Prevalence of Mycobacterium avium subspecies paratuberculosis, an animal pathogen, in the population of animal keepers of Ghaziabad and Saharanpur districts of North India using Multiple Diagnostic Tests
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Association of MAP in animal keepers with and without symptoms of inflammatory bowel disease (IBD)/Crohn's disease (CD) was studied. 131 clinical samples (25 stool, 53 blood and 53 serum) were collected from 54 Animal keepers having clinical profiles indistinguishable to IBD and were suspected for suffering with Crohn's disease. Animal keepers were residing in peri-urban areas of Ghaziabad (35) and Saharanpur (19) cities. Animal keepers had history of consumption of raw milk and were in contact with animals for variable periods (1–5, 6–10, 11–15 and >15 years) since birth. The 108 clinical samples (14 stool, 47 blood and 47 serum) were also collected from 47 Animal keepers (14 from Ghaziabad and 33 from Saharanpur) without the symptoms of IBD and were not suspected for suffering with CD. Of the 54 suspected CD animal keepers, 44, 41, 32, 23, 25 and 40 had history of abdominal pain, diarrhoeal episodes, weight loss, mild fever, smoking and consumption of raw milk, respectively. Stool samples were screened by microscopic examination and specific IS900 PCR. Blood samples were screened by PCR (blood PCR). Using ‘indigenous ELISA kit’, sero-prevalence of MAP was estimated in the two kind of animal keepers. DNA isolated from blood and stool samples were genotyped using IS1311 PCR-REA. Of the animal keeper suspected for CD, 36.0 (9/25), 28.0 (7/25), 12.9 (7/54), and 12.9% (7/54) were positive for MAP by microscopic examination, stool and blood PCR and ‘indigenous ELISA kit’, respectively. Of animal keepers not-suspected for CD, 14.2 (2/14), 7.1 (1/14), 2.1 (1/47) and 4.2% (2/47) were positive for MAP by microscopic examination, stool and blood PCR and ‘indigenous ELISA kit’, respectively. Of the 54 suspected animal keepers, 34.1% (15), 26.8% (11), 37.5% (12), 21.7% (5), 44.0% (11), and 40.0% (16) were positive for MAP and had history of abdominal pain, diarrhoeal episodes, weight loss, fever, smoking and consumption of raw milk, respectively. All the IS1311 PCR positive DNA samples were genotyped as ‘Indian Bison type’, which is major genotype infecting domestic livestock in the country. Presence of ‘Indian Bison type’ genotype as found in animals in India and higher prevalence of MAP infection in cases of suspected Animal keepers as compared to not-suspected Animal keepers, strongly indicated the association of Mycobacterium avium subspecies paratuberculosis in cases of Crohn's disesase in human beings in India.