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Abstract Background Tumor necrosis factor (TNF) is known to promote T cell migration and increase the expression of vascular endothelial growth factor (VEGF) and chemokines. The administration of Xpro-1595, a dominant-negative TNF (DN-TNF) engineered to selectively inactivate soluble TNF (solTNF), has been extensively studied and proven effective in reducing TNF production without suppressing innate immunity during infection. The literature also supports the involvement of glutamic acid-leucine-arginine (ELR+) chemokines and VEGF in angiogenesis and the spread of infections. Materials and methods In this study, we administered Xpro-1595 to guinea pigs to selectively inhibit solTNF, aiming to assess its impact on Mycobacterium tuberculosis (M.tb) dissemination, bacterial growth attenuation, and immunological responses. We conducted immunohistochemical analyses, immunological assays, and colony enumeration to comprehensively study the effects of Xpro-1595 by comparing with anti-TB drugs treated M.tb infected guinea pigs. Throughout the infection and treatment period, we measured the levels of Interleukin-12 subunit alpha (IL-12), Interferon-gamma (IFN-γ), TNF, Tumor growth factor (TGF), and T lymphocytes using ELISA. Results Our findings revealed a reduction in M.tb dissemination and inflammation without compromising the immune response during Xpro-1595 treatment. Notably, Xpro-1595 therapy effectively regulated the expression of VEGFA and ELR + chemokines, which emerged as key factors contributing to infection dissemination. Furthermore, this treatment influenced the migration of CD4 T cells in the early stages of infection, subsequently leading to a reduced T cell response and controlled proinflammatory signalling, thus mitigating inflammation. Conclusion Our study underscores the pivotal role of solTNF in the dissemination of M.tb to other organs. This preliminary investigation sheds light on the involvement of solTNF in the mechanisms underlying M.tb dissemination, although further in-depth research is warranted to fully elucidate its role in this process.

Abstract Background Detection of an appropriate antigen with high immunogenicity can be a big step in the production of an effective vaccine for control of Johne’s disease (JD). The aim of this study was to evaluate the efficacy of Mce-truncated protein as a subunit vaccine candidate for the control of JD in experimentally challenged goats. Materials and methods Six healthy goat kids were immunized with Mce-truncated protein, and two goats were kept as controls. All kids were twice challenged orally with live Mycobacterium avium subspecies paratuberculosis(MAP) strain and half the goats from both the categories were sacrificed at 7 and 10 months after start of challenge study. Culture of MAP was performed from all the necropsied tissues to determine the true JD infection status. Results Mce-truncated protein only reacted with pooled vaccinated goat sera in western-blot. A significant increase in humoral immune response against Mce protein was also observed in vaccinated goats. Compared to the control group, vaccinated goats gained higher body weights and none of them shed MAP or showed histopatological lesions or colonization of MAP in their necropsy tissues. Conclusions The new Mce protein based vaccine provided significant immunity in goats as they could meet the challenge with live MAP bacilli. Although the vaccine used in this study showed the high potential as a new effective vaccine for the control of JD, further validation study is still required to successfully implement the vaccine for JD control program.

Mycobacterium avium subspecies paratuberculosis (MAP) infection leads to chronic, persistent granulomatous enteritis, causing prolonged diarrhoea and emaciation. The disease is managed using medications such as antibiotics, live vaccines, mycobacteriophage therapies and other treatments; however, a notable proportion of affected animals do not show improvement with this approach. We hypothesise that immunoinhibitory receptors TIM-3 (T cell immunoglobulin mucin protein-3) and PD-1 (Programmed death receptor 1) may be upregulated on Peripheral blood mononuclear cells (PBMCs) of MAP-seropositive bovines, potentially contributing to immune exhaustion. Samples (blood and faeces) were collected from 32 diarrhoeic bovines suspected of MAP infection; eight apparently healthy buffaloes from the dairy farm at Hisar, Haryana and from 14 cows (suffering from chronic diarrhoea, weakness and emaciation) housed in stray cattle shed. MAP infection was estimated using indigenous ELISA (i-ELISA), faecal IS900 PCR, culture and acid-fast staining. TIM-3 and PD-1 gene expression on PBMCs were determined using qRT-PCR. TIM3 expression was relatively higher (~400-fold, 330-fold, 112-fold, 65-fold and 16-fold) in 5 chronically diarrhoeic PBMCs samples (MAP-seropositive), and higher PD-1 expression (around ~7-fold, 1.75-fold, 2.5-fold, 7.6-fold) was recorded in 4 diarrhoeic MAP-seropositive animals, compared to apparently healthy and other MAP-seronegative diarrhoeic animals. High co-expression of TIM-3 and PD-1 levels was also recorded in chronically diarrhoeic, emaciated stray cattle. Understanding immune responses in field conditions might aid in the therapeutic management of paratuberculosis.

Leprosy is a disease with spectral clinical manifestations along with two types of reactions, type 1 reaction (T1R) and type 2 reaction (T2R). T1R especially occurs because of the defensive upgradation of cell-mediated immunity (CMI) to M. leprae antigens. T1R is the main cause of disability in leprosy. The role of conventional adaptive T cells has been well studied to understand T1R. A comprehensive understanding of the role of unconventional T cells in the manifestation of inflammation during T1R is crucial and has not been studied. In our study, we found significantly higher plasma levels of TNFα, IL1β, IL17, and IP10 in T1R when compared to non-reaction (NR). Gene expression for cytokines in blood circulation by qPCR showed significantly higher expression of IFNγ, IP10, TNFα, IL6, IL17A and chemokines CCL3, CCR1, CCR5, and CXCR3 in T1R as compared to NR. Frequencies of NKT-like cells (48.7 %) and NK cells (22.3 %) were found significantly higher in T1R in comparison to NR (36.9 %, 18.3 %, respectively) (p = 0.0001). Significantly lower levels of γδT cells (3.32 %) were observed in T1R in comparison to NR (5.16 %). The present study has provided evidence for the first time on the role of plausible unconventional T cells in the immunopathogenesis of T1R in leprosy.

ObjectivesThe objective of this study is to analyze the association between TLR2 deletion (−196 to −174) and TLR1 743 A > G gene polymorphism with drug resistant tuberculosis (PTB, MDR-TB, and XDR-TB) in a population from Agra, Uttar Pradesh.MethodsThe present case–control study included 101 pulmonary TB patients, 104 multidrug-resistant TB patients, 48 extremely drug-resistant TB patients, and 130 healthy and unrelated controls residing in the same locality. The genotyping method for TLR2 deletion (−196 to −174) was carried out by allele-specific polymerase chain reaction (PCR), and TLR1 743 A > G gene polymorphism was performed by hybridization probe chemistry in Roche Real-Time PCR. Genotype and allele frequencies were analyzed by the chi-square test. Cytokine levels were measured by ELISA and compared using Mann–Whitney and Kruskal–Wallis tests.ResultsThe frequency of heterozygous (Ins/del) genotypes for TLR2 (−196 to −174) polymorphism was predominant in XDR-TB patients (0.57), whereas heterozygous A/G genotype for TLR1 743 A > G single nucleotide polymorphism (SNP) was predominant in healthy controls (0.57) for TLR1 743 A > G gene polymorphism. The heterozygous genotype of TLR2 deletion polymorphism was found to be significantly higher in XDR-TB (p = 0.0001). TLR1 743 A > G SNP, AG genotypes were found to be significantly associated with healthy controls than PTB (p = 0.047). The level of serum cytokines (IL-6, TNF-α, and IFN-γ) was also found to be significantly different among TB patients and healthy controls.ConclusionThe findings suggested that in the present population, the heterozygous (Ins/Del) genotype and deletion allele of TLR2 deletion (−196 to −174) polymorphism are associated with the risk for the development of drug-resistant TB. Furthermore, for TLR1 743 A > G gene polymorphism, A/G genotype, and G allele are found associated with healthy controls, suggesting the protective role against TB.

Introduction: Multiple mechanisms were proposed for the etiopathogenesis of type 1 diabetes mellitus (T1DM). An interplay between genetic, environmental, and immunological factors leads to generate autoantibodies against glutamic acid decarboxylase 65, intracellular adhesion (ICA)-512, and insulin autoantibodies (IAA) proteins of beta cells. Molecular mimicry is the best proposed mechanism of the above, but yet needs much solid evidence. Zinc transporter 8 (ZnT8) and proinsulin (PI) proteins of beta cells act as epitopes for some partially recognized proteins (MAP3865c, MAP1, 4αgbp, and MAP2404c) of mycobacterium paratuberculosis (MAP) generating autoantibodies against them. Similarly, other protein molecular mimickers were also reported. This review collects the latest evidence for the environmental etiopathogenesis of T1DM pertaining to MAP at the background of the phenomenon of molecular mimicry, and it proposes more directions for research in the field. Materials and Methods: Using the keywords “MAP and T1DM,” “MAP and zoonotic diseases,” T1DM and Molecular mimicry” and “T1DM and epitopes” searches were conducted in PubMed, Science Direct, and Google Scholar following PRISMA guidelines. This review related original articles and case reports were collected for the compilation of all proposed proteins having a tendency of molecular mimicry. This review includes 12 original articles and 17 case reports published till date. Results: A strong association was found in peptide pairs MAP3865c/ZnT8, 4αgbp/PI, and MAP2404c/PI between MAP and human beings. Discussion: Multiple MAP proteins show strong homology to human proteins generating antibodies against pancreatic beta cells. MAP exists in dairy products, posing a high risk of transmission to human beings. An uninterrupted but silent exposure of human beings to MAP could be an environmental trigger for T1DM.

Background: Despite of several guidelines metformin and glimepiride remains the first prescribed drug in most people with diabetes. Very often canagliflozin and teneligliptin remain the third prescribed OHA. Efficacy of teneligliptin and canagliflozin as solo drug or in combination with other single OHA’s on glycaemic parameters, lipid profiles & blood pressure is already established. But their efficacy on above parameters when used as third drug next to metformin and sulfonylurea is still not investigated. Aims and objectives: Aim of this study is to compare glycaemic and non-glycemic effects of canagliflozin and teneligliptin as third line drug. Objective is to establish their multiple pleotropic effects at this position. Methods: Subjects with uncontrolled glycaemic parameters with hypertension and diabetic dyslipidaemia already on half maximal doses of metformin and glimepiride were enrolled in the study. Out of 87 selections, 45 were prescribed canagliflozin 100 mg/d and to 42 subjects, teneligliptin 20mg/d, each for 24 weeks. Effects over HbA1c, fasting and post prandial plasma glucose, systolic and diastolic blood pressures, HDL’c and triglycerides were observed pre and post intervention. Results: As a third drug both canagliflozin and teneligliptin show significant reduction in HbA1c (1.5 versus 1.6%), fasting (-20.2 versus -28.8 mg %), and post prandial (-39.9 versus – 58.5mg %). Effects over systolic (4 mmHg versus 1.1 mmHg) and diastolic BP (3.1 mmHg versus 0.2 mmHg) was clinically and statistically significant (p = 0.3 and 0.1) respectively in both groups. A statistically insignificant (9.6 versus 4.1 mg%; p =) reduction in Triglycerides but significant (7.6 versus 1.2; p

Abstract Background This study aimed to screen the sera of goats and sheep from flocks suspected of Mycobacterium avium subsp. paratuberculosis (MAP) infection by a newly standardized Mce-truncated ELISA (Mt-ELISA) kit for the detection of antibodies against MAP. Four diagnostic applied tests were evaluated including Indigenous plate-ELISA (IP-ELISA), Mt-ELISA, fecal Polymerase Chain Reaction (f-PCR) and fecal culture (FC). Materials and methods Assuming the absence of a gold standard, latent-class models in a Bayesian framework were used to estimate the diagnostic accuracy of the four tests for MAP. Results Mt-ELISA had higher Sensitivity (Se) in sheep (posterior median: 0.68 (95% Probability Interval (PI): 0.43–0.95), while IP-ELISA recorded the highest Se in goats as 0.83 (95% PI, 0.61–0.97). The f-PCR Se estimate slightly differed between species [sheep 0.36 (0.19–0.58), goats 0.19 (0.08–0.35)], while the Se of FC was similar between species [sheep 0.29 (0.15–0.51), goats 0.27 (0.13–0.45)]. The specificity estimates for all tests were high, close to unity, and similar between species. Conclusion Overall, the results showed that the Mt-ELISA method can be used for MAP detection in small ruminants’ flocks.

Abstract Several Mycobacterial infections including leprosy and tuberculosis are known to evoke autoimmune responses by modulating homeostatic mechanism of the host. Presence of autoantibodies like, rheumatoid factor, anti-nuclear factor and antibodies to host, collagen, keratin, myelin basic protein (MBP) and myosin, have been earlier reported in leprosy patients. In the present study, we detected the role of mimicking epitopes between Mycobacterium leprae and host components in the induction of autoimmune response in leprosy. Based on our previous findings, we predicted and synthesized a total of 15 mimicking linear B cell epitopes (BCE) and 9 mimicking linear T cell epitopes (TCE) of keratin and MBP. Humoral and cell-mediated immune responses against these epitopes were investigated in Non-reaction (NR), Type 1 reaction (T1R) leprosy patients, and healthy controls. We observed significantly higher levels of antibodies against 8 BCE in T1R in comparison to NR leprosy patients. Further, we also found 5 TCE significantly associated with lymphocyte proliferation in the T1R group. Our results indicated that these epitopes play a key role in the induction of autoimmune response in leprosy and are also strongly associated with the inflammatory episodes of T1R. Conclusively, these molecules may be employed as a biomarker to predict the inflammatory episodes of T1R.

IntroductionImmunological reactions are frequent complications that may occur either before, during, or after treatment and affect 30–50% of leprosy patients. The presence of autoantibodies like rheumatoid factor, antinuclear factor, and antibodies to host collagen, keratin, actin, myosin, endothelial cells, and myelin basic protein (MBP) has been earlier reported in leprosy patients. The purpose of this study was to identify cross-reactive proteins in clinical samples such as saliva and slit skin scrapings (SSS) of leprosy patients which could be utilised as prognostic biomarkers for Type 1 Reaction (T1R) in leprosy.MethodA total of 10 leprosy patients in T1R and 5 healthy volunteers were recruited. The protein was extracted from their SSS and saliva samples, thereafter, isoelectric focusing (IEF) and two-dimensional PAGE were performed to analyse the proteins. Furthermore, the cross-reactivity was identified by western blotting host proteins in gel against purified IgG from Mycobacterium leprae soluble antigen (MLSA)- hyperimmunized rabbit sera, thereafter, cross-reactive proteins were identified by MS/MS. The cross-reactive host proteins were analysed for homologous bacterial proteins and B cell epitopes (BCEs) were predicted by using bioinformatic tools.ResultsA total of five spots of salivary proteins namely S100-A9, 35.3 kDa, and 41.5 kDa proteins, Serpin peptidase inhibitor (clade A), Cystatin SA-III, and four spots of SSS namely 41.4 kDa protein, Alpha-1 antitrypsin, vimentin, and keratin 1, were identified as cross-reactive. Further, a total of 22 BCEs of cross-reactive host proteins were predicted and visualised.DiscussionThis data provides strong evidence of cross-reactivity/molecular mimicry between host and pathogen in leprosy patients with reaction. These BCEs of cross-reactive proteins could be further studied to predict reactions and may be utilised as an early diagnostic biomarker for T1R in leprosy.

Mycobacterium avium subspecies paratuberculosis (MAP) is a chronic, contagious, and typically life-threatening enteric disease of ruminants caused by a bacterium of the genus Mycobacterium, but it can also affect non-ruminant animals. MAP transmission occurs through the fecal–oral pathway in neonates and young animals. After infection, animals generate IL-4, IL-5, and IL-10, resulting in a Th2 response. Early detection of the disease is necessary to avoid its spread. Many detection methods, viz., staining, culture, and molecular methods, are available, and numerous vaccines and anti-tuberculosis drugs are used to control the disease. However, the prolonged use of anti-tuberculosis drugs leads to the development of resistance. Whereas vaccines hamper the differentiation between infected and vaccinated animals in an endemic herd. This leads to the identification of plant-based bioactive compounds to treat the disease. Bioactive compounds of Ocimum sanctum and Solanum xanthocarpum have been evaluated for their anti-MAP activity. Based on the MIC50 values, Ursolic acid (12 µg/mL) and Solasodine (60 µg/mL) were found to be suitable for anti-MAP activity.

Mycobacterium avium subspecies paratuberculosis (MAP) infection in domestic livestock causes persistent diarrhea, weight loss, and death and is also a potential cause of Crohn’s disease (CD) in humans; notably, treatments against MAP are insufficient, costly, and can cause adverse reactions. Hence, plant-derived bioactive constituents have been taken into consideration in this regard. Herein, we present the results of two bioactive constituents (Solasodine and Ursolic acid) that were evaluated for their safety and efficacy against MAP protein (Dephospho-Coenzyme A kinase (DPCK) by utilizing in vitro assays and different tools of in silico biology. The ADME/t-test, the drug-likeness property test, pharmacophore modelling, and PASS prediction have proven that both the constituents have better binding capacities than the available antibiotic drugs used to target protein inhibition pathways. Through our observations, it can be inferred that these two phytochemicals can be adequately used to treat paratuberculosis, thereby combating inflammatory bowel disorders (IBD) of an autoimmune nature.

Mycobacterium avium subspecies paratuberculosis (MAP), being a dairy-borne pathogen, resistant of pasteurization and other sterilization techniques, is a major cause for development of inflammatory bowel disorders such as Johne’s disease (JD) in dairy animals and Crohn’s Disease (CD) in humans, for which no therapy is available to date. In the absence of effective therapy or a vaccine, management of CD has been accomplished by removal of the affected intestines. However, usually, even after removal of 2/3 of the intestine, CD reoccurs. Hence, there exists a need to develop an alternative therapy for such infection. The potential of herbals remains unexplored against MAP and related infections. Therefore, the conducted study is a novel initiative for the evaluation of anti-mycobacterial activity of bioactive extracts of Solanum xanthocarpum Schrad. & Wendl. against MAP infection. The said plant was authenticated according to the Ayurvedic Pharmacopoeia of India. Qualitative and quantitative evaluation of the extracts were done using chromatographic and spectroscopic techniques. Preliminary in vitro pharmacological assessments revealed the immunomodulatory and anti-inflammatory potential of the extracts. REMA assay was conducted to determine their anti-MAP activity along with determination of the best active extract. The hydro-alcoholic extract showed the best inhibition of MAP, providing a potential ray of hope against this emerging major pathogen of animals, and associated with Crohn’s disease and other autoimmune disorders in human beings.

Abstract Paratuberculosis, a chronic disease affecting ruminant livestock, is caused by Mycobacterium avium subsp. paratuberculosis (MAP). It has direct and indirect economic costs, impacts animal welfare and arouses public health concerns. In a survey of 48 countries we found paratuberculosis to be very common in livestock. In about half the countries more than 20% of herds and flocks were infected with MAP. Most countries had large ruminant populations (millions), several types of farmed ruminants, multiple husbandry systems and tens of thousands of individual farms, creating challenges for disease control. In addition, numerous species of free-living wildlife were infected. Paratuberculosis was notifiable in most countries, but formal control programs were present in only 22 countries. Generally, these were the more highly developed countries with advanced veterinary services. Of the countries without a formal control program for paratuberculosis, 76% were in South and Central America, Asia and Africa while 20% were in Europe. Control programs were justified most commonly on animal health grounds, but protecting market access and public health were other factors. Prevalence reduction was the major objective in most countries, but Norway and Sweden aimed to eradicate the disease, so surveillance and response were their major objectives. Government funding was involved in about two thirds of countries, but operations tended to be funded by farmers and their organizations and not by government alone. The majority of countries (60%) had voluntary control programs. Generally, programs were supported by incentives for joining, financial compensation and/or penalties for non-participation. Performance indicators, structure, leadership, practices and tools used in control programs are also presented. Securing funding for long-term control activities was a widespread problem. Control programs were reported to be successful in 16 (73%) of the 22 countries. Recommendations are made for future control programs, including a primary goal of establishing an international code for paratuberculosis, leading to universal acknowledgment of the principles and methods of control in relation to endemic and transboundary disease. An holistic approach across all ruminant livestock industries and long-term commitment is required for control of paratuberculosis.

Emerging antibiotic resistance among mycoplasma microorganisms is of major concern in present times as they cause various diseases in both animals and humans. Mycoplasmoses, infections caused by mycoplasma microorganisms have become common in recent past and have gained importance both due to inability to diagnose and difficulty to treat. Respiratory tract infection, mastitis, arthritis, and septicemia caused by Mycoplasma in livestock are responsible for causing heavy economic losses. These diseases are frequently reported from countries of Africa and Asia, including India. Antimycoplasma antibiotics are frequently being used as therapeutic agents for the treatment of mycoplasmoses infection in livestock. They include macrolides, tetracyclines, fluoroquinolones, and aminoglycosides which are the main antibiotic classes commonly used against mycoplasma globally. Oxytetracyclines are the commonest antibiotics used for decades followed by enrofloxacin, tylosin, and streptomycin. Danofloxacin, lincomycin, spiramycin, erythromycin, gamithromycin, azithromycin, clarithromycin, gentamicin, doxycycline, and tulathromycin are also used occasionally. Continuous and unregulated use of these antibiotics over prolonged period can lead to menace of antibiotic resistance which is aided by inappropriate doses and uncontrolled use. Resistance to some antibiotics is already emerging. Mycoplasmas have devised different resistance mechanisms for combating antimicrobial action of these drugs. Common mechanisms noted are acquisitions of proteins affecting ribosomal subunits, inhibition of antibiotic efflux, structural changes in the ribosomal subunit, target mutations, expression or production of enzymes. Additional novel mechanisms of resistance still need to be investigated. Strategies for prevention and encountering of this antibiotic resistance are being devised by alternating antibiotics in application, using antimycoplasma antibiotic sensitivity tests, along with evaluation of specific doses and exploration of novel mycoplasma specific class of antibiotics. Novel targets based on various cell structures including cell membrane, organelles, proteins, enzymes or metabolites are being explored for antimycoplasma therapy. These all will help in effective therapeutic management of mycoplasmoses with minimal side effects.

Contagious caprine pleuropneumonia (CCPP) is a serious disease of goats, occasionally sheep and wild ruminants, caused by Mycoplasma capricolum subspecies capripneumoniae (Mccp). The disease is characterized by severe serofibrinous pleuropneumonia, very high morbidity (∼100%), and mortality (80–100%). CCPP affects goats in more than 40 countries of the world thereby posing a serious threat to goat farming around the globe. The characteristic clinical signs of CCPP are severe respiratory distress associated with sero-mucoid nasal discharge, coughing, dyspnea, pyrexia, pleurodynia, and general malaise. In later stages, severe lobar fibrinous pleuropneumonia, profuse fluid accumulation in pleural cavity, severe congestion of lungs and adhesion formation is observed. Mycoplasmal antigen interactions with host immune system and its role in CCPP pathogenesis are not clearly understood. CCPP is not a zoonotic disease. Diagnosis has overcome cumbersome and lengthy conventional tests involving culture, isolation, and identification by advanced serological (LAT, cELISA) or gene-based amplification of DNA (PCR, RFLP, and hybridization) and sequencing. The latex agglutination test (LAT) is rapid, simple, and better test for field and real-time diagnosis applicable to whole blood or serum and is more sensitive than the CFT and easier than the cELISA. Moreover, the studies on antibiotic sensitivity and exploration of novel antibiotics (fluoroquinolones, macrolides) can help in better therapeutic management besides preventing menace of antibiotic resistance. Re-visiting conventional prophylactic measures focussing on developing novel strain-based or recombinant vaccines using specific antigens (capsular or cellular) should be the most important strategy for controlling the disease worldwide.

High endemicity of Johne’s disease (JD) in herds adversely affects heavy milk yielding breeds by reducing the per animal productivity and ‘productive life-span’. This review evaluates different vaccines used for its control and summarizes the benefits of ‘global vaccine’ in the four major domestic livestock species, namely goat, sheep, buffalo and cattle. Vaccines developed by using ‘native strains’ revealed both 'therapeutic' and preventive effects in domestic livestock. The 'therapeutic' role of vaccine in animals suffering from clinical JD turned out to be valuable in some cases by reversing the disease process and animals returning back to health and production. Good herd management, improved hygiene, ‘test and cull’ methodology, proper disposal of animal excreta and monitoring of MAP bio-load were also regarded as crucial in the 'therapeutic' management of JD. Vaccine approaches have been widely adopted in JD control programs and may be considered as a valuable adjunct in order to utilize huge populations of otherwise un-productive livestock. It has been shown that vaccination was the preeminent strategy to control JD, because it yielded approximately 3–4 times better benefit-to-cost ratios than other strategies. Internationally, 146 vaccine trials/studies have been conducted in different countries for the control of JD and have shown remarkable reduction in its national prevalence. It is concluded that for JD, there cannot be global vaccines or diagnostic kits as solutions have to come from locally prevalent strains of MAP. Despite some limitations, vaccines might still be an effective strategy to reduce or eradicate JD.

Present study is based on 24 goats that were located in goat herds endemically infected with Mycobacterium avium subspecies paratuberculosis (MAP) infection. Objective of the study was to access the variations in the lipid profile and acute phase proteins in the serum samples driven from non-infected (negative and physically healthy) and infected (positive and physically weak) goats with MAP infection, cause of incurable Johne’s disease (JD) in domestic livestock. Infected goats had significantly higher cholesterol and albumin levels and significantly ‘reduced level’ of high density lipoprotein (HDL) and ‘reduced level’ of the density of lipoproteins (LDL) in comparison to non-infected goats. Lipid profile and acute phase proteins could be further explored for their significance in pathogenesis and diagnosis of JD in domestic livestock including goats.

Johne's disease (JD) is an infectious wasting condition of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP) in domestic livestock of every country that has been investigated. Controlling JD is problematic due to the lack of sensitive, specific, efficient, and cost-effective diagnostic tests. A major challenge in the development of diagnostics like ELISA is the selection of an ideal antigen/(s) that is pathogen-specific and allows sensitive recognition. Therefore, the purpose of this study was to identify and use Mce-truncated protein-based ELISA assay for the diagnosis of MAP infection with high sensitivity and specificity. In silico epitope prediction by epitope mapping throughout the whole length of MAP2191 protein revealed that C-terminal portion of this protein presented potential T- and B-cell epitopes. Therefore, a novel Mce-truncated protein encoded by the selected region of MAP2191 gene was expressed, purified with Ni-NTA gel matrix and confirmed by SDS PAGE and western blot. A profiling ELISA assay was developed to evaluate sera from MAP infected and non-infected ruminant species for antibodies against Mce-truncated protein to infer the immunogenicity of this protein in the host. Using this Mce protein-based ELISA, 251 goats, 53 sheep, 117 buffaloes, and 33 cattle serum samples were screened and 49.4, 51.0, 69.2, and 54.6% animals, respectively, were found positive. Comparing with i-ELISA, the new Mce-based ELISA kit showed a relatively higher specificity but suffered from slightly reduced sensitivity. Mce-based ELISA excluded apparently false positive results of i-ELISA. Mce protein was found to be antigenic and Mce-ELISA test could be employed as a diagnostic test for JD in domestic livestock in view of the a relatively higher specificity and accuracy. The antigenic potential of Mce antigen can also be exploited for the development of a new vaccine for the control of MAP infection.

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.

Sarco/endoplasmic reticulum calcium-ATPase (SERCA) is a membrane-bound cytosolic enzyme which 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 virus replication. Treatment of Vero cells with SERCA-specific inhibitor (Thapsigargin) at a concentration that is nontoxic to the cells significantly reduced Peste des petits ruminants virus (PPRV) and Newcastle disease virus (NDV) replication. Conversely, overexpression of SERCA rescued the inhibitory effect of Thapsigargin on virus replication. PPRV and NDV infection induced SERCA expression in Vero cells, which could be blocked by Thapsigargin. Besides inducing enhanced formation of cytoplasmic foci, Thapsigargin was shown to block viral entry into the target cells as well as synthesis of viral proteins. Furthermore, NDV was shown to acquire significant resistance to Thapsigargin upon long-term passage (P) in Vero cells. As compared to the P0 and P70-Control, the fusion (F) protein of P70-Thapsigargin virus exhibited a unique mutation at amino acid residue 104 (E104K), whereas no Thapsigargin-associated mutations were observed in HN gene. To the best of our knowledge, this is the first report describing the 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.

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.

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.

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 one 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.

Johne’s disease of domestic livestock has high economic significance. Environmental factors and farm level management practices are associated with the incidence and occurrence of disease in farm and farmers herds/ flocks. A cross-sectional study was conducted in the dairy herds (315) maintained in different geographical regions and management practices in the Punjab state to determine ‘herd level’ risk factors associated with Johne’s disease. Of 16 factors studied, univariate analysis showed that 6 factors were significantly associated with sero-positivity. Multivariate analysis showed contamination of feed and water with adult manure (OR=3.97) and history of chronic diarrhoea in the herd (OR=2.04) as the factors significantly associated with positive status of animals in the herd. It is the first report on ‘risk factors’ analysis for Johne’s disease in India.