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La Paratuberculosis Bovina (PTB) o Enfermedad de Johne, es una infección del tracto gastrointestinal causada por Mycobacterium avium, subespecie paratuberculosis (Map), que se caracteriza por generar enteritis granulomatosa crónica y linfadenitis en rumiantes. La preocupación más relevante en relación con la importancia de la PTB es su posible vínculo con la Enfermedad de Crohn (EC) en humanos, sin embargo, esta asociación aún está bajo investigación. Se determinó la seroprevalencia de PTB en el municipio de Sogamoso (Boyacá), donde se recolectaron 604 muestras de sangre, cuyo suero fue procesado mediante la técnica de ELISA indirecta con el kit comercial PARACHEK® 2 KIT (Prionics, Suiza). La seroprevalencia fue de 10,9% (66/604), donde el grupo etario de 2 a 3 años y la raza Jersey fueron los de mayor seroprevalencia. Se encontró asociación estadística significativa (p≤0,05) entre la edad de los individuos evaluados y el suministro de concentrado. La seroprevalencia encontrada indica que se está produciendo una transmisión activa de la enfermedad y que las medidas de control disponibles no están siendo llevadas a cabo o no son lo suficientemente efectivas.

Abstract Current Johne’s disease control programs primarily focus on decreasing transmission of Mycobacterium avium subsp. paratuberculosis (MAP) from infectious adult cows to susceptible calves. However, potential transmission between calves is largely overlooked. The objective was to determine the extent of MAP infection in calves contact-exposed to infectious penmates. Thirty-two newborn Holstein–Friesian calves were grouped into 7 experimental groups of 4, consisting of 2 inoculated (IN) calves, and 2 contact-exposed (CE) calves, and 1 control pen with 4 non-exposed calves. Calves were group housed for 3 months, with fecal samples were collected 3 times per week, blood and environmental samples weekly, and tissue samples at the end of the trial. The IN calves exited the trial after 3 months of group housing, whereas CE calves were individually housed for an additional 3 months before euthanasia. Control calves were group-housed for the entire trial. All CE and IN calves had MAP-positive fecal samples during the period of group housing; however, fecal shedding had ceased at time of individual housing. All IN calves had MAP-positive tissue samples at necropsy, and 7 (50%) of the CE had positive tissue samples. None of the calves had a humoral immune response, whereas INF-γ responses were detected in all IN calves and 5 (36%) CE calves. In conclusion, new MAP infections occurred due to exposure of infectious penmates to contact calves. Therefore, calf-to-calf transmission is a potential route of uncontrolled transmission on cattle farms.

Paratuberculosis (PTB) is caused by Mycobacterium avium subsp. paratuberculosis (MAP) and is one of the most widespread and economically important diseases in cattle. After birth, calves are raised with natural breast feeding without separation from their mothers in most Korean native cattle (Hanwoo breed) farms. Vertical transmission of PTB has been reported, but the exact PTB infection route has not been revealed in Hanwoo farms. Calves of MAP seropositive dams were tested for MAP presence and MAP antibodies in feces and tissues. MAP was detected in calf tissues by using polymerase chain reaction. Expressions of genes reported to be prognostic biomarkers of MAP infection changed in both calves and cows (p ＜ 0.05). Expression of two genes (HGF and SERPINE1) were significantly decreased in MAP-infected cattle and their offspring (p ＜ 0.01). The results suggest that biomarker gene expression profiles can be useful in detecting early stage MAP infection. Based on the results, complete eradication of MAP may be possible if accurate diagnostic methods to detect infected calves are added to the current PTB eradication strategy, which, because infected individuals are likely to develop into fecal MAP shedders at any time, includes isolation of new born calves and feeding sterilized colostrum. [ABSTRACT FROM AUTHOR]

Background: Inflammatory bowel disease (IBD) is described as a relapsing condition with high morbidity and uncertain complex pathogenesis. The association of Mycobacterium avium ssp. paratuberculosis (MAP) with Crohn's disease (CD) in human has been debated for decades, however there is no confirmed data to verify such relations in Iran. The aim of this study was to investigate risk factors and a possible role of MAP in Iranian patients with CD. Methods: Anti-MAP antibodies were detected in serum of IBD patients and subjects without IBD (nIBD) through ELISA; MAP DNA and viable MAP cells were identified in patients' biopsies through nested PCR and direct culture methods, respectively. Principal component analysis (PCA) was used to investigate the risk factors in relation to IBD and MAP infection. Results: Positivity for IS900 PCR was detected in 64% (n = 18) of CD, 33% (n = 10) of ulcerative colitis (UC) and 9.7% (n = 6) of nIBD samples. Live MAP cells were isolated from biopsies of 2 CD patients only. Among 28 patients with CD, 46% (n = 13) and 39% (n = 11) were positive for antibodies against MAP3865c133-141 and MAP3865c125-133 peptides, respectively, whereas much lower seroreactivity was detected in UC subjects accounting for 3% (n = 1) for MAP3865c133-141 and 16.7% (n = 5) for MAP3865c125-133. A high immune reactivity to MAP epitopes among CD patients was positively correlated with consumption of fast food meals and IBD familiarity. For both CD and UC, breastfeeding period and consumption of fruit/vegetables presented negative correlation with the presence of anti- MAP antibodies. Conclusions: This study provided evidences that high prevalence of MAP DNA and anti-MAP antibodies in CD patients is significantly associated with the development of CD. Despite the role of several factors contributing to IBD, the presence of MAP DNA and anti-MAP antibodies in Iranian CD patients highlights a possible transmission of MAP from animal-derived products to humans. [ABSTRACT FROM AUTHOR]

Fibronectin attachment protein (FAP) also known as ModD gene of 987 bp size was identified in the Mycobacterium avium subsp. paratuberculosis (MAP) genome. This gene was successfully amplified by PCR using specific primers and MAP genomic DNA template, in the presence of 10% DMSO and glycerol. The PCR product was cloned in pGEMT easy cloning vector and subsequently subcloned into pET-33b expression vector and sequenced. MAP ModD gene showed sequence identity ranging from 64 to 94% at nucleotide level and 47– 87% at amino acid level, by WU-BLAST program with FAPs of other Mycobacteria species. MAP ModD gene showed 97% sequence homology with M. avium subsp avium ModD by Megalign tool of DNAStar programme. This strong identity was attributed to the more close genetic association of these two species. MAP ModD protein is mainly composed of beta sheets and is highly antigenic. Positive pETModD recombinant clone was induced with IPTG for expression of the ModD protein. The MAP ModD protein was purified under native condition using nickel affinity column. The expressed recombinant ModD (52 kDa) was confirmed by Western blot with nickel- HRPO conjugate and rabbit hyperimmune serum raised against recombinant ModD. [ABSTRACT FROM AUTHOR]

مایکوباکتریوم آویوم زیر گونه پاراتوبرکلوزیس(MAP) عامل سببی بیماری یون یا Paratuberculosis از جمله بیماری‌های عفونی مزمن مهم از نظر اقتصادی است که گریبانگیر صنعت گاوداری و نشخوارکنندگان می‌باشد. این بیماری به صورت آنتروکولیت گرانولوماتوز، لنفادنیت و تورم عروق لنفاوی موضعی ظاهر می‌شود که علامت شاخص این بیماری کاهش وزن پیشرونده بیمار است. با توجه به اهمیت تشخیص این بیماری در این بررسی از دو روش کشت و واکنش زنجیره‌ای پلیمراز برای شناسایی این میکروارگانیسم استفاده شد. در این بررسی 100 نمونه شیر از گاوهای به‌ظاهر سالم و 100 نمونه شیر از گاوهای مشکوک به ابتلاء به بیماری یون مربوط به تعدادی از گاوداری‌های منطقه شهرستان سراب، آذربایجان شرقی، ایران انجام گرفت. پس از جمع‌آوری نمونه‌ها٬ آزمایش مستقیم میکروسکوپی با رنگ‌آمیزی اختصاصی ذیل نلسون صورت گرفت. سپس کشت باکتریایی روی محیط اختصاصی تهیه شده و در نهایت با روش واکنش زنجیره‌ای پلیمراز با استفاده از پرایمرهای اختصاصی شناسایی مایکوباکتریوم اویوم پاراتوبرکلوزیس بررسی شد. براساس نتایج آزمایش مستقیم، کشت و واکنش زنجیره‌ای پلیمراز بر روی شیر100 راس گاو به ظاهر سالم، به ترتیب 8، 9 و 12 مورد مثبت و در بررسی این تست‌ها بر روی شیر 100 راس گاو مشکوک به ابتلاء به بیماری یون به ترتیب 15، 40 و44 مورد مثبت شناسایی شد. نتایج حاصل از این بررسی نشان داد آزمایش کشت و واکنش زنجیره‌ای پلیمراز در شناسایی عامل بیماری یون بسیار حائز اهمیت هستند. لذا با توجه به فراوانی این بیماری در منطقه مورد مطالعه می‌توان از هرکدام از روش‌های یاد شده در شناسایی این میکروارگانیسم استفاده کرد.

Mycobacterium avium subsp. paratuberculosis (MAP) is a well-established etiological agent of Johne’s disease in animals. In humans, similar clinical condition, first described by Crohn as regional ileitis in 1932, now known as Crohn’s diseases (CD), has also been associated with this mycobacterial species. However, there are two schools of thoughts, one favoring MAP as its etiological agent while the second considers it as an immune-inflammatory condition triggered by an external factor. Onset of CD requires a series of events including predisposition of certain inherited genetic traits, associated environmental stimuli, and immune-inflammatory response. A combination of these factors probably leads to this disease. Recently, some human genes have also been identified which regulate ability to respond appropriately to the external factors. Added to these factors are concerns about the selection of clinical specimens and poor adherence to laboratory quality controls. The literature is full of contradictory findings, but there a lack of uniformity in the materials and methods used by many of these researchers. In this review, we provide our perspective under above circumstances and give our point of view which may open a platform for debate regarding the MAP as the etiological agent of human CD.

This paper describes the clinical, pathological, and microbiologic aspects of paratuberculosis (Johne's disease) in a dairy Gyr herd in the State of Paraíba, northeastern Brazil. An eight years old cow with chronic unresponsive diarrhea was clinically examined and euthanized for pathological evaluation. Fecal samples from all 160 animals over 12 months of age from the herd were collected for isolation of Mycobacterium avium subsp. paratuberculosis. Clinically, the index case cow was severely dehydrated, cachectic, with profuse mucous diarrhea. The main post-mortem findings were emaciation and thickened intestinal wall. Microscopically, the intestinal lamina propria and submucosa were infiltrated by macrophages, epithelioid cells, and Langhans giant cells with numerous alcohol-acid resistant bacilli in the cytoplasm. Two fecal samples displayed growth in slants of Herrold's egg-yolk agar supplemented with mycobactin J, 150 days after incubation. No growth was noticed in slants without mycobactin J. Microscopic examination of the isolated microorganisms stained by Ziehl-Neelsen revealed considerable amounts of alcohol-acid resistant bacilli, morphologically compatible with Mycobacterium spp. Based on the clinical signs, gross and histological lesions, growth time, bacterial morphology in Ziehl-Neelsen staining, and dependence of mycobactin J, the first diagnosis of paratuberculosis in Zebu cattle was made.Objetivou-se descrever os aspectos clínicos, anátomo-histopatológicos e microbiológicos da paratuberculose em um rebanho Gir leiteiro no Estado da Paraíba. Uma vaca de oito anos que apresentava diarréia persistente, refratária a tratamento foi necropsiada para estudo anátamo-histopatológico. Também foram coletadas amostras de fezes de todos os 160 animais do plantel, com idade superior a 12 meses, para tentativa de isolamento de Mycobacterium avium subsp. paratuberculosis. Ao exame clínico, o animal caso índice apresentou caquexia, diarréia profusa e desidratação grave. À necropsia, o animal apresentou-se emaciado e, ao exame detalhado do trato digestivo, foi observado espessamento da parede e superfície mucosa do íleo e intestino grosso. À microscopia, verificou-se intensa infiltração de macrófagos espumosos associado a raras células epiteliódes e gigantes do tipo Langerhans na lâmina própria e submucosa. À coloração de Ziehl-Neelsen foram observadas miríade de bacilos álcool-ácido resistentes no citoplasma destas células. Houve crescimento de colônias bacterianas em duas das 160 amostras de fezes após 150 dias de incubação em tubos com meio Herrold's egg-yolk suplementados com micobactina J e ausência de crescimento nos tubos com mesmo meio, mas sem suplementação. Os microrganismos isolados foram corados pelo Ziehl-Neelsen observando-se presença de grande quantidade de bacilos álcool-ácido resistente, com morfologia compatível ao gênero Mycobacterium. Baseado na história clínica, achados anátomo-histopatológicos e histoquímicos (Ziehl-Neelsen), e microbiológicos, firmou-se o primeiro diagnóstico de paratuberculose em Zebu na Paraíba.

Johne’s disease or paratuberculosis is a chronic granulomatous enteritis in ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). The disease is responsible for significant economic losses in dairy industry worldwide. Microbial culture as a golden standard test for detection of MAP in faecal specimens requires 6-16 weeks to complete, whereas accurate and rapid identification of cattle shedding MAP in their feces is essential for successful control of the disease in herds. In the present study, direct microbial diagnosis by Ziehl-Neelsen acid-fast staining, microbial culture on Herrolds’ egg yolk media and two IS900 direct PCR assays were carried out on 100 fecal specimens of apparently healthy cattle collected from dairy herds of Tabriz with a history of Johne’s disease. The number of positive specimens identified by the direct microbial diagnosis, microbial culture and PCR with F90/F91 and FP25/FP26 primes were 7 (7%), 14 (14%), 15(15%) and 25(25%) respectively. These results indicated that PCR detected more positive cases therefore it can be employed for rapid and accurate diagnosis of cattle shedding MAP it their feces and the type of primer used has a significant role in the sensitivity of this test. Direct microbial diagnosis by Ziehl-Neelsen acid-fast staining identified 7 (7%) specimens, two IS900 direct PCR assays identified 15 (15%) and 25 (25%) specimens, respectively, and microbial culture identified 7 (7%) specimens as positive. Collectively, these data indicate that PCR detection of MAP was more sensitive than direct microbial diagnosis by Ziehl-Neelsen acid-fast staining or faecal culture, especially if appropriate primers were used.

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 paratuberculosis has been considered as an alternative strategy to control the disease when combined with management interventions. Understanding host-pathogen interactions is extremely important to development of vaccines. It has long been known that Th1-mediated cellular immune responses are play a crucial role in protection against MAP infection. However, recent studies suggested that innate immune responses are more closely related to protective effects than adaptive immunity. Based on this understanding, several attempts have been made to develop vaccines against paratuberculosis. A variety of ideas for designing novel vaccines have emerged, and the tests of the efficacy of these vaccines are conducted constantly. However, no effective vaccines are commercially available. In this study, studies of the development of vaccines for MAP were reviewed and summarized. [ABSTRACT FROM AUTHOR]

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 or high fiber diet during challenge. The composition of microbiota of the cecal content and the sacculus rotundus was studied in 20 New Zealand white female rabbits. The extracted DNA was subjected to paired-end Illumina sequencing of the V3-V4 hypervariable region of the 16S rRNA gene for microbiota analysis. Microbial richness (Chao1) in the cecal content was significantly increased by MAP infection in regular diet rabbits (p = 0.0043) and marginally increased (p = 0.0503) in the high fiber group. Analysis of beta-diversity showed that MAP infection produces deeper changes in the microbiota of sacculus rotundus than in the cecal content. A lower abundance of Proteobacteria in the cecal content of infected animals fed the high fiber diet and also lower abundance of Bacteroidetes in the sacculus rotundus of infected animals fed the regular diet were observed. Based on OPLS-DA analysis, we observed that some bacteria repeatedly appear to be positively associated with infection in different samples under different diets (families Dehalobacteriaceae, Coriobacteriaceae, and Mogibacteriaceae; genus Anaerofustis). The same phenomenon was observed with some of the bacteria negatively associated with MAP infection (genera Anaerostipes and Coprobacillus). However, other groups of bacteria (Enterobacteriaceae family and ML615J-28 order) were positively associated with infection in some circumstances and negatively associated with infection in others. Data demonstrate that MAP infection and diet changes do interact and result in shifts in the microbiota of the cecal content and sacculus rotundus of rabbits. [ABSTRACT FROM AUTHOR]

Mycobacterium avium subsp. paratuberculosis (MAP) is an etiological agent of chronic inflammation of the intestine among ruminants and humans. Currently, there are no effective vaccines and sensitive diagnostic tests available for its control and detection. For this, it is of paramount importance to identify the MAP antigens, which may be immunologically recognized by the host immune system. To address this challenge, we performed identification of the immunogenic epitopes in the MAP outer membrane proteins (OMPs). We have previously identified 57 MAP proteins as OMPs [Rana A, Rub A, Akhter Y. 2014. Molecular BioSystems, 10:2329-2337] and have evaluated them for the epitope selection and analysis employing a computational approach. Thirty-five MAP OMPs are reported with nine-mer peptides showing high binding affinity to major histocompatibility complex (MHC) class I molecules and 28 MAP OMPs with 15-mer peptides of high binding affinity for MHC class II molecules. The presence of MHC binding epitopes indicates the potential cell-mediated immune response inducing capacity of these MAP OMPs in infected host. To further investigate the humoral response inducing properties of OMPs of MAP, we report potential B cell epitopes based on the sequences of peptide antigens and their molecular structures. We also report 10 proteins having epitopes for both B and T cells representing potential candidates which may invoke both humoral and cellular immune responses in the host. These findings will greatly accelerate and expedite the formulation of effective and cost-efficient vaccines and diagnostic tests against MAP infection. Copyright © 2015 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Background: Rabbits are susceptible to infection by different species of the genus Mycobacterium. Particularly, development of specific lesions and isolation of Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis, both subspecies of the M. avium complex, has been reported in wildlife conditions. Although, rabbit meat production worldwide is 200 million tons per year, microbiological data on this source of meat is lacking and more specifically reports of mycobacterial presence in industrially reared rabbit for human consumption have not been published. To this end, we sought mycobacteria by microbiological and histopathological methods paying special attention to Mycobacterium avium subsp. paratuberculosis in rabbits from commercial rabbitries from the North East of Spain. Results: M. avium subsp. paratuberculosis was not detected either by culture or PCR. However, Mycobacterium avium subsp. avium was detected in 15.15 % (10/66) and Mycobacterium avium subsp. hominissuis was detected in 1.51 % (1/66) of gut associated lymphoid tissue of sampled animals by PCR, whereas caecal contents were negative. 9 % (6/66) of the animals presented gross lesions suggestive of lymphoid activation, 6 % (4/66) presented granulomatous lesions and 3 % (2/66) contained acid fast bacilli. Mycobacterial isolation from samples was not achieved, although colonies of Thermoactinomycetes sp. were identified by 16s rRNA sequencing in 6 % (4/66) of sampled animals. Conclusions: Apparently healthy farmed rabbits that go to slaughter may carry M. avium subspecies in gut associated lymphoid tissue. [ABSTRACT FROM AUTHOR]

Background: paratuberculosis (PTB) is clinically characterized by diarrhea and progressive weight loss in ruminants, and causes economic losses due to decreased milk production, premature animal disposal, and increased veterinary treatment costs. The presence of PTB has been increasing in milk producing bovine farms in Mexico, yet epidemiology data from specific farms remains scarce. Objective: to determine the proportion of cows positive for Mycobacterium avium subsp. paratuberculosis (Map) among culled cows. Methods: in the present study, 40 culled cows from 25 farms located at the complejo agropecuario industrial de Tizayuca (CAITSA) in Hidalgo (Mexico) were evaluated for PTB presence by histopathological and microbiological studies. Tissue and rectal scraping samples were taken from all cows, and studied for presence of histopathology PTB-compatible lesions. Samples were processed for Map isolation on selective agar culture media; typical Map colonies were confirmed by polymerase chain reaction (PCR). Results: potential PTB infection was found in 25% (10/40) of the animals evaluated by histopathology and acid-fast bacilli (AFB) staining, but only 7.5% (3/40) were positive for type 2 Map (C cattle strain) by PCR. Conclusions: the prevalence of PTB in cattle farms at CAITSA was confirmed, which may be valuable epidemiological information in Mexico. [ABSTRACT FROM AUTHOR]

In last decades, the prevalence and importance of Johne's disease is increased, and consequently different programs are designed for its control. Johne's disease is considered the most important disease of cattle industry regarding the economic lost. In addition they are published several data, which implicate the Mycobacterium avium subsp. paratuberculosis as a pathogen bacteria of Chrohn's disease in humans. The chronic pattern of disease, complexity of its path biology, limitations knowledge's on pathogenesis, poor sensitivity and specificity of currently available laboratory diagnostic methods, lack of successful vaccines, make Johne's disease control a significant challenge for veterinary services and scientific community as well. This paper aim to highlight the current achievements on path-biology, diagnosis, biosecurity, in order to translate them in practical action for Johne's disease control. [ABSTRACT FROM AUTHOR]

Mycobacterium avium subsp. paratuberculosis (MAP) has a high degree of resistance to chemical and physical procedures frequently used for the elimination of other bacteria. Recently, a method for the determination of viability by exposure of MAP to propidium monoazide (PMA) and subsequent real time quantitative PCR (qPCR) was established and found to be comparable with culture. The aim of this study was to apply the PMA qPCR method to determine the impact of increasing concentration or time and repeated cycles of the application of selected disinfectants on MAP viability. Different MAP isolates responded to the same type of stress in different ways. The laboratory strain CAPM 6381 had the highest tolerance, while the 8819 low-passage field isolate was the most sensitive. Ultraviolet exposure caused only a partial reduction in MAP viability; all MAP isolates were relatively resistant to chlorine. Only the application of peracetic acid led to the total elimination of MAP. Repeated application of the treatments resulted in more significant decreases in MAP viability compared to single increases in the concentration or time of exposure to the disinfectant. [ABSTRACT FROM AUTHOR]

Paratuberculosis, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is predominantly a disease of ruminants. The microorganism is shed principally in the feces of affected animals, even without clinical signs. Young animals, up to the age of 6 months are at the highest risk of infection by oral exposure to contaminated feces. There is no treatment and vaccination is not permitted in Israel. The standard method of reducing farm infection rates is by improving management hygiene and culling shedding cows, especially super-shedders (more than 104 copies of the hspX gene). Consequently the prompt identification of animals shedding MAP is of cardinal importance in eradicating the infection. The aim of this preliminary study was to devise a fecal sampling protocol that would maximize the likelihood of detecting MAP shedding cows and assessing the influence of parturition induced stress on shedding. Sixteen cows and 15 heifers, raised on a commercial dairy farm of about 350 lactating cows, with 10% milk MAP ELISA positive cows were included in the survey. Rectal feces, sampled for 5-7 consecutive days, before and after calving, were examined by quantitative PCR (qPCR). Results indicate that the microorganism was shed by positive animals regularly and quantitative variations were minimal. Consequently one sample is a good indication of the animal's shedding status. MAP shedding was found not be influenced by parturition associated stress. [ABSTRACT FROM AUTHOR]

In this study the suitability of different solid media was investigated for the isolation of Mycobacterium avium subsp. paratuberculosis (Map) in order to identify the optimum single or combination of media to permit the isolation of all strain types from small ruminants. A subset of these Map strains was then further characterized by molecular typing methods to assess the genetic diversity of Map strains in the study area (Northern Greece). Map strains were isolated from tissues and faeces of infected goats (n = 52) and sheep (n = 8) and were analysed for polymorphisms in IS1311 to classify the strain type as Type C or S. The study found that M7H11 supplemented with mycobactin j, OADC and new born calf serum (M7H11+Mj) is the best single choice of medium for the primary isolation of Map of both Type C and S from small ruminants. The combination of M7H11+Mj and Herrolds egg yolk medium supplemented with mycobactin j and sodium pyruvate allowed the detection of all Map isolates in this study. Nineteen Map isolates were characterised by pulsed-field gel electrophoresis and the isolates demonstrated significant genetic diversity. Twelve different SnaBI and 16 distinct SpeI profiles were detected of which 25 have not been described previously and are new profiles. The combination of both enzyme profiles gave 13 different multiplex profiles. Ten different multiplex profiles were detected in goats and three in sheep. One ovine isolate gave the same multiplex profile as a caprine isolate and two different profiles were found within a single goat herd. [ABSTRACT FROM AUTHOR]

Paratuberculosis caused by Mycobacterium avium subsp. paratuberculosis (MAP) is a chronic infectious disease affecting domestic and wild ruminants. Antigens currently used for the diagnosis of paratuberculosis are whole-cell derived crude preparations. The identification of MAP-specific antigens for the specific and early diagnosis of this infection is strongly needed. This study assessed the ability of the MAP-specific synthetic lipopeptide antigen Para-LP-01 to invoke specific serum antibody (Ab) and cellmediated immune (CMI) responses in sheep experimentally exposed to MAP S strain. Responses were compared to those elicited by the crude whole-cell derived MAP 316v antigen (316v). Para-LP-01 induced a significant serum Ab response in MAP-infected sheep in comparison with unexposed or uninfected sheep, but failed to induce detectable CMI responses including production of IFN-γ, IL-10 and lymphoproliferation, unlike 316v which invoked both CMI and serum Ab responses in MAP-exposed sheep. Para-LP- 01 is a suitable antigen for serodiagnosis of MAP-infection in sheep. The differential induction of humoral and CMI responses by lipid based antigens could enhance current understanding of the role played by cell-wall associated lipid antigens in the pathogenesis of MAP-infection. [ABSTRACT FROM AUTHOR]

The article presents a study that provides new data on mycobacterial infection among wild boars. It investigated the presence of mycobacterium avium subspecies paratuberculosis (MAP) in blood and feces collected from 222 hunter-killed wild boars in Korea from December 2010 to March 2011. It examined the blood and fecal samples by culture, polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay. It identified 2 major MAP genotypes infecting domestic species in Korea.

The article explains the role of lymphatic fluid in screening Mycobacterium avium subspecies paratumberculosis (MAP) in cattle. The study was performed on 58 cows, where lymph fluid was extracted from their udders to identify traces of MAP through nested polymerase chain reaction. Results revealed that MAP was identified in 43.1 percent of lymph samples, where it was discovered in 66.7 percent cows with clinical Johne's disease.

The article discusses a study of the first detection of Mycobacterium avium subspecies paratuberculosis (Map) in skeletal muscle and blood from mixed-age ewes with and without Johne's disease in a New Zealand farm. The ewe flock at the farm did not receive Johne's vaccination. Results showed 71% of ewes with Johne's disease have positive cultures. Also noted is the presence of low numbers of Map in the blood of the infected ewes.

Paratuberculosis is an enteric disease caused by Mycobactaium avium Subsp. paratuberculosis (Map) in domestic and Wild ruminants. Johne's disease causes huge production losses and has high Impact on animal industry. Due to the lack of Simple and effiCient diagnostic tests, the control programmes are hampered espeCially for subclinically infected animals. Various diagnostic tools are available like fecal culture techniques, culturing, radiOisotyping, PCR based methods, serological approaches (Cell mediated assay and humoral based) and most new area are recombinant antigens based detetion which can open a novel approach for early detection of the disease. [ABSTRACT FROM AUTHOR]

The association between Mycobacterium avium subsp. paratuberculosis (MAP) and Crohn's disease (CD) is supported by several studies reporting the detection or isolation of MAP from human tissues, but a direct association is still debatable. Objective. To evaluate the survival of MAP in human intestinal cells and to measure the presence of antibodies against two mycobacterial proteins necessary for the survival of the bacterium in the sera of CD patients. Material and methods. Human-derived intestinal cells were infected with three isolates of MAP and the survival of the microorganism was determined. The presence of antibodies against protein tyrosine phosphatase A (PtpA) and protein kinase G (two proteins secreted within the host in the early stages of the invasion) in the sera of CD patients was evaluated. Sera of 20 CD patients and 20 controls were collected and the presence of the antibodies was assayed using enzyme-linked immunosorbent assay (ELISA). Secretion of the PtpA in vivo was visualized by immunostaining. Results. MAP survived in intestinal cells, and immunostaining of PtpA showed that the protein was secreted within these cells. Wilcoxon rank sum test revealed that CD patient sera had significantly higher titer of antibodies specific for both of these antigens compared to controls. ELISA results for either protein were not statistically different between men and women. Conclusions. The presence of specific antibodies against mycobacterial proteins essential for establishing an infection in the host suggests that MAP can potentially be active in CD patients, and a serological test can be developed for early detection of MAP in CD patients. [ABSTRACT FROM AUTHOR]

Background: Mycobacterium avium subsp. paratuberculosis (MAP) persistently infects intestines and mesenteric lymph nodes leading to a prolonged subclinical disease. The MAP genome sequence was published in 2005, yet its transcriptional organization in natural infection is unknown. While prior research analyzed regulated gene sets utilizing defined, in vitro stress related or advanced surgical methods with various animal species, we investigated the intracellular lifestyle of MAP in the intestines and lymph nodes to understand the MAP pathways that function to govern this persistence. Results: Our transcriptional analysis shows that 21%, 8% and 3% of the entire MAP genome was represented either inside tissues, macrophages or both, respectively. Transcripts belonging to latency and cell envelope biogenesis were upregulated in the intestinal tissues whereas those belonging to intracellular trafficking and secretion were upregulated inside the macrophages. Transcriptomes of natural infection and in vitro macrophage infection shared genes involved in transcription and inorganic ion transport and metabolism. MAP specific genes within large sequence polymorphisms of ancestral M. avium complex were downregulated exclusively in natural infection. Conclusions: We have unveiled common and unique MAP pathways associated with persistence, cell wall biogenesis and virulence in naturally infected cow intestines, lymph nodes and in vitro infected macrophages. This dichotomy also suggests that in vitro macrophage models may be insufficient in providing accurate information on the events that transpire during natural infection. This is the first report to examine the primary transcriptome of MAP at the local infection site (i.e. intestinal tissue). Regulatory pathways that govern the lifecycle of MAP appear to be specified by tissue and cell type. While tissues show a "shut-down" of major MAP metabolic genes, infected macrophages upregulate several MAP specific genes along with a putative pathogenicity island responsible for iron acquisition. Many of these regulatory pathways rely on the advanced interplay of host and pathogen and in order to decipher their message, an interactome must be established using a systems biology approach. Identified MAP pathways place current research into direct alignment in meeting the future challenge of creating a MAP-host interactome. [ABSTRACT FROM AUTHOR]

The article discusses research on the diagnosis of the Mycobacterium avium subspecies of bovine colostrum called paratuberculosis. It references a study by Patrick Pithua and colleagues, published in the 2010 issue of the "Journal of Veterinary Diagnostic Investigation." The researchers examined the use of a nested polymerase chain reaction (PCR) in detecting the subspecies. They found that the PCR method is effective in identifying the presence of paratuberculosis in colostrum.

Validity of Johne's disease programs and control protocols that rely on established cut points [e.g., specified sample-to-positive (S/P) ratios] for ELISA serological tests depends on interpreted results that are not susceptible to variable test accuracy. It was hypothesized that seasonal variability exists in serological response to Mycobacterium avium subsp, paratuberculosis (MAP) infection. Further, a reciprocal response may occur, resulting in greater risk of fecal shedding in subclinically infected animals. A testing regimen was invoked that included multiple testing of individual adult cows during the 4 seasons. Serum was collected on a cyclic, monthly basis from 3 randomly selected cohorts of dairy cows, and fecal samples were collected from the 20% of cows with the greatest ELISA test S/P ratios. Staggered, quarterly sampling was continued for 1 yr, and at the conclusion, serum was analyzed en masse. The ELISA outcome values (i.e., S/P ratio) were treated both as categorical and continuous variables. The potential lagged effects of temperature-related seasonality on S/P ratio, as well as the potential for a change in test result caused by temperature were assessed. Results for fecal culture were analyzed on a categorical scale and compared with the ELISA results to explore the possibility of reciprocal fecal shedding. No significant seasonal effects on either S/P ratios or the proportion of cows seropositive to MAP were observed. Furthermore, no evidence was found linking temperature-related seasonality to a reciprocal increase in the risk of fecal culture positivity for MAP. [ABSTRACT FROM AUTHOR]

The article presents a case study of two Eurasian otters with Mycobacterium avium paratuberculosis (MAP), who were killed after collision with a vehicle in Castelo Branco, Portugal. As mentioned, postmortem examination revealed no significant gross alterations in organs but showed that both animals had diffuse lymphadenitis with macrophage infiltration. Further, it is stated that acid-fast organisms were isolated from gastrointestinal tissue samples and were identified as MAP.