Lacto-prevalence, genotyping of Mycobacterium avium subspecies paratuberculosis and evaluation of three diagnostic tests in milk of naturally infected goatherds

et al. 2010

Trop Anim Health Prod

Genotyping of Mycobacterium avium subspecies paratuberculosis (MAP) is important for precise classification of bacterium and for understanding the molecular epidemiology. The present study reports detection and typing of the MAP from milk. On the basis of clinical signs of diarrhea and/or weakness, the dairy animals suspected for Johne's disease were screened by Ziehl-Neelsen staining of fecal samples. The milk samples from 13 selected animals were processed for DNA extraction and direct IS900 polymerase chain reaction (PCR). MAP identified by IS900 PCR was genotyped using IS1311 PCR-restriction endonuclease analysis (REA). IS900 milk PCR revealed 30.8% animals positive for MAP, including 40% of the moderate and 50% of the heavy fecal shedders. All infected animals showed Bison type MAP in IS1311 PCR-REA. IS900 PCR can be used for screening of milk for MAP; however, the method needs to be evaluated for subclinical cases. IS1311 PCR-REA results indicated the predominance of Bison type MAP in the dairy animals of this region.

Section: Resultsmentioning

confidence: 99%

Molecular detection and typing of Mycobacterium avium subspecies paratuberculosis from milk samples of dairy animals

Kaur

Filia

et al. 2010

Trop Anim Health Prod

“…Combinations of serum ELISA and milk ELISA, faecal microscopy and faecal PCR, milk ELISA and milk PCR, and faecal PCR and serum ELISA were applied on 204, 68, 207, and 62 dairy cows, respectively, to evaluate best combination for the diagnosis of Johne's disease in dairy cattle herds. Whey was curdled with 3.0% citric acid and centrifuged at 5000 rpm for 30 minutes to collect clear whey, which was stored at −20°C till further use for the presence of MAP antibodies by “Indigenous ELISA Kit” [ 9 ].…”

Section: Methodsmentioning

confidence: 99%

Comparative Evaluation of Different Test Combinations for Diagnosis ofMycobacterium aviumSubspeciesparatuberculosisInfecting Dairy Herds in India

Garg

Patil

BioMed Research International

et al. 2015

A total of 355 cows were sampled (serum, n = 315; faeces, n = 355; milk, n = 209) from dairy farms located in the Punjab state of India. Faeces and serum/milk samples were screened by acid fast staining and “indigenous ELISA,” respectively. IS900 PCR was used to screen faeces and milk samples. Bio-load of MAP in dairy cows was 36.9, 15.6, 16.3, and 14.4%, using microscopy, serum ELISA, milk ELISA and milk PCR, respectively. Estimated kappa values between different test combinations: serum and milk ELISA, faecal microscopy and faecal PCR, milk ELISA and milk PCR, faecal PCR and serum ELISA were 0.325, 0.241, 0.682, and 0.677, respectively. Estimation of the relative sensitivity and specificity of different tests in the present study indicated that “serum ELISA” and “milk ELISA” were good screening tests, add “milk PCR” was “confirmatory test” for MAP infection. Combination of milk ELISA with milk PCR may be adopted as a model strategy for screening and diagnosis of JD in lactating/dairy cattle herds in Indian conditions.

“…Goats Raw milk: Screening of the raw milk samples of goats belonging to CIRG farm herds (with herds endemic for JD since 1985 (Singh et al 2014a)), the percent bio-load of MAP was 20.0-69.8, 7.6-37.7, 30.7-54.7 and 13.8 using milk culture, milk IS900 PCR, milk ELISA and milk microscopy (Singh and Vihan 2004;Kumar et al 2008;Raghuvanshi et al 2010).…”

Section: Map In Dairy Productsmentioning

confidence: 99%

Mycobacterium avium subspecies paratuberculosis – an important food borne pathogen of high public health significance with special reference to India: an update

Chaubey

Gupta

et al. 2017

Veterinary Quarterly

This review underlines the public health significance of 'Indian Bison Type' of Mycobacterium avium subspecies paratuberculosis (MAP) and also its potential as 'zoonotic infection'. In the absence of control programs, bio-load of MAP is increasing and if we take total population of animals (500 million plus) and human beings (1.23 billion plus) into account, the number of infected animals and human beings will run into millions in India. Our research on screening of over 26,000 domestic livestock for MAP infection using 4 different diagnostic tests (microscopy, culture, ELISA and PCR), during last 31 years has shown that the average bio-load of MAP in the livestock population of India is very high (cattle 43%, buffaloes 36%, goats 23% and sheep 41%). 'Mass screening' of 28,291 human samples between 2008-2016 revealed also high bio-load of MAP. It has been proved that MAP is not in-activated during pasteurization and therefore live bacilli are continuously reaching human population by consumption of even pasteurized milk and other milk products. Live bacilli have also been recovered from meat products and the environment thus illustrating the potential of MAP as pathogen of public health concern. However, at present, there is inadequate scientific evidence to confirm a conclusive link between MAP infection and Johne's disease in ruminants and some cases of Crohn's disease in human beings.