Aim: The study was undertaken to isolate pathogenic E. coli from milk and various milk products, detection of virulence gene using Polymerase chain reaction (PCR) and investigate their antibiotic sensitivity pattern. Materials and Methods: Altogether 250 milk and various milk products samples consisting of raw milk (50), cheese (50), ice-cream (50), mawa (50) and dahi (50) were collected from milk vendors, retail shops located in Anand city, under aseptic precautions. For the enrichment of the organism from the collected samples, MacConkey broth was used and inoculation was carried out on MacConkey agar and EMB agar. Later on, to confirm the isolates, various biochemical tests such as IMViC test, Urease test were performed. Evaluation of antibiotic sensitivity pattern of E. coli was assessed by disk diffusion method. Finally the E. coli isolates were screened for the presence of virulence associated genes by PCR . Results: The prevalence of E. coli was observed 32 % in the samples comprising of milk (52.00%), cheese (28.00%), icecream (20.00%), mawa (44.00%), and dahi (16.00%). Antibiotic sensitivity was recorded high for Co-trimoxazole (100%) followed by Gentamicin (96.73%), Trimithoprime (93.47%) and Doxycycline hydochloride (92.39%). Least sensitivity was recorded for Ampicillin (8.69%). In this study, out of 80 E. coli isolates, 25 isolates (31.25%) were positive for stx genes, of which 7 (8.75%) isolates were positive for stx1 gene only, while 12 (15.00%) isolates were positive for stx2 gene only and 5 (6.25%) isolates were positive for both stx1 and stx2, 7 isolates (8.75%) were positive for eaeA gene and all the isolate were negetive for rfb O157 gene. Conclusions: Current study supports the finding that raw milk and various milk products can be regarded as critical source of pathogenic E. coli This explains the need of strict monitoring and surveillance for effective measures of hygiene and sanitary practice during production of milk and various milk products. [Vet World 2013; 6(8.000): 541-545
Aim: To detect the occurrence of pathogenic Escherichia coli from stool samples of diarrhoeal patients with history of raw milk consumption and to determine the public health significance of isolates, especially their role in causing human diseases. Materials and Methods:A total of 100 stool samples from diarrhoeal patients, with history of raw milk consumption were collected from primary health centres in and around Anand city, under aseptic conditions and a total of 50 raw milk samples were collected from milk vendors, retail shops located in Anand city in sterilized sample bottles. MacConkey broth was used for the enrichment of all the samples and inoculation was done on MacConkey agar and EMB agar was used as the selective media. This was followed by the confirmation of isolates using biochemical tests. For the serotyping, E. coli isolates were sent to the National Salmonella and Escherichia Centre, Central Research Institute (CRI), Kasauli, Himachal Pradesh. Detection of virulence genes was performed using PCR technique.Results: During the present investigation, 26 (52%) E. coli isolates from 50 milk samples and 59 (59%) E. coli isolates from 100 stool samples were recovered. Out of 85 E. coli isolates sent for serotyping, 74 isolates could be typed which were further distributed into 13 different serogroups O2, O4, O8, O17, O22, O25, O29, O36, O45, O60, O90, O116 and O172, whereas 8 isolates were found untypable and 3 isolates were reported rough isolates. Of the 59 E. coli isolates from stool samples of diarrhoeal patients tested, 15 isolates (25.42%) were reported to be positive for stx genes, among that 6 (10.16%) were positive for stx1 gene, 9 (15.25%) isolates were positive for stx2 gene, while 3 isolates (5.08%) were positive for eaeA gene. In this study, 21 E. coli isolates were found to be Shiga toxin producing E. coli (STEC) while none of the isolates were positive for the serotype O157. Conclusions:Our present findings indicate that raw milk may act as a source of pathogenic E. coli and it may be responsible for the occurrence of diarrhoea and various other health-related complications in humans. We therefore recommend proper managemental practices and effective control measures for improved hygiene and sanitation.
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