Antibiotic resistance to beef of buffalo meat (beef) is increasing by 4.5 percent per borne E. coli is well documented [8,9].annum [1].According to Animal Husbandary statistical The growing problem of antibiotic resistant has database of Department of Animal Husbandary, Dairying become a significant public health concern world wide.and Fisheries (DADF), there are a total of 5,520 All uses of antimicrobial in human medicine and recognized and 4,707 unrecognized slaughter houses animal husbandry create selective pressure that favors in the country.The buffalo carcass has less fat, less emergence of antimicrobial resistance among microbone and higher proportion of muscle than cattle [2].organism [7].Sanitary indices refer to certain organisms or group of Keeping in view the public health importance and organisms which indicates presence of potentially hazards caused by pathogenic E. coli, the present study pathogenic or spoilage organisms in food.Coliform was undertaken to isolate and identify these pathogenic organisms in food indicate poor sanitary practices and and drug resistant organisms from buffalo meat sold in presence of other hazardous organisms.This group Parbhani city.includes E. coli and enumeration of E. coli counts is important to determine sanitary indices [2].Presence of

E. coli in food is an indicator of poor sanitary
Sample collection and processing: Buffalo beef samples conditions during processing.Isolation of E. coli from were collected from Parbhani Municipal Council meat and meat products is a common phenomenon [3, Abattoir.A total of 250 meat samples approximately 4].Studies showed that normal intestinal microflora weighing 50 gm were collected from neck site of specially commmensal E .colistrains under specific slaughtered animal in sterile polyethylene schachets condition, might serve as reservoir of resistance genes for the period from December 2011 to May 2012.that could be acquired by pathogenic bacteria [5].Since These meat samples collected were brought to the commensal bacterial strains are exposed to the same laboratory on ice within one hour and processed selective pressure as pathogenic strains, they might be immediately.used as an indicator of trends in antimicrobial resistance [6].Also E. coli being a member of Entero- st and identification of food borne Pathogens. 1 Edn.ICAR, Buffalo meat sold in Parbhani city is an important New Delhi, pp.55-69.13.Ishiguro E. E., T. Anisworth, J. T. Trust and W. W. Kay source of E. coli infection to human population.All (1985).Congo red, a differential medium for Aeromonas isolates were characterized and confirmed pathogenic salmonicida, detects the presence of the cell surface protein by in-vitro pathogenicity tests.Antibiogram studies of array involved in virulence J. of Bacteriol.164 (3): 1233.all isolates revealed sensitivity to gentamicin and 14.Anonymous (2001) Performance standards for antimicrobial susceptibility testing; National Committee of Clinical resistance to tetracycline and enrofloxacin.Laboratory Standards (NCCLS) Document M100-S11, Authors' contribution NCCLS, Pennsylvania,19087 USA.15.Hazarika R. A., D. K Singh , K. N. Kapoor, R. K. Agarwal The present work was carried out by CSS under the (2004a) Prevalence of different serotypes of Escherichia coli guidance of VV with the technical support of RW.RW in Buffalo meat and its product.Ind.J. Comparative Microbio., Immunology and Infec.Dises.25 (1) 19-22.drafted and revised the manuscript.All authors read 16.Clavero M., S. Rocelle; Beuchat, R. Larry and M. P. Doyle and approved the final manuscript.

( 1998 )
Thermal Inactivation of Escherichia coli O157:H7 Isolated from Ground Beef and Bovine Feces, and Suitability

In-vitro pathogenicity test of E. coli:
[4,26]antity of 10gm of beef positivity of hemolysin test and Congo red binding sample was minced with the help of sterile scissors and assay observed was 3.60 percent.All 9 pathogenic E. mixed in 90 ml normal saline solution (pH 7.2) in screw coli isolates were subjected to antibiotic sensitivity test -5 against gentamycin, tetracycline and enrofloxacin.The cap bottle.Ten fold serial dilutions were made up to 10 zones of inhibition of antibiotic were recorded.The dilution in normal saline solution (pH 7.2).A quantity In-vitro pathostudy are in agreement with earlier reports [3,15].genicitytest of confirmed E. coli isolates was done by The E. coli counts are in the of range 6.85 to 7.40 log 10 heamolysis and Congo red binding assay [12,13].cfu/g[16].Scanning of available literature reveals that Colonies showing hemolytic zone are considered as percentage of E. coli isolation and identification varies heamolytic positive.While isolates, producing intense considerably [17,18].The variation in percentage of E. Out of nine isolates 7 isolates shown fermentation and motility tests.All the 22 isolates were resistance to Enrofloxacin and Tetracycline.Similar subjected to in-vitro pathogenecity test that is reports in previous studies showed resistance of meat hemolysin test and Congo red binding assay.A total of borne E. coli to Gentamycin and Enrofloxacin [23], 9 E. coli isolates were found positive for pathogenecity Tetracycline and Ciprofloxacin [24,25].In present in both the in-vitro pathogenecity tests.The percent study sensitivity of E. coli to Tetracyclin was observed meat foods, drinking water and human diarrhoeic The resistance of pathogenic E. coli to the antibiotics samples of Panjab, India.Archiv.Clin.Microbiol.2:2.Bacteriological Analytical Manual, 8 edition animal husbandry practices.Pathogenic E. coli of meat publication by FDA, U.S. 11.Cowan S. T. and K. J. Steele (1993) Characters of Gram origin are always having grater potential to enter into positive bacteria.In Cowan and Steel's manual for Identififood chain.Buffalo meat, beef and other types of meat rd cation of Medical bacteria 3 edn.Cambridge Univ.Press.arecheapsource of human infection[4,26].
Isolation, identification and enumeration of E. coli: bacteriacae is being exposed to various antibiotics and Isolation of E. coli was done as per the method described in BAM [10Discussion i.e IMViC, catalase, oxidase & nitrate reduction and In present study, very low percentage( 08.80 sugar fermentation and motility tests.[11].percent) of E. coli was isolated.The findings of present from