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novel monoclonal antibody and peptide binders for mycobacterium avium subsp. paratuberculosis and their application for magnetic separation.the generation of novel mycobacterium avium subsp. paratuberculosis (map)-specific monoclonal antibodies and phage-display derived peptide binders, along with their application for the magnetic separation (ms) of map cells, is described. our aim was to achieve even greater map capture capability than is possible with peptide-mediated magnetic separation (pms) using a 50:50 mix of biotinylated-amp3 and biotinylated-amptd peptide-coated beads. gamma-irradiated whole map cells and ethanol extracted ...201626815790
mycobacterial infections in rabbits: from the wild to the laboratory.tuberculous mycobacterial diseases such as leprosy and tuberculosis are ancient diseases that currently continue threatening human health in some countries. non-tuberculous mycobacterial (ntm) infections cause a series of well-defined pathological entities, as well as some opportunistic diseases that have also increased worldwide, being more common among immunocompromised patients but rising also in immunocompetent individuals. reports on natural infections by mycobacteria in rabbits are scarce ...201626799551
dam's infection progress and within-herd prevalence as predictors of mycobacterium avium subsp. paratuberculosis elisa response in danish holstein cattle.understanding the primary routes of transmission of mycobacterium avium subsp. paratuberculosis (map) is pivotal to manage the pathogen in cattle herds. map is transmitted both vertically and horizontally, and both the dam's stage of infection and the prevalence in the population are therefore potentially important for map transmission control. the objective of this study was therefore to assess the dam's infection progress and the within-herd test-prevalence as predictors of map infection in da ...201626795463
correction: analysis of biobanked serum from a mycobacterium avium subsp paratuberculosis bovine infection model confirms the remarkable stability of circulating mirna profiles and defines a bovine serum mirna repertoire. 201626785077
bayesian estimation of sensitivity and specificity of a commercial serum/milk elisa against the mycobacterium avium subsp. paratuberculosis (map) antibody response for each lactation stage in greek dairy sheep.a total of 854 paired milk and blood samples were collected from ewes of a greek flock and used to estimate the sensitivity and specificity of a commercial elisa for detection of mycobacterium avium subsp. paratuberculosis (map) specific antibodies in each stage of lactation. we implemented bayesian mixture models to derive the distributions of the test response for the healthy and the infected ewes. in the colostrum period, early, mid and late lactation stage the median values of the area under ...201626754926
modeling of mycobacterium avium subsp. paratuberculosis dynamics in a dairy herd: an individual based approach.in the dairy industry, johne's disease (jd), caused by mycobacterium avium subsp. paratuberculosis (map) is one of the major investigated diseases. to date, researchers have suggested some control strategies for jd, such as test-and-cull based herd management, isolated calf rearing management, and vaccinations. due to the slow progressing nature of map, tests with low diagnostic test sensitivity and specificity, and economic limitations, implementing these strategies has not resulted in eliminat ...201627521525
dysbiosis of the fecal microbiota in cattle infected with mycobacterium avium subsp. paratuberculosis.johne's disease (jd) is a chronic, intestinal infection of cattle, caused by mycobacterium avium subsp. paratuberculosis (map). it results in granulomatous inflammation of the intestinal lining, leading to malabsorption, diarrhea, and weight loss. crohn's disease (cd), a chronic, inflammatory gastrointestinal disease of humans, has many clinical and pathologic similarities to jd. dysbiosis of the enteric microbiota has been demonstrated in cd patients. it is speculated that this dysbiosis may co ...201627494144
development of vaccines to mycobacterium avium subsp. paratuberculosis infection.johne's disease or paratuberculosis is a chronic debilitating disease in ruminants caused by mycobacterium avium subsp. paratuberculosis (map). the disease causes significant economic losses in livestock industries worldwide. there are no effective control measures to eradicate the disease because there are no appropriate diagnostic methods to detect subclinically infected animals. therefore, it is very difficult to control the disease using only test and cull strategies. vaccination against par ...201627489800
marked differences in mucosal immune responses induced in ileal versus jejunal peyer's patches to mycobacterium avium subsp. paratuberculosis secreted proteins following targeted enteric infection in young calves.in cattle, mycobacterium avium subsp. paratuberculosis infection is primarily mediated through m cells overlying peyer's patches (pp) in the ileum. the capacity of m. avium subsp. paratuberculosis to invade ileal pp (ipp) versus discrete pp in the jejunum (jpp) and subsequent differences in mucosal immune responses were investigated. intestinal segments were surgically prepared in both mid-jejunum, containing two jpps, and in terminal small intestine containing continuous ipp. m. avium subsp. pa ...201627387969
a rapid method for quantifying viable mycobacterium avium subsp. paratuberculosis in cellular infection assays.determining the viability of bacteria is a key outcome of in vitro cellular infection assays. currently, this is done by culture, which is problematic for fastidious slow-growing bacteria such as mycobacterium avium subsp. paratuberculosis, where it can take up to 4 months to confirm growth. this study aimed to identify an assay that can rapidly quantify the number of viable m. avium subsp. paratuberculosis cells in a cellular sample. three commercially available bacterial viability assays along ...201627371585
humoral response against host-mimetic homologous epitopes of mycobacterium avium subsp. paratuberculosis in japanese multiple sclerosis patients.several works have demonstrated the existence of a link between mycobacterium avium subsp. paratuberculosis (map) and ms in italy. in this study, we analyzed the serology of map in a japanese population while looking at several markers of map. fifty ms patients, 12 clinically isolated syndrome (cis) patients, 30 other neurological disorders (ond) patients, and 50 healthy controls (hcs) were tested using elisa for the presence of igg antibodies toward immunodominant epitopes map_0106c121-132, hom ...201627356622
seroreactivity against specific l5p antigen from mycobacterium avium subsp. paratuberculosis in children at risk for t1d.although numerous environmental agents have been investigated over the years as possible triggers of type 1 diabetes (t1d), its causes remain unclear. we have already demonstrated an increased prevalence of antibodies against peptides derived from mycobacterium avuim subsp. paratuberculosis (map) homologous to human zinc transporter 8 protein (znt8) and proinsulin in italian subjects at risk for or affected by t1d. in this study, we compared titers of the previously detected antibodies with sero ...201627336739
paratuberculosis in buffaloes in northeast brazil.several farms in the northeast of brazil were investigated for mycobacterium avium subsp. paratuberculosis infection in order to identify the occurrence of paratuberculosis in buffaloes. samples were obtained from 17 farms, two slaughter houses, and a quarantine area in the northeast. about 15,000 buffaloes of the murrah, mediterranean, and jafarabadi breed as well as their crossbreeds were evaluated for meat, dairy, and mixed farms with semi-intensive or extensive breeding practices. for diagno ...201627334632
evaluation of the limitations and methods to improve rapid phage-based detection of viable mycobacterium avium subsp. paratuberculosis in the blood of experimentally infected cattle.disseminated infection and bacteraemia is an underreported and under-researched aspect of johne's disease. this is mainly due to the time it takes for mycobacterium avium subsp. paratuberculosis (map) to grow and lack of sensitivity of culture. viable map cells can be detected in the blood of cattle suffering from johne's disease within 48 h using peptide-mediated magnetic separation (pmms) followed by bacteriophage amplification. the aim of this study was to demonstrate the first detection of m ...201627305900
pcr-based detection of mycobacterium avium subsp. paratuberculosis infection in cattle in south korea using fecal samples.mycobacterium avium subsp. paratuberculosis (map) is the causative agent of bovine paratuberculosis (ptb). the first step in the control of ptb is the identification and isolation of sub-clinical fecal shedders from the herd. in the current study, real-time and nested pcr targeting map-specific genetic elements (is900 and ismap02) dna isolated from fecal samples were used to detect map infection in cattle. of the 1,562 fecal samples obtained from 37 herds, regardless of diarrhea, 35 samples test ...201627301582
paratuberculosis in sheep: histochemical, immunohistochemical and in situ hybridization evidence of in utero and milk transmission.to investigate in utero and milk transmission of mycobacterium avium subsp. paratuberculosis (map), tissues from thirteen pregnant sheep, naturally infected and serologically positive to map, were examined by means of histochemistry, immunohistochemistry and in situ hybridization. soon after parturition, ewes were euthanized and tissues samples were collected and prepared. the offspring (18 lambs) were divided into three groups to investigate different routes of map transmission. lambs were sacr ...201627234557
studies on pathology, cytokine gene expression and molecular typing of mycobacterium avium subsp. paratuberculosis of naturally occurring johne's disease in bullocks.pathology of johne's disease (jd) in bullocks (castrated, adult male cattle) is rarely studied. here, we report the pathology and cytokine gene expression of naturally occurring jd in bullocks. the small intestine and mesenteric lymph nodes collected from 404 bullocks, aged between 5 and 10years, were examined for jd lesions and detection of mycobacterium avium subsp. paratuberculosis (map). a total of 8.7% bullocks exhibited jd lesions, which were classified into multibacillary-diffuse granulom ...201627234539
the genomics of mycobacteria.the species mycobacterium bovis and mycobacterium avium subspecies paratuberculosis are the causal agents, respectively, of tuberculosis and paratuberculosis in animals. both mycobacteria, especially m. bovis, are also important to public health because they can infect humans. in recent years, this and the impact of tuberculosis and paratuberculosis on animal production have led to significant advances in knowledge about both pathogens and their host interactions. this article describes the cont ...201627217180
milk yield and lactation stage are associated with positive results to elisa for mycobacterium avium subsp. paratuberculosis in dairy cows from northern antioquia, colombia: a preliminary study.paratuberculosis is a slow-developing infectious disease characterized by chronic granulomatous enterocolitis. this disease has a variable incubation period from 6 months to over 15 years and is caused by mycobacterium avium subsp. paratuberculosis (map). some studies have been conducted in cattle during the last decades in colombia. however, those studies were designed using relatively small populations and were not aimed to establish prevalence. this study aimed to determine the map seropreval ...201627165342
corrigendum to "a new compartmental model of mycobacterium avium subsp. paratuberculosis infection dynamics in cattle" [prev. vet. med. 122 (3) (2015) 298-305]. 201627143493
immunological findings associated with argentinean strains of mycobacterium avium subsp. paratuberculosis in bovine models.mycobacterium avium subspecies paratuberculosis (map) is the causative agent of ruminant paratuberculosis. the aim of this study was to evaluate the biological behavior of different argentinean strains of map in two bovine infection models: macrophage (in vitro) and calf (in vivo) through the evaluation of early immune responses at the peripheral and local levels. two map strains (a and c) were selected taking into account the different patterns of tnf-α and il-10 secretion displayed by infected ...201627138443
faecal bacterial composition in dairy cows shedding mycobacterium avium subsp. paratuberculosis in faeces in comparison with nonshedding cows.the aim of this study was to determine possible differences in the faecal microbiota of dairy cows infected with mycobacterium avium subsp. paratuberculosis (johne's disease) in comparison with noninfected cows from the same herds. faecal samples from cows in 4 herds were tested for m. avium subsp. paratuberculosis by real-time pcr, and faecal bacterial populations were analysed by 454 pyrosequencing of the 16s rrna gene. the most notable differences between shedding and nonshedding cows were an ...201627127920
altered microrna expression and pre-mrna splicing events reveal new mechanisms associated with early stage mycobacterium avium subspecies paratuberculosis infection.the molecular regulatory mechanisms of host responses to mycobacterium avium subsp. paratuberculosis (map) infection during the early subclinical stage are still not clear. in this study, surgically isolated ileal segments in newborn calves (n = 5) were used to establish in vivo map infection adjacent to an uninfected control intestinal compartment. rna-seq was used to profile the whole transcriptome (mrnas) and the micrornaome (mirnas) of ileal tissues collected at one-month post-infection. the ...201627102525
johne's disease: reliability of environmental sampling to characterize mycobacterium avium subspecies paratuberculosis (map) infection in beef cow-calf herds.environmental samples are considered to be a cost-effective method of identifying mycobacterium avium subsp. paratuberculosis (map)-positive dairy herds, but evidence for beef cow-calf herds is weak. this study aims at evaluating this approach in a total of 20 german herds that were characterized by individual faecal samples (n = 2545) of all cows. for 14 map-positive herds having at least one map-positive animal, the within-herd prevalence was calculated from concurrent individual faecal cultur ...201627094619
mesenteric lymph node granulomatous lesions in naturally infected wild boar (sus scrofa) in portugal--histological, immunohistochemical and molecular aspects.several studies have demonstrated that wildlife reservoirs of mycobacteria are responsible for the maintenance and spreading of the infection to livestock and wildlife counterparts. recent data report the role of wild boar (sus scrofa) as a reservoir for mycobacterium bovis. this study was conducted to evaluate the chronic inflammatory response in the mesenteric lymph nodes (mln) of wild boar with granulomatous lymphadenitis (n=30). morphological parameters of the lesions were recorded. the expr ...201627090622
mycobacterium avium subspecies paratuberculosis infection modifies gut microbiota under different dietary conditions in a rabbit model.mycobacterium avium subspecies paratuberculosis (map) the causative agent of paratuberculosis, produces a chronic granulomatous inflammation of the gastrointestinal tract of ruminants. it has been recently suggested that map infection may be associated with dysbiosis of intestinal microbiota in ruminants. since diet is one of the key factors affecting the balance of microbial populations in the digestive tract, we intended to evaluate the effect of map infection in a rabbit model fed a regular o ...201627065994
is there a role for mycobacterium avium subspecies paratuberculosis in parkinson's disease?in parkinson's disease (pd) znt proteins play an important role. zinc is a co-factor of numerous enzymes and stabilizes the tertiary structure of several proteins. nothing is known about previous infections mediated by mycobacterium avium subsp. paratuberculosis (map). we evaluated if a previous infection with map could induce the production of antibodies that cross-reacted with the znt homologous antigenic peptides associated to parkinson. the humoral response toward map3865c peptides, znt3 and ...201627049567
gene expression profiles of putative biomarker candidates in mycobacterium avium subsp. paratuberculosis-infected cattle.this study was conducted to analyze the gene expression of prognostic potential biomarker candidates using the whole blood of cattle naturally infected with italic! mycobacterium aviumsubsp. italic! paratuberculosis(map). we conducted real-time pcr to evaluate 23 potential biomarker candidates. experimental animals were divided into four groups based on fecal map pcr and serum elisa. seven ( italic! klrb1, italic! hgf, italic! mpo, italic! ltf, italic! serpine1, italic! s100a8and italic! s100a9) ...201627029383
serum and tissue concentrations of gallium after oral administration of gallium nitrate and gallium maltolate to neonatal calves.to determine serum and tissue concentrations of gallium (ga) after oral administration of gallium nitrate (gan) and gallium maltolate (gam) to neonatal calves.201627027708
presence of mycobacterium avium subsp. paratuberculosis in alpacas (lama pacos) inhabiting the chilean altiplano.mycobacterium avium subsp. paratuberculosis (map) is the etiologic agent of paratuberculosis. the organism causes disease in both domestically managed and wild ruminant species. south american camelids have a long, shared history with indigenous people in the andes. over the last few decades, increasing numbers of alpacas were exported to numerous countries outside south america. no paratuberculosis surveillance has been reported for these source herds. in this study, individual fecal samples fr ...201627010259
the isolation and molecular characterization of mycobacterium avium subsp. paratuberculosis in shandong province, china.mycobacterium avium subspecies paratuberculosis (map) causes johne's disease in domestic and wild ruminants. it has been a debate that whether map can cause crohn's disease in human. to our knowledge there is no report about molecular characterization of map in china, although several map strains have been reported in other country. the objectives of this study was to know the recent prevalence of johne's disease in dairy farms in shandong province, and have a better understanding of genotypic d ...201627006704
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. ...201627833911
first report of miru-vntr genotyping of mycobacterium avium subsp. paratuberculosis isolates from egypt.mycobacterium avium subsp. paratuberculosis (map) is the causative agent of johne's disease, an economically important disease in ruminants worldwide. it was first isolated in egypt in 2005. since then, the pathogen has been detected in different egyptian provinces. in order to trace the source of infection, genotyping using simple methods of high discriminatory power such as mycobacterial interspersed repetitive unit-variable number tandem repeats (miru-vntr) were carried out in different count ...201627822239
a single capsule formulation of rhb-104 demonstrates higher anti-microbial growth potency for effective treatment of crohn's disease associated with mycobacterium avium subspecies paratuberculosis.most recently we reported that rhb‑104 triple antibiotics combination in culture is bactericidal and should be effective for treatment of crohn's disease (cd)-associated with mycobacterium avium subspecies paratuberculosis (map) (alcedo et al. in gut pathog 14:32, 2016). the combination exhibited unique synergistic antimicrobial growth activity. the proprietary rhb-104 capsule formulation contains active ingredients (63.3 % clarithromycin (cla), 6.7 % clofazimine (clo) and 30 % rifabutin (rif)). ...201627708718
rhb-104 triple antibiotics combination in culture is bactericidal and should be effective for treatment of crohn's disease associated with mycobacterium paratuberculosis.mycobacterium avium subspecies paratuberculosis (map) has been implicated as an etiological agent of crohn's disease (cd), a debilitating chronic inflammatory bowel disease. clarithromycin (cla), clofazimine (clo), rifabutin (rif) and other antibiotics have been used individually or in combinations with other drugs to treat mycobacterial diseases including cd. the treatment has varied by regimen, dosage, and duration, resulting in conflicting outcomes and additional suffering to the patients. rh ...201627307791
oxidative stress due to mycobacterium avium subspecies paratuberculosis (map) infection upregulates selenium-dependent gpx activity.this study was designed to determine the relationship between mycobacterium avium subspecies paratuberculosis (map) infection and selenium-dependent glutathione peroxidase (gpx) activity, in the blood of humans and cattle infected with map.201626997979
comparative evaluation of diagnostic tests for the detection of mycobacterium avium subsp. paratuberculosis in the tissues of sheep affected with distinct pathology of paratuberculosis.paratuberculosis or johne's disease is a chronic infectious granulomatous enteritis, mainly of cattle, sheep, goats, and other domestic and wild animals caused by mycobacterium avium subsp. paratuberculosis (map). currently, map has been recognized as an important animal pathogen with significant zoonotic and public health concerns. the early detection of infected animals using suitable diagnostic methods helps in developing control and preventive strategies for the herd. therefore, the present ...201628043635
mycobacterial coinfection and persisting bovine tuberculosis-has the time arrived for a policy review?bovine tuberculosis (btb) is mainly a disease of cattle, although it continues to infect human populations across the world. operation of a test and slaughter plan in iran since 1981 has lowered the frequency of btb from >5% to <0.14% at the national scale. in 2015, unusual uncontrollable epidemics of btb were detected in two cattle farms in municipal suburbs of qazvin and isfahan. these farms had a tuberculin-test-certified record of btb-free status for the past 5 consecutive years, with no new ...201628043632
isolation and molecular identification of mycobacterium from commercially available pasteurized milk and raw milk samples collected from two infected cattle farms in alborz province, iran.mycobacterium avium subsp. paratuberculosis (map) is an etiological agent of johne's disease in ruminant including cattle, sheep and goats. this disease is considered an economically important disease in cattle. animals with paratuberculosis shed viable map, particularly in their milk and feces. map may be involved in the development of crohn's disease in humans through the consumption of contaminated milk and dairy products. common methods of pasteurization are not enough to kill all map presen ...201628043568
detection of mycobacterium avium subsp. paratuberculosis in cattle by using indirect absorbed elisa (enzyme-linked immunosorbent assay) system and culture in alborz province, iran.mycobacterium avium subsp. paratuberculosis is a slow growing, gram-positive, and acid-fast bacillus. it is the causative agent of the chronic enteritis disease of ruminants called johne's disease. most of the infected animals are young. however, most clinical cases belong to adult cattle aged 3-5years. due to a prolonged incubation period, identification of subclinically infected animals is one of the most crucial problems in johne's disease. the aim of this study was to detect m. avium subsp. ...201628043567
pathological, bacteriological, and molecular characteristics of natural outbreaks of johne's disease in goats of fars province, iran.johne's disease, also called paratuberculosis, is a very important chronic infectious disease of ruminants worldwide. the causative agent is an acid-fast bacterium, mycobacterium avium paratuberculosis (map). finding a precise method for detecting map is essential for the control and eradication of this disease in small ruminant herds.201628043554
development of an indirect elisa system to diagnosis paratuberculosis using surface antigens of mycobacterium avium subsp. paratuberculosis in cattle.mycobacterium avium subsp. paratuberculosis (map) is the causative agent of paratuberculosis (ptb), also called as johne's disease, and is considered as the cause of irreparable economic losses in livestock industry. this bacterium is excreted in large amounts through the milk and feces of infected animal. for the detection of the ptb, indirect elisa has been highly considered as a simple method with high sensitivity and specificity. accordingly, this study aims at designing a system of indirect ...201628043544
the structure-antimicrobial activity relationships of a promising class of the compounds containing the n-arylpiperazine scaffold.this research was focused on in silico characterization and in vitro biological testing of the series of the compounds carrying a n-arylpiperazine moiety. the in silico investigation was based on the prediction of electronic, steric and lipohydrophilic features. the molecules were screened against mycobacterium avium subsp. paratuberculosis cit03, m. smegmatis atcc 700084, m. kansasii dsm 44162, m. marinum camp 5644, staphylococcus aureus atcc 29213, methicillin-resistant s. aureus 63718, escher ...201627681720
serum baff levels, methypredsinolone therapy, epstein-barr virus and mycobacterium avium subsp. paratuberculosis infection in multiple sclerosis patients.elevated b lymphocyte activating factor baff levels have been reported in multiple sclerosis (ms) patients; moreover, disease-modifying treatments (dmt) have shown to influence blood baff levels in ms patients, although the significance of these changes is still controversial. in addition, baff levels were reported increased during infectious diseases. in our study, we wanted to investigate on the serum baff concentrations correlated to the antibody response against mycobacterium avium subspecie ...201627383531
epstein barr virus and mycobacterium avium subsp. paratuberculosis peptides are recognized in sera and cerebrospinal fluid of ms patients.mycobacterium avium subsp. paratuberculosis (map) and epstein-barr virus (ebv) epitopes elicit a consistent humoral response in serum of multiple sclerosis patients, but the cross reactivity against the homologous myelin basic protein (mbp) and human interferon regulatory factor 5 (irf5) has not been searched within the cerebral spinal fluid (csf). we evaluated in sera and csf of patients with ms and with other neurological diseases (ond) the humoral response against ebv/map peptides and the irf ...201626956729
immune response induced by epstein-barr virus and mycobacterium avium subsp. paratuberculosis peptides in current and past infectious mononucleosis: a risk for multiple sclerosis?infectious mononucleosis (im) caused by epstein-barr virus (ebv) has been associated with increased risk of multiple sclerosis (ms). however, the mechanism linking these pathologies is unclear. different reports indicate the association of ebv, and recently mycobacterium avium subsp. paratuberculosis (map), with ms. for a better understanding of the role of these pathogens, the host response induced by selected antigenic peptides in subjects with a history of im that significantly increases the ...201626453465
composition and potency characterization of mycobacterium avium subsp. paratuberculosis purified protein derivatives.mycobacterium avium subsp. paratuberculosis (map) purified protein derivatives (ppds) are immunologic reagents prepared from cultured filtrates of the type strain. traditional production consists of floating culture incubation at 37°c, organism inactivation by autoclaving, coarse filtration, and protein precipitation. three traditional production ppds were used in this study including lot 9801, which served as a reference and has been used in the field for decades. alternative production ppds (0 ...201627136199
expression and purification of a gene encoding a 9.7 kda pe protein of mycobacterium avium subsp. paratuberculosis.mycobacterium avium subsp. paratuberculosis (map) contains pe family antigens which are proline and glutamic acid rich and may play important role as t-cell antigens. in the present study, the map 1507 orf encoding 9.7 kda pe protein was amplified by polymerase chain reaction and cloned into e. coli vector pqe30 ua. the recombinant plasmid designated as pq pe was transformed into e. coli m15 cells and induced with iptg revealed the high level expression of 11.9 kda his-fusion protein as estimate ...201628330270
responses of bovine innate immunity to mycobacterium avium subsp. paratuberculosis infection revealed by changes in gene expression and levels of microrna.paratuberculosis in cattle is a chronic granulomatous gastroenteritis caused by mycobacterium avium subsp. paratubercolosis (map) which is endemic worldwide. in dairy herds, it is responsible for huge economic losses. however, current diagnostic methods do not detect subclinical infection making control of the disease difficult. the identification of map infected animals during the sub-clinical phase of infection would play a key role in preventing the dissemination of the pathogen and in reduci ...201627760169
seroprevalence of infectious diseases in saiga antelope (saiga tatarica tatarica) in kazakhstan 2012-2014.286 serum samples were collected from three sub-populations of saiga in kazakhstan (betpakdala, ustyurt and volga-ural) between 2012 and 2014, and were tested for the presence of antibodies to brucella spp., bluetongue virus, peste des petits ruminants (ppr) virus, akabane virus, schmallenberg virus, chlamydophila, toxoplasma, mycobacterium avium subspecies paratuberculosis and coxiella burnetii (q fever). seropositives to coxiella burnetii of saiga were detected and the adjusted seroprevalence ...201627094147
exposure to viral and bacterial pathogens among soay sheep (ovis aries) of the st kilda archipelago.we assessed evidence of exposure to viruses and bacteria in an unmanaged and long-isolated population of soay sheep (ovis aries) inhabiting hirta, in the st kilda archipelago, 65 km west of benbecula in the outer hebrides of scotland. the sheep harbour many metazoan and protozoan parasites but their exposure to viral and bacterial pathogens is unknown. we tested for herpes viral dna in leucocytes and found that 21 of 42 tested sheep were infected with ovine herpesvirus 2 (ohv-2). we also tested ...201626829883
the role of il-10 in mycobacterium avium subsp. paratuberculosis infection.mycobacterium avium subsp. paratuberculosis (map) is an intracellular pathogen and is the causative agent of johne's disease of domestic and wild ruminants. johne's disease is characterized by chronic granulomatous enteritis leading to substantial economic losses to the livestock sector across the world. map persistently survives in phagocytic cells, most commonly in macrophages by disrupting its early antibacterial activity. map triggers several signaling pathways after attachment to pathogen r ...201627905994
a mycobacterium avium subsp. paratuberculosis predicted serine protease is associated with acid stress and intraphagosomal survival.the ability to maintain intra-cellular ph is crucial for bacteria and other microbes to survive in diverse environments, particularly those that undergo fluctuations in ph. mechanisms of acid resistance remain poorly understood in mycobacteria. although, studies investigating acid stress in m. tuberculosis are gaining traction, few center on mycobacterium avium subsp. paratuberculosis (map), the etiological agent of chronic enteritis in ruminants. we identified a map acid stress response network ...201627597934
persistence of bacterial indicators and zoonotic pathogens in contaminated cattle wastes.manure can provide a favourable environment for pathogens' survival. de-contamination may be conducted by extended storage, until field conditions are suitable for application to land as source of agricultural nutrients.201627206734
paratuberculosis (johne's disease) in cattle and other susceptible species.paratuberculosis (johne's disease) is a widespread and costly disease. this consensus statement will summarize recommendations regarding diagnosis, control, and treatment of johne's disease in cattle and other species. each section of recommendations is followed by a statement that subjectively characterizes the strength of the supporting evidence. the role played by mycobacterium avium subsp. paratuberculosis (map) in the pathogenesis has been a matter of controversy for many years. this statem ...201623106497
assessing the impact of tailored biosecurity advice on farmer behaviour and pathogen presence in beef herds in england and wales.the term 'biosecurity' encompasses many measures farmers can take to reduce the risk of pathogen incursion or spread. as the best strategy will vary between settings, veterinarians play an important role in assessing risk and providing advice, but effectiveness requires farmer acceptance and implementation. the aim of this study was to assess the effectiveness of specifically-tailored biosecurity advice packages in reducing endemic pathogen presence on uk beef suckler farms. one hundred and sixt ...201627931934
molecular epidemiology of mycobacterium avium subsp. paratuberculosis on dairy farms.mycobacterium avium subspecies paratuberculosis (map) is the etiological agent of severe chronic intestinal inflammatory disease in ruminants, termed johne's disease, and can infect many other animal species, including humans. map has a long incubation period prior to manifestation of clinical signs including diarrhea, weight loss, and loss of production. map has a high prevalence in dairy herds and results in considerable adverse impacts on animal health and productivity throughout the world. r ...201626526547
prevalence of mycobacterium avium subsp. paratuberculosis fecal shedding in alpacas presented to veterinary hospitals in the united states.the prevalence of johne's disease in alpacas in the united states is unknown. the limits of polymerase chain reaction (pcr) detection of mycobacterium avium subsp. paratuberculosis (map) in alpaca feces have not been determined.201623782278
a new compartmental model of mycobacterium avium subsp. paratuberculosis infection dynamics in cattle.models of mycobacterium avium subsp. paratuberculosis (map), a chronic infectious agent of cattle, are used to identify effective control programs. however, new biological findings show that adult infections occur and that infected animals can be separated into 2 paths: animals that will become high-shedding and, eventually, experience clinical disease (high-path); and animals that will shed only small quantities of map and will remain subclinical (low-path). longitudinal data analysis found tha ...201526520176
whole-blood gene-expression profiles of cows infected with mycobacterium avium subsp. paratuberculosis reveal changes in immune response and lipid metabolism.mycobacterium avium subsp. paratuberculosis (map) is the causative agent of johne's disease, a chronic debilitating disease affecting ruminants worldwide. in the present study, we aimed to determine the major gene networks and pathways underlying the immune response to map infection using whole-blood cells, as well as provide the potential transcriptional markers for identifying the status of map infection. we analyzed the transcriptional profiles of whole-blood cells of cattle identified and gr ...201525248984
crohn's disease-specific anti-cuzd1 pancreatic antibodies are absent in ruminants with paratuberculosis.pancreatic autoantibodies (pabs) specifically recognizing gp2 and/or cuzd1 are present in more than 35% of patients with crohn's disease (crd). we have recently provided evidence of the presence of gp2-specific pabs in ruminants with paratuberculosis (ptb), a mycobacterium avium paratuberculosis (map)-induced disease resembling crd.201525575461
sensitivity and specificity of repeated test results from a commercial milk enzyme-linked immunosorbent assay for detection of mycobacterium avium subspecies paratuberculosis in dairy cattle.to investigate the sensitivity and specificity of results of initial and repeated milk elisas (at 6- or 12-month intervals) to detect cows that were shedding mycobacterium avium subsp paratuberculosis (ie, were infectious) and to evaluate factors influencing the probability that the results of a repeated milk elisa would be positive for an infectious cow if the results of the initial milk elisa were negative.201525554941
envelope protein complexes of mycobacterium avium subsp. paratuberculosis and their antigenicity.mycobacterium avium subsp. paratuberculosis (map) is the causative agent of johne's disease, a chronic enteric disease of ruminant animals. in the present study, blue native page electrophoresis and 2d sds-page were used to separate map envelope protein complexes, followed by mass spectrometry (ms) to identify individual proteins within the complexes. identity of individual proteins within complexes was further confirmed by ms upon excision of spots from 2d sds-page gels. among the seven putativ ...201525500374
cellular composition of granulomatous lesions in gut-associated lymphoid tissues of goats during the first year after experimental infection with mycobacterium avium subsp. paratuberculosis.mycobacterium avium subsp. paratuberculosis (map) causes lesions in naturally and experimentally infected ruminants which greatly differ in severity, cellular composition and number of mycobacteria. morphologically distinct lesions are already found during the clinically inapparent phase of infection. the complex local host response and number of map were characterized at the initial sites of lesions, organized gut-associated lymphoid tissue, in experimentally infected goats. tissues were collec ...201525466387
sequential development of lesions 3, 6, 9, and 12 months after experimental infection of goat kids with mycobacterium avium subsp paratuberculosis.the development of lesions after infection with mycobacterium avium subsp paratuberculosis (map) was examined in an experimental infection model. goat kids were orally inoculated 10 times with 10 mg bacterial wet mass of map (total dose 2.6 × 10(8) colony-forming units). six to 7 inoculated goats and 3 controls were autopsied 3, 6, 9, and 12 months postinoculation (mpi), lesions were documented, and samples were collected for histology, immunohistochemistry (ihc), and bacterial culture. twenty-f ...201524829286
environmental risk factors in the incidence of johne's disease.this review addresses the survival and persistence of mycobacterium avium subsp. paratuberculosis (map), the causative pathogen of johne's disease (jd), once it has left its ruminant host. jd has significant economic impact on dairy, beef and sheep industries and is difficult to control due to the long-term sub-clinical nature of the infection, intermittent or persistent map shedding during and after this period, inadequate test effectiveness, and the potential for map to exist for extended peri ...201524670062
mycobacterium avium subsp. paratuberculosis sheep strains isolated from cyprus sheep and goats.paratuberculosis, caused by mycobacterium avium subsp. paratuberculosis (map), is a chronic incurable infection of intestinal tract of animals. molecular characterization of map isolates classifies them into two major groups, 'cattle' or type ii and 'sheep' or type i/iii with a different phenotype, epidemiology, virulence and pathogenesis. the aim of this study was to examine 192 map elisa-positive sheep and goats from cyprus using faecal culture and genotype map isolates using is1311 pcr and re ...201523683358
associations between mycobacterium paratuberculosis sero-status, milk quality parameters, and reproduction in dairy cows.the objective of this study was to investigate the influence of mycobocterium avium subsp. paratuberculosis (map) sero-status of dairy cows on different milk production variables and reproductive traits. the study was carried out on 40 herds from the region of galicia (north-west spain). these herds were randomly selected from a larger group that had taken part in a voluntary paratuberculosis control program since 2005, which involves regular serum sampling of every adult animal to run antibody- ...201526591382
[production and evaluation of a purified protein derivative from an argentine strain of mycobacterium avium subsp. paratuberculosis].purified protein derivatives (ppds) are non-defined antigens prepared from mycobacteria cultures. they are usually employed to evaluate the specific cellular immune response both in animals and humans. bovine and avian ppds are usually employed as antigens in mycobacterial infections such as tuberculosis and paratuberculosis. nevertheless, ppd from mycobacterium avium subsp. paratuberculosis, (ppdj) is neither commonly used nor frequently available. however, ppd from mycobacterium avium subsp. a ...201523102462
crohn's disease, the mycobacterium paratuberculosis and the genetic bond: an unexpected trio. 201525771330
crohn's disease therapy with dietzia: the end of anti-inflammatory drugs. 201525689526
serologic screening for 13 infectious agents in roe deer (capreolus capreolus) in flanders.in order to investigate the role of roe deer in the maintenance and transmission of infectious animal and human diseases in flanders, we conducted a serologic screening in 12 hunting areas.201526609692
lamp technology: rapid identification of brucella and mycobacterium avium subsp. paratuberculosis.in this study, we developed new sets of primers to detect brucella spp. and m. avium subsp. paratuberculosis (map) through isothermal amplification. we selected a previously well-characterized target gene, bscp31, specific for brucella spp. and is900 for map. the limits of detection using the loop-mediated isothermal amplification (lamp) protocols described herein were similar to those of conventional pcr targeting the same sequences. hydroxynaphtol blue and sybr green(tm) allowed direct naked-e ...201526273282
in vivo volatile organic compound signatures of mycobacterium avium subsp. paratuberculosis.mycobacterium avium ssp. paratuberculosis (map) is the causative agent of a chronic enteric disease of ruminants. available diagnostic tests are complex and slow. in vitro, volatile organic compound (voc) patterns emitted from map cultures mirrored bacterial growth and enabled distinction of different strains. this study was intended to determine vocs in vivo in the controlled setting of an animal model. vocs were pre-concentrated from breath and feces of 42 goats (16 controls and 26 map-inocula ...201525915653
characterization of a caprine model for the subclinical initial phase of mycobacterium avium subsp. paratuberculosis infection.paratuberculosis caused by mycobacterium avium subsp. paratuberculosis (map) is difficult to control due to a long phase of clinically non-apparent (latent) infection for which sensitive diagnostics are lacking. a defined animal model for this phase of the infection can help to investigate host-map interactions in apparently healthy animals and identify surrogate markers for disease progress and might also serve as challenge model for vaccines. to establish such a model in goats, different age a ...201525889716
efficient, validated method for detection of mycobacterial growth in liquid culture media by use of bead beating, magnetic-particle-based nucleic acid isolation, and quantitative pcr.pathogenic mycobacteria are difficult to culture, requiring specialized media and a long incubation time, and have complex and exceedingly robust cell walls. mycobacterium avium subsp. paratuberculosis (map), the causative agent of johne's disease, a chronic wasting disease of ruminants, is a typical example. culture of map from the feces and intestinal tissues is a commonly used test for confirmation of infection. liquid medium offers greater sensitivity than solid medium for detection of map; ...201525609725
granuloma coinfection with mycobacterium bovis, mycobacterium avium subsp. paratuberculosis, and corynebacterium pseudotuberculosis in five hunted red deer (cervus elaphus) in portugal.we report granulomatous lymphadenitis in red deer (cervus elaphus) in portugal caused by coinfection with corynebacterium pseudotuberculosis, mycobacterium bovis, and mycobacterium avium subsp. paratuberculosis, as demonstrated by molecular methods.201525932668
analysis of the bovine monocyte-derived macrophage response to mycobacterium avium subspecies paratuberculosis infection using rna-seq.johne's disease, caused by infection with mycobacterium avium subsp. paratuberculosis, (map), is a chronic intestinal disease of ruminants with serious economic consequences for cattle production in the united states and elsewhere. during infection, map bacilli are phagocytosed and subvert host macrophage processes, resulting in subclinical infections that can lead to immunopathology and dissemination of disease. analysis of the host macrophage transcriptome during infection can therefore shed l ...201525699042
the identification of circulating mirna in bovine serum and their potential as novel biomarkers of early mycobacterium avium subsp paratuberculosis infection.mycobacterium avium subspecies paratuberculosis (map) is the aetiological agent of johne's disease (jd), a chronic enteritis in ruminants that causes substantial economic loses to agriculture worldwide. current diagnostic assays are hampered by low sensitivity and specificity that seriously complicate disease control; a new generation of diagnostic and prognostic assays are therefore urgently needed. circulating micrornas (mirnas) have been shown to have significant potential as novel biomarkers ...201526218736
analysis of biobanked serum from a mycobacterium avium subsp paratuberculosis bovine infection model confirms the remarkable stability of circulating mirna profiles and defines a bovine serum mirna repertoire.johne's disease (jd) is a chronic enteritis of ruminants caused by mycobacterium avium subspecies paratuberculosis (map). current disease control strategies are hampered by the lack of sensitive and specific diagnostic modalities. therefore, novel diagnostic and prognostic tools are needed, and circulating micrornas (mirnas) may hold potential in this area. the aims of this study were twofold: (i) to address the stability of mirna in bovine sera from biobanked samples, and (ii) to assess the pot ...201526675426
impact of t300a variant of atg16l1 on antibacterial response, risk of culture positive infections, and clinical course of crohn's disease.autophagy-related 16-like 1 (atg16l1) deficiency leads to impaired cellular autophagy and bacterial degradation as well as an altered cytokine production. the single-nucleotide polymorphism rs2241880 (t300a) is associated with an increased risk for crohn's disease (cd). atg16l1 polymorphisms could therefore have an impact on the risk of infectious complications and disease course in cd. we examined the impact of the t300a genotype on the antibacterial response toward a panel of pathogenic bacter ...201526673830
prevalence of mycobacterium avium subsp. paratuberculosis and escherichia coli in blood samples from patients with inflammatory bowel disease.mycobacterium avium subsp. paratuberculosis (map) and adherent-invasive escherichia coli (aiec) have been implicated as primary triggers in crohn's disease (cd). in this study, we evaluated the prevalence of map and e. coli (ec) dna in peripheral blood from 202 inflammatory bowel disease (ibd) patients at various disease periods and compared against 24 cirrhotic patients with ascites (cir) (non-ibd controls) and 29 healthy controls (hc). map dna was detected by is900-specific nested pcr, ec dna ...201525994082
pathogenesis of crohn's disease: bug or no bug.the possibility of an infectious origin in inflammatory bowel disease (ibd) has been postulated since the first description of crohn's disease (cd). many observations implicate bacteria as a trigger for the development of cd: lesions occur in regions with higher bacterial concentrations; aphthous ulcers occur in peyer's patches; inflammation resolves when the fecal stream is diverted and is reactivated following reinfusion of bowel contents; severity of the disease is correlated with bacterial d ...201525685606
human interferon regulatory factor 5 homologous epitopes of epstein-barr virus and mycobacterium avium subsp. paratuberculosis induce a specific humoral and cellular immune response in multiple sclerosis patients.a large number of reports indicate the association of epstein-barr virus (ebv), and mycobacterium avium subsp. paratuberculosis (map) with multiple sclerosis (ms).201525392335
mycobacterium paratuberculosis as a cause of crohn's disease.crohn's disease is a chronic inflammatory bowel disease of unknown cause, affecting approximately 1.4 million north american people. due to the similarities between crohn's disease and johne's disease, a chronic enteritis in ruminant animals caused by mycobacterium avium paratuberculosis (map) infection, map has long been considered to be a potential cause of crohn's disease. map is an obligate intracellular pathogen that cannot replicate outside of animal hosts. map is widespread in dairy cattl ...201526474349
lack of humoral response against helicobacter pylori peptides homologous to human znt8 in hashimoto's thyroiditis patients.the helicobacter pylori (hp) reinfection rate seems to be higher in developing countries than in developed ones. an increased seroprevalence of hp has also been reported in patients with type 1 diabetes (t1d) and hashimoto's thyroiditis (ht). mycobacterium avium subsp. paratuberculosis (map) has been linked to both t1d and ht. quite a few lines of evidence indicate that autoantibodies against several epitopes belonging to human zinc transporter 8 (znt8) cross-recognize the homologous map3865c ep ...201526142673
mycobacterium avium subspecies hominissuis in crohn's disease: a case report.we have cultured mycobacteria avium subspecies hominissuis (mah) from the blood of a patient with crohn's disease. the patient is a 21 year-old-female with a diagnosis of crohn's disease for two years. she had been treated with corticosteroids and humira for six months. a blood specimen was cultured in a specialized medium, and there was visible bacterial growth present in the liquid culture medium after eight weeks. pcr analysis of the bacterial growth and subsequent direct sequencing of the pc ...201526510454
species-specific engagement of human nucleotide oligomerization domain 2 (nod)2 and toll-like receptor (tlr) signalling upon intracellular bacterial infection: role of crohn's associated nod2 gene variants.recognition of bacterial peptidoglycan-derived muramyl-dipeptide (mdp) by nucleotide oligomerization domain 2 (nod2) induces crucial innate immune responses. most bacteria carry the n-acetylated form of mdp (a-mdp) in their cell membranes, whereas n-glycolyl mdp (g-mdp) is typical for mycobacteria. experimental murine studies have reported g-mdp to have a greater nod2-stimulating capacity than a-mdp. as nod2 polymorphisms are associated with crohn's disease (cd), a link has been suggested betwee ...201525335775
the treatment of infectious disease with a medical device: results of a clinical trial of ultraviolet blood irradiation (uvbi) in patients with hepatitis c infection.prior to the advent of therapies with sustained virological response rates of 94%, this study was conducted for the us food and drug administration (fda) to assess the safety and efficacy of ultraviolet blood irradiation (uvbi) for the treatment of hepatitis c virus (hcv) infection.201526092299
on deaf ears, mycobacterium avium paratuberculosis in pathogenesis crohn's and other diseases.the historic suggestion that mycobacterium avium subsp. paratuberculosis (map) might be a zoonotic pathogen was based on the apparent similarity of lesions in the intestine of patients with crohn's disease (cd) with those present in cattle infected with map, the etiological agent of johne's disease. reluctance to fully explore this possibility has been attributed to the difficulty in demonstrating the presence of map in tissues from patients with cd. advances in technology have resolved this pro ...201526730151
iron acquisition in mycobacterium avium subsp. paratuberculosis.mycobacterium avium subsp. paratuberculosis is a host-adapted pathogen that evolved from the environmental bacterium m. avium subsp. hominissuis through gene loss and gene acquisition. growth of m. avium subsp. paratuberculosis in the laboratory is enhanced by supplementation of the media with the iron-binding siderophore mycobactin j. here we examined the production of mycobactins by related organisms and searched for an alternative iron uptake system in m. avium subsp. paratuberculosis. throug ...201526712939
zap-70, ctla-4 and proximal t cell receptor signaling in cows infected with mycobacterium avium subsp. paratuberculosis.paratuberculosis is a chronic intestinal disease of ruminant animals caused by mycobacterium avium subsp. paratuberculosis (map). a hallmark of paratuberculosis is a transition from a cell-mediated th1 type response to a humoral th2 response with the progression of disease from a subclinical to clinical state. the objective of this study was to investigate the expression of two crucial molecules in t cell function, zap-70 (zeta-chain-associated protein of 70 kda) and ctla-4 (cytotoxic t-lymphocy ...201526163934
application of multiple laboratory tests for mycobacterium avium ssp. paratuberculosis detection in crohn's disease patient specimens.the difficulties involved in detecting and enumerating mycobacterium avium subsp. paratuberculosis (map) as a pathogen potentially involved in crohn's disease (cd) are well known. this study aimed to improve this situation through the application of multiple laboratory diagnostic tests to detect and isolate this bacterium from different specimens collected from cd-patients and non-cd subjects as controls. a total of 120 samples (terminal ileum and colon biopsies, blood and stool) were obtained f ...201526147146
functional analysis of 3'utr polymorphisms in the caprine slc11a1 gene and its association with the mycobacterium avium subsp. paratuberculosis infection.the study aimed to investigate whether the genetic polymorphisms in the 3'utr of the caprine slc11a1 gene are functional, and to assess the role of map as a regulatory parameter in gene expression. to this goal we constructed plasmids expressing the luciferase reporter gene in transient transfections of a mouse (balb/c) macrophage cell line (raw264.7), incorporating those polymorphisms that our previous work indicated as more prominent in terms of slc11a1 expression and responsiveness to map inf ...201526117376
bovine immunoinhibitory receptors contribute to suppression of mycobacterium avium subsp. paratuberculosis-specific t-cell responses.johne's disease (paratuberculosis) is a chronic enteritis in cattle that is caused by intracellular infection with mycobacterium avium subsp. paratuberculosis. this infection is characterized by the functional exhaustion of t-cell responses to m. avium subsp. paratuberculosis antigens during late subclinical and clinical stages, presumably facilitating the persistence of this bacterium and the formation of clinical lesions. however, the mechanisms underlying t-cell exhaustion in johne's disease ...201526483406
macrophage polarization in cattle experimentally exposed to mycobacterium avium subsp. paratuberculosis.mycobacterium avium subspecies paratuberculosis (map), the causative agent of johne's disease (jd) in cattle, has significant impacts on the livestock industry and has been implicated in the etiology of crohn's disease. macrophages play a key role in jd pathogenesis, which is driven by the manipulation of host immune mechanisms by map. a change in the macrophage microenvironment due to pathogenic or host-derived stimuli can lead to classical (m1) or alternative (m2) polarization of macrophages. ...201526454271
host transcriptional profiles and immunopathologic response following mycobacterium avium subsp. paratuberculosis infection in mice.paratuberculosis or johne's disease is a chronic granulomatous enteropathy in ruminants caused by mycobacterium avium subsp. paratuberculosis (map) infection. in the present study, we examined the host response to map infection in spleens of mice in order to investigate the host immunopathology accompanying host-pathogen interaction. transcriptional profiles of the map-infected mice at 3 and 6 weeks p.i. showed severe histopathological changes, whereas those at 12 weeks p.i. displayed reduced le ...201526439498
application of cattle slurry containing mycobacterium avium subsp. paratuberculosis (map) to grassland soil and its effect on the relationship between map and free-living amoeba.slurry from dairy farms is commonly used to fertilize crops and pastures. this mixture of manure, urine and water can harbor multiple microbial pathogens among which mycobacterium avium subsp. paratuberculosis (map) is a major concern. persistence of map in soil and infection of soil acanthamoeba was evaluated by culture, real-time is900 pcr, and by staining of amoeba with acid-fast and vital stains comparing soils irrigated with map-spiked or control dairy farm slurry. map dna was detected in s ...201525448447
molecular analysis and miru-vntr typing of mycobacterium avium subsp. paratuberculosis strains from various sources.mycobacterium avium subsp. paratuberculosis (map) is the causative agent of johne's disease. genotypic discrimination of map isolates is pivotal to epidemiological studies requisite for revealing infection sources and disease transmission. this study was undertaken to determine the genetic diversity of map strains from diverse sources.201525410632
identification of immunoreactive proteins of mycobacterium avium subsp. paratuberculosis.mycobacterium avium subsp. paratuberculosis (map) is the cause of a chronic enteritis of ruminants (bovine paratuberculosis (ptb)--johne's disease) that is associated with enormous worldwide economic losses for the animal production. diagnosis is based on observation of clinical signs, the detection of antibodies in milk or serum, or evaluation of bacterial culture from feces. the limit of these methods is that they are not able to detect the disease in the subclinical stage and are applicable o ...201525404104
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