Kundan Kumar Chaubey

Peste Des Petits Ruminants Virus Infection of Small Ruminants: A Comprehensive Review

Peste des petits ruminants (PPR) is caused by a Morbillivirus that belongs to the family Paramyxoviridae. PPR is an acute, highly contagious and fatal disease primarily affecting goats and sheep, whereas cattle undergo sub-clinical infection. With morbidity and mortality rates that can be as high as 90%, PPR is classified as an OIE (Office International des Epizooties)-listed disease. Considering the importance of sheep and goats in the livelihood of the poor and marginal farmers in Africa and South Asia, PPR is an important concern for food security and poverty alleviation. PPR virus (PPRV) and rinderpest virus (RPV) are closely related Morbilliviruses. Rinderpest has been globally eradicated by mass vaccination. Though a live attenuated vaccine is available against PPR for immunoprophylaxis, due to its instability in subtropical climate (thermo-sensitivity), unavailability of required doses and insufficient coverage (herd immunity), the disease control program has not been a great success. Further, emerging evidence of poor cross neutralization between vaccine strain and PPRV strains currently circulating in the field has raised concerns about the protective efficacy of the existing PPR vaccines. This review summarizes the recent advancement in PPRV replication, its pathogenesis, immune response to vaccine and disease control. Attempts have also been made to highlight the current trends in understanding the host susceptibility and resistance to PPR.

Trends and advances in the diagnosis and control of paratuberculosis in domestic livestock.

ABSTRACT Paratuberculosis (pTB) is a chronic granulomatous enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP) in a wide variety of domestic and wild animals. Control of pTB is difficult due to the lack of sensitive, efficacious and cost-effective diagnostics and marker vaccines. Microscopy, culture, and PCR have been used for the screening of MAP infection in animals for quite a long time. Besides, giving variable sensitivity and specificity, these tests have not been considered ideal for large-scale screening of domestic livestock. Serological tests like ELISA easily detects anti-MAP antibodies. However, it cannot differentiate between the vaccinated and infected animals. Nanotechnology-based diagnostic tests are underway to improve the sensitivity and specificity. Newer generation diagnostic tests based on recombinant MAP secretory proteins would open new paradigm for the differentiation between infected and vaccinated animals and for early detection of the infection. Due to higher seroreactivity of secretory proteins vis-à-vis cellular proteins, the secretory proteins may be used as marker vaccine, which may aid in the control of pTB infection in animals. Secretory proteins can be potentially used to develop future diagnostics, surveillance and monitoring of the disease progression in animals and the marker vaccine for the control and eradication of pTB.

Genome Sequence of the “Indian Bison Type” Biotype of Mycobacterium avium subsp. paratuberculosis Strain S5

ABSTRACT We report the 4.79-Mb genome sequence of the “Indian Bison Type” biotype of Mycobacterium avium subsp. paratuberculosis strain S5, isolated from a terminally sick Jamunapari goat at the CIRG (Central Institute for Research on Goats) farm in India. This draft genome will help in studying novelties of this biotype, which is widely distributed in animals and human beings in India.

Concurrent Resolution of Chronic Diarrhea Likely Due to Crohn's Disease and Infection with Mycobacterium avium paratuberculosis.

Examination of samples of stool from a 61-year-old male patient, presenting with the clinical symptoms of Crohn’s disease (CD), revealed massive shedding of acid fast bacilli with the morphology of Mycobacterium avium paratuberculosis (MAP), the causative agent of Johne’s disease in cattle. MAP was cultured from the stool. Biotyping of the bacterium isolated from cultures of stool demonstrated, it was the Indian Bison biotype of MAP, the dominant biotype infecting livestock and humans in India. Based on this finding and because the patient was unresponsive to standard therapy used in India to treat patients with gastrointestinal inflammatory disorders, the patient was placed on a regimen of multi-antibiotic therapy, currently used to treat tuberculosis and CD. After 1 year of treatment, the patient’s health was restored, concurrent with cessation of shedding of MAP in his stool. This patient is the first case shown to shed MAP from the stool who was cured of infection with antibiotics and who was concurrently cured of clinical signs of CD.

Comparative Evaluation of Different Test Combinations for Diagnosis of Mycobacterium avium Subspecies paratuberculosis Infecting Dairy Herds in India

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.

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

ABSTRACT 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 Johnes disease in ruminants and some cases of Crohns disease in human beings.

The Consensus from the Mycobacterium avium ssp. paratuberculosis (MAP) Conference 2017

On March 24 and 25, 2017 researchers and clinicians from around the world met at Temple University in Philadelphia to discuss the current knowledge of Mycobacterium avium ssp. paratuberculosis (MAP) and its relationship to human disease. The conference was held because of shared concern that MAP is a zoonotic bacterium that poses a threat not only to animal health but also human health. In order to further study this problem, the conferees discussed ways to improve MAP diagnostic tests and discussed potential future anti-MAP clinical trials. The conference proceedings may be viewed on the www.Humanpara.org website. A summary of the salient work in this field is followed by recommendations from a majority of the conferees.

Inhibitor of sarco/endoplasmic reticulum calcium-ATPase impairs paramyxovirus replication

Sarco/endoplasmic reticulum calcium-ATPase (SERCA) is a membrane bound cytosolic enzyme that is known to regulate the uptake of calcium into the sarco/endoplasmic reticulum. Herein, we demonstrate for the first time that SERCA can also regulate paramyxovirus [Peste des petits ruminants virus (PPRV) and Newcastle disease virus (NDV)] replication. Treatment of Vero cells with SERCA specific inhibitor (Thapsigargin) at a concentration that is nontoxic to the cells significantly reduced virus replication. Conversely, overexpression of SERCA rescued the inhibitory effect of Thapsigargin on virus replication. PPRV/NDV infection induced SERCA expression in Vero cells which could be blocked by Thapsigargin. With the help of time-of-addition and virus step-specific assays, it was observed that Thapsigargin specifically inhibits viral entry and subcellular localization of the viral proteins. Furthermore, NDV, but not PPRV acquired a significant resistance to Thapsigargin on long-term passage (P=70) in Vero cells. To the best of our knowledge, this is the first report describing virus supportive role of SERCA and a rare report suggesting that viruses may acquire resistance even in the presence of an inhibitor that targets a cellular factor. This study will contribute in understanding paramyxovirus replication and development of antiviral therapeutics using SERCA (host factor) as a candidate drug target.

BIO-INCIDENCE OF Mycobacterium avium SUBSPECIES paratuberculosis IN THE PASTEURIZED LIQUID MILK, FLAVOURED MILK AND MILK POWDER COMMERCIALLY SOLD BY LEADING MARKET BRANDS IN INDIA

In this study, 133 milk samples (100 commercial liquid milk, 19 flavored milk and 14 milk powder) made from pasteurized milk by 10 leading commercial brands were purchased from the markets in Mathura and Agra districts of South Uttar Pradesh in North India. These milk samples were screened to estimate ‘bio-incidence’ of Mycobacterium avium subspecies paratuberculosis (MAP) using multiple tests; 3 antibody {Indigenous ELISA (i_ELISA), dot ELISA (d_ELISA) and Latex agglutination test (LAT)} and 3 antigen {(microscopy, Indirect fluorescent antibody test (i_FAT) and IS900 PCR}. Of 133 samples screened, 42.8, 58.6, 9.0, 27.0, 49.6 and 42.8% were positive for MAP in microscopy, i_FAT, IS900 PCR, i_ELISA, d_ELISA, and LAT, respectively. i_FAT was most sensitive followed by d_ELISA, LAT, microscopy, i_ELISA and IS900 PCR. In general, i_FAT, d_ELISA, LAT and microscopy were significantly superior to i_ELISA and IS900 PCR for estimating bio-incidence of MAP in milk samples. High bio-incidence of MAP in food items of mass consumption (liquid milk, flavoured milk and milk powder) made from pasteurized milk and freely sold in local markets by leading commercial brands emphasized the need for the immediate implementation of programs for the control of MAP in domestic livestock. MAP, the cause of incurable Johnes disease is endemic in * Corresponding author

Assessment of Ovine Johne’s disease in the Mandya sheep breed in South India using multiple diagnostic tests and bio-typing of Mycobacterium avium subspecies paratuberculosis infection

Abstract Johne’s disease is major infectious disease of domestic livestock in India. Despite low per animal productivity, the country is yet to initiate plan for the survey and control of disease at National scale. A total of 81 clinical samples from Mandya breed of sheep were collected from an organised farm (Livestock Research and Information Centre, Nagmangala) and farmer’s flocks suspected for Johne’s disease on the basis of clinical symptoms (weakness, emaciation, diarrhea and alopecia). Only 10.0% samples from two farms were screened for bio-load of Mycobacterium avium subspecies paratuberculosis (MAP) using multiple tests (microscopy, indigenous ELISA, IS900 PCR and culture). Results showed that 100.0% samples were positive for MAP infection both in microscopy and “Indigenous ELISA”. IS1311 PCR-REA bio-typed 60.0% (feces) and 100.0% (blood and tissue) isolates as “Indian Bison Type”. Typing investigated for the first time that biotype was of Indian origin and present in sheep flocks of South India having wide geographical distribution and broad host range. Study reported high bio-load of MAP in Mandya breed of sheep in Nagamangala, Mandya district and Bilagi taluk of Bagalkot district in South India. Being Johne’s disease a spectral disease, multiple tests were useful for the screening of MAP infection in sheep farms. Study emphasised the need for initiation of short and long term control programs to reduce production losses and sharing of MAP strains by other domestic livestock species and prevent contamination of human food chain and reduce risk to human population.