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This research analyzed the perceptions of veterinarians and farmers about paratuberculosis (PTB) and biosecurity practices in Pernambuco, a state region of Northeast Brazil. A questionnaire was administered to 84 veterinarians and 54 farmers with objective questions about the disease and control and prevention practices. Regarding knowledge about PTB, it was observed that 97.62% (n = 82/84) of veterinarians and 55.56% (n = 30/54) of farmers were aware of the disease (P < 0.0001). In connection with general biosecurity practices, it was found that 98.81% of veterinarians (n = 83/84) and 55.56% of farmers (n = 30/54) knew the term biosecurity and associated it with associated with measures to prevent the occurrence of occupational accidents (P < 0.05). The research data showed that the promotion of health education for these professionals about the PTB and biosecurity practices is an appropriate instrument to reduce direct and indirect costs caused by the disease and to promote the good health of herds in the state of Pernambuco, Northeast Brazil. RESUMO: Objetivou-se com essa pesquisa analisar a percepção de Médicos Veterinários e Produtores rurais sobre a paratuberculose (PTB) e as práticas de biossegurança no estado de Pernambuco da região Nordeste do Brasil. Aplicou-se um questionário para 84 veterinários e 54 produtores rurais com questões objetivas sobre a doença e as práticas de controle e prevenção. Em relação ao conhecimento sobre a paratuberculose, observou-se que 97,62% (n = 82/84) dos veterinários e 55,56% (n = 30/54) dos produtores rurais conheciam a doença (P < 0,0001). Em relação às práticas gerais de biossegurança, verificou-se que o termo biossegurança era conhecido por 98,81% dos veterinários (n = 83/84) e 55,56% dos produtores rurais (n = 30/54) associado às práticas de prevenção da ocorrência de acidentes de trabalho (P < 0,05). Os dados da pesquisa mostram que a promoção de educação em saúde para esses profissionais sobre a PTB e práticas de biosseguridade é um instrumento adequado para reduzir os custos diretos e indiretos causados pela doença e promover a boa saúde dos rebanhos no estado de Pernambuco.

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.

This research analyzed the perceptions of veterinarians and farmers about paratuberculosis (PTB) and biosecurity practices in Pernambuco, a state region of Northeast Brazil. A questionnaire was administered to 84 veterinarians and 54 farmers with objective questions about the disease and control and prevention practices. Regarding knowledge about PTB, it was observed that 97.62% (n = 82/84) of veterinarians and 55.56% (n = 30/54) of farmers were aware of the disease (P < 0.0001). In connection with general biosecurity practices, it was found that 98.81% of veterinarians (n = 83/84) and 55.56% of farmers (n = 30/54) knew the term biosecurity and associated it with associated with measures to prevent the occurrence of occupational accidents (P < 0.05). The research data showed that the promotion of health education for these professionals about the PTB and biosecurity practices is an appropriate instrument to reduce direct and indirect costs caused by the disease and to promote the good health of herds in the state of Pernambuco, Northeast Brazil. Key words: biosecurity, questionnaire, veterinarians, farmers, Johne disease. Objetivou-se com essa pesquisa analisar a percepção de Médicos Veterinários e Produtores rurais sobre a paratuberculose (PTB) e as práticas de biossegurança no estado de Pernambuco da região Nordeste do Brasil. Aplicou-se um questionário para 84 veterinários e 54 produtores rurais com questões objetivas sobre a doença e as práticas de controle e prevenção. Em relação ao conhecimento sobre a paratuberculose, observou-se que 97,62% (n = 82/84) dos veterinários e 55,56% (n = 30/54) dos produtores rurais conheciam a doença (P < 0,0001). Em relação às práticas gerais de biossegurança, verificou-se que o termo biossegurança era conhecido por 98,81% dos veterinários (n = 83/84) e 55,56% dos produtores rurais (n = 30/54) associado às práticas de prevenção da ocorrência de acidentes de trabalho (P < 0,05). Os dados da pesquisa mostram que a promoção de educação em saúde para esses profissionais sobre a PTB e práticas de biosseguridade é um instrumento adequado para reduzir os custos diretos e indiretos causados pela doença e promover a boa saúde dos rebanhos no estado de Pernambuco. Palavras-chave: biossegurança, questionário, veterinários, produtores rurais, doença de Johne.

•Gudair® vaccination is partially effective in reducing MAP faecal shedding.•The average animal level prevalence dropped from 7.64% to 0.12% after about 10years.•37.5% of the flocks were still infected after a decade of vaccination.•Clinical disease may recur if the program of annual lamb vaccinations is discontinued.•There is some risk of spread of MAP if vaccinated sheep are translocated. Since its registration in 2002, Gudair® vaccine has become the key tool for managing paratuberculosis in sheep in Australia. This study was conducted to evaluate its effectiveness in reducing paratuberculosis prevalence in 12 flocks over more than one decade. Flocks with variable initial prevalence (5, 4 and 3 with low, medium, and high prevalence, respectively) were enrolled in the study in 2003–04. Six biennial faecal samplings were conducted in these flocks over a 10year period. At each sampling, faeces from four age groups (3, 4, 5 and 6year-old sheep) were collected from each flock. The aim was to select 7 or 14 pools of faeces constituted with pellets from 50 or 25 sheep from each age group, respectively, but the numbers and sizes of pools did vary due to logistical issues. Samples were cultured using pooled faecal culture approach and sheep level paratuberculosis prevalence was estimated. Changes in probability of a pool being positive and in paratuberculosis prevalence over time were evaluated by fitting generalised-linear and linear mixed models. The proportion of positive faecal pools significantly declined over time from 50.3% at the first sampling in 2003–04 to only 3.1% at the last sampling in 2013–14, suggestive of a 30-fold reduction in the odds of a pooled faecal sample to be positive (p<0.001). Similarly, the average animal level prevalence in flocks dropped from 7.64% at the first sampling to 0.12% at the last sampling. However, faecal shedding persisted and was still present in 3 of the 8 flocks (37.5%) that remained in the study at the last sampling conducted in 2013–14. This is the longest study ever conducted to evaluate paratuberculosis vaccine effectiveness. The results will enable sheep producers to make informed decisions on managing paratuberculosis, and in evaluating the risks of purchasing and trading vaccinated sheep.

Mycobacterium avium subsp. paratuberculosis (MAP) is responsible for Johne’s disease (JD), a contagious granulomatous gastroenteritis with global distribution. This study investigated the expression of miRNAs and their potential regulatory mechanisms in Canadian Holstein cows with different MAP infection status, including 4 infected (MAPI), 5 tolerant to MAP infection (MAPT) and 5 healthy controls (HC). Intestinal tissues, jejunum (JE), jejunal lymph node (JELN) and caecal Peyer’s patches (CPP) were collected for miRNA-sequencing, differential expression and functional analysis. A total of 318, 281 and 251 miRNAs were identified in JE, JELN, and CPP, respectively. Out of these, 28, 46, and 24 were highly expressed in JE, JELN and CPP, respectively, including 23 miRNAs highly expressed across all tissues. Their functional enrichment indicates overrepresentation in mostly immune related functions. More differentially expressed (DE) miRNAs were identified in CPP (39, 21 and 11) followed by the JELN (25, 8 and 13) and JE (9, 9 and 15) for the comparisons MAPI vs HC, MAPT vs HC, and MAPI vs MAPT, respectively. Six common DE miRNAs (bta-miR-125a, bta-miR-146a, bta-miR-146b, bta-miR-21-5p, bta-miR-320a, bta-miR-370) (FDR < 0.1) in the JELN and CPP of MAPI cows are implicated in the immune response, suggesting roles in MAP infection. Similarly, more functional annotations were recorded for the CPP (1284) than JELN (377) and JE (four). DE miRNAs in JELN and CPP were enriched in gene ontology (GO) terms (e.g. lymphocyte activation, lymphocyte homeostasis, leukocyte differentiation, regulation of cell differentiation, T cell receptor complex, alpha–beta T cell activation) and KEGG pathways (e.g. T/B cell receptor signaling pathways, Chemokine signaling pathway, Leukocyte transendothelial migration, Th17 cell differentiation, MAPK signaling pathway, etc.) with immune related functions indicative of participation in the regulatory mechanisms of the host immune response to MAP infection. The comparison MAPI vs HC revealed a more pronounced immune response in JELN and CPP indicative of a heightened immune response, but fewer DE miRNAs and less pronounced immune activation for MAPT vs HC indicative of the development of tolerance to MAP presence. MiRNA expression and functional enrichment in JE, JELN, and CPP highlightes tissue-specific regulation of immune pathways and cellular functions during JD. Moreover, largely a different set of DE miRNAs, biological processes and pathways were driving the MAPI and MAPT phenotypes. These findings underscore the complex interplay between miRNAs and MAP in the studied tissues and provide a foundation for further exploration of miRNAs as potential biomarkers for the management of JD.

Paratuberculosis is a debilitating disease of ruminants that causes significant economic loss in both cattle and sheep. Early detection of the disease is crucial to controlling the disease; however, current diagnostic tests lack sensitivity. This study evaluated the potential for volatile organic compounds (VOCs) detected by gas chromatography and an electronic nose (eNose) for use as diagnostic tools to differentiate between Map-infected and non-infected cattle and sheep. Solid-phase micro-extraction gas chromatography–mass spectrometry (SPME GC-MS) was used to quantify VOCs from the headspace of faecal samples (cattle and sheep), and partial least squares–discriminant analysis (PLS-DA) was used to determine the suitability as a diagnostic tool. Both the cattle and sheep models had high specificity and sensitivity, 98.1% and 92.3%, respectively, in cattle, and both were 100% in sheep. The eNose was also able to discriminate between Map-infected and non-infected sheep and cattle with 88.9% specificity and 100% sensitivity in sheep and 100% specificity and sensitivity in cattle. This is the first time that VOC analysis by eNose and GCMS has been used for identification of Map in cattle and sheep faeces. GCMS also allowed the identification of putative disease biomarkers, and the eNose diagnostic capability suggests it is a promising tool for point-of-care diagnosis for Map detection on farms.

Background Paratuberculosis, caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a chronic granulomatous enteritis that affects domestic and wild ruminants and camelids. The disease has rarely been reported in alpacas in Germany. This publication describes epidemiologically independent cases of paratuberculosis in two alpacas in Germany. Case presentation Two alpacas, a 26-year-old female zoo animal (case 1) and a 2.5-year-old breeding stallion from a private owner (case 2), presented with progressive emaciation, leading to death (case 2) or euthanasia (case 1) because of deteriorating general condition. In both cases typical granulomatous lesions in the intestinal mucosa and mesenteric lymph nodes were found. In case 2, other lymph nodes were severely enlarged and MAP was detected in the mandibular lymph node, lung, and liver by qPCR. The MAP isolates differed between the alpacas, with two distinct phylogenetic clades (Clade 1 and 8) within Subgroup A of the MAP-C type group and two distinct INMV profiles (INMV 2 and 1) found. These genotypes have been identified in cattle and goats in different regions in Germany. The genotype isolated from case 1 has been detected in goats from the zoo since 2011, indicating transmission between these species. Conclusions MAP can cause severe clinical disease in alpacas of variable age and under different husbandry conditions. Therefore, paratuberculosis should be considered for differential diagnosis in alpacas with emaciation and poor general condition. Although not definitely shown, cross-species infection between ruminant species and camelids is exceedingly likely.

Current diagnostic methods for Johne’s disease in cattle allow reliable detection of infections with Mycobacterium avium ssp. paratuberculosis (MAP) not before animals are 2 years of age. Applying a flow cytometry-based approach (FCA) to quantify a MAP-specific interferon-gamma (IFN-γ) response in T cell subsets, the present study sought to monitor the kinetics of the cell-mediated immune response in experimentally infected calves. Six MAP-negative calves and six calves, orally inoculated with MAP at 10 days of age, were sampled every 4 weeks for 52 weeks post-inoculation (wpi). Peripheral blood mononuclear cells (PBMC) were stimulated with either purified protein derivatives (PPD) or whole cell sonicates derived from MAP (WCSj), M. avium ssp. avium or M. phlei for 6 days followed by labeling of intracellular IFN-γ in CD4 + and CD8 + T cells. No antigen-specific IFN-γ production was detectable in CD8 + cells throughout and the responses of CD4 + cells of MAP-infected and control calves were similar up to 12 wpi. However, the mean fluorescence intensity (MFI) for the detection of IFN-γ in CD4 + cells after WCSj antigen stimulation allowed for a differentiation of animal groups from 16 wpi onwards. This approach had a superior sensitivity (87.8%) and specificity (86.8%) to detect infected animals from 16 wpi onwards, i.e., in an early infection stage, as compared to the IFN-γ release assay (IGRA). Quantification of specific IFN-γ production at the level of individual CD4 + cells may serve, therefore, as a valuable tool to identify MAP-infected juvenile cattle.

subsp. (MAP) is the etiologic agent of Johne's disease in ruminants causing chronic diarrhea, malnutrition, and muscular wasting. Neonates and young animals are infected primarily by the fecal-oral route. MAP attaches to, translocates the intestinal mucosa, and is phagocytosed by macrophages. The ensuing host cellular immune response leads to granulomatous enteritis characterized by a thick and corrugated intestinal wall. We review various tissue culture systems, ileal loops, and mice, goats, and cattle used to study MAP pathogenesis. MAP can be detected in clinical samples by microscopy, culturing, PCR, and an enzyme-linked immunosorbent assay. There are commercial vaccines that reduce clinical disease and shedding, unfortunately, their efficacies are limited and may not engender long-term protective immunity. Moreover, the potential linkage with Crohn's disease and other human diseases makes MAP a concern as a zoonotic pathogen. Potential therapies with anti-mycobacterial agents are also discussed. The completion of the MAP K-10 genome sequence has greatly improved our understanding of MAP pathogenesis. The analysis of this sequence has identified a wide range of gene functions involved in virulence, lipid metabolism, transcriptional regulation, and main metabolic pathways. We also review the transposons utilized to generate random transposon mutant libraries and the recent advances in the post-genomic era. This includes the generation and characterization of allelic exchange mutants, transcriptomic analysis, transposon mutant banks analysis, new efforts to generate comprehensive mutant libraries, and the application of transposon site hybridization mutagenesis and transposon sequencing for global analysis of the MAP genome. Further analysis of candidate vaccine strains development is also provided with critical discussions on their benefits and shortcomings, and strategies to develop a highly efficacious live-attenuated vaccine capable of differentiating infected from vaccinated animals.

While paratuberculosis control has been studied for over a century, knowledge gaps still exist regarding the uptake and efficacy of control programmes. This narrative review aims to summarise studies on control programmes presented at the IDF ParaTB Fora in 2021 and 2022 and the International Colloquium on Paratuberculosis in 2022. Studies were grouped by topic as follows: successful control, field studies, education and extension, voluntary and compulsory control programmes, and surveillance. Various Map control programmes resulted in a decreasing animal and herd level Map prevalence. Long-term stakeholder commitment, stable funding, involvement of herd veterinarians and incentives for farmers to participate were shown to be pivotal for long-term success. Control measures focused on vertical and calf-to-calf transmission may improve Map control in infected herds. Easy-to-capture visualisation of surveillance test results to inform participants on the progress of Map control in their herds was developed. The probability of freedom from disease and estimated within-herd prevalence were identified as good candidates for categorisation of herds to support low-risk trade of cattle. Results of the surveillance schemes may inform genetic selection for resistance to Map infection. In conclusion, successful paratuberculosis control is feasible at both the herd and country level provided that crucial prerequisites are met.