SEARCH · Search PubMed
Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
Restriction endonuclease analysis and DNA hybridization revealed five ovine strains of Mycobacterium paratuberculosis from South Africa had identical DNA patterns to an ovine strain from Canada. Genetically this strain type has features in common with the two major groups of M. paratuberculosis.
In this study the effect of supplementation of drinking water with mycobactin J or lactoferrin on the multiplication of Mycobacterium paratuberculosis in gnotobiotic mice was investigated. The results indicated that neither mycobactin J nor lactoferrin, at the doses used, appeared to have any effect on fecal shedding or tissue burdens of M. paratuberculosis.
Data from an epidemiological study in Ontario, involving 304 dairy herds, were used to identify associations between selected production indices and lipoarabinomannan antigen serological test results for paratuberculosis (LAM-ELISA). Analyses were conducted at both the herd and individual cow levels of organization. After analytically controlling for management and cow factors in the respective regression models, positive serological paratuberculosis status (as defined by the LAM-ELISA test), was associated with higher milk somatic cell counts at both the herd average (p less than 0.01), and individual cow levels of organization (p less than 0.0001). In contrast, LAM-ELISA test results were consistently not associated with calving intervals in either the herd average or individual cow level analyses. Associations between LAM-ELISA results and milk production were inconsistent. No associations were found at the herd level of organization, and LAM-ELISA results were not associated with a change in breed class average (BCA) for milk, between the previous and the most recent lactations of individual cattle. However, at the individual cow level, LAM-ELISA results were positively associated with higher milk production as measured by the current BCA (p less than 0.05), and individual cow average kg of milk produced per year of life since two years of age (p less than 0.0001).
Mycobacterium paratuberculosis was diagnosed in 2 llamas by bacteriologic culturing and DNA probe testing of fecal and intestinal specimens. Diarrhea and weight loss were observed in both animals. Lesions in the gastrointestinal tracts of the 2 llamas were similar to pathologic changes reported in cattle infected with M paratuberculosis.
A random sample of Wisconsin dairy herds, stratified by herd size, were tested for paratuberculosis by use of an absorbed ELISA procedure. The ELISA was optimized for overall accuracy by means of receiver operating characteristic curve analysis, and had a sensitivity and specificity of 50.9 and 94.9%, respectively. Herd prevalence was analyzed for correlation with responses to a management practices questionnaire completed by the herd owners. One hundred and fifty-eight herds and 4,990 cattle were tested. Of these, 50% of herds and 7.29% of cattle had positive test results. Calculation of true prevalence from the apparent prevalence indicated that 4.79% of cattle and 34% of the Wisconsin dairy herds tested had serologic evidence of paratuberculosis. Among the 54 herds classified as positive on the basis of true prevalence estimation, the mean number of test positive cattle was 20.3%. The geographic distribution of herds with positive results was not uniform. More infected herds were found in the southern and western districts of Wisconsin than in the eastern district. The west-central district had a larger number of infected herds than did other districts. By use of chi 2 analysis, the only management factor found to be significantly associated with herd prevalence was housing of calves after weaning (P = 0.03). Specifically, in herds with higher prevalence, calves were separated after weaning into calf barns and hutches rather than into pens in the cow barn more often than in herds with lower prevalence. This factor was also considered significant by use of logistic regression analysis.(ABSTRACT TRUNCATED AT 250 WORDS)
OBJECTIVE: To compare milk with serum ELISA for detection of antibodies to Mycobacterium paratuberculosis. DESIGN: Epidemiologic survey. ANIMALS: 821 Missouri dairy cattle of 12 herds that were brucellosis certified and Dairy Herd Improvement Association members. PROCEDURE: Milk and serum samples obtained concurrently from Missouri dairy herds were tested by use of a standard ELISA procedure. Concurrent collection of milk and serum samples controlled for interactions such as colostral antibodies and the effect of time. On the basis of milk and serum ELISA values, disease prevalence and correlation between milk and serum test results were determined. RESULTS: Correlation measures on individual animals indicated low correlation of milk and serum ELISA values. McNemar's chi 2 of 7.6 was significant (P = 0.05). Analysis correlation was low (kappa = 0.08), as was regression analysis (R2 = 0.02). CLINICAL IMPLICATIONS: Milk ELISA for the detection of exposure to M paratuberculosis lacked correlation with serum ELISA. Further evaluation to determine sensitivity and specificity of milk ELISA will augment the usefulness of milk ELISA as a herd screening test.
Mycobacterial culture was performed on colostrum, milk, and feces from 126 clinically normal cows of a single herd with high prevalence of Mycobacterium paratuberculosis infection. Thirty-six (28.6%) cows were determined to be shedding the organism in the feces. Of the 36 fecal Culture positive cows, M paratuberculosis was isolated from the colostrum of 8 (22.2%) and from the milk of 3 (8.3%). Cows that were heavy fecal shedders were more likely to shed the organism in the colostrum than were light fecal shedders.
Developments in the diagnosis and treatment of paratuberculosis is constrained by the lack of an experimental animal model. To investigate this problem conventional chicks immunodepressed by a Cyclophosphamide injection and concurrent inoculation of Infectious Bursal Disease Virus (IBDV) were infected with Mycobacterium paratuberculosis and kept for 4 months. The immunodepressed chicks eliminated mycobacteria with their faces from the first month until the third month and developed typical intestinal lesions of mycobacterial infection characterized by aggregation of macrophages with monocytes and lymphocytes. Diarrhoea was absent. The number of lymphocytes decreased by about 80%. The serological tests carried out with Complement Fixation test were negative. For the positive bacteriology and typical granulomatous lesions, the conventionally reared chicks proved to be a useful laboratory model for reproduction of Johne's disease.
OBJECTIVE: To investigate the relation between lesions and cellular immune response-based tests in sheep with naturally acquired paratuberculosis and to evaluate efficacy of an interferon-gamma (IFN-gamma) assay and the intradermal skin test. ANIMALS: 110 and 88 adult sheep were used to evaluate the IFN-gamma assay and skin test, respectively. PROCEDURE: Before euthanasia, heparinized blood was collected and incubated with avian purified protein derivative (PPD). Interferon-y production was assessed, using a commercial enzyme immunoassay. Skin testing in the caudal fold also was performed, using avian PPD. Sensitivity and specificity of these techniques were evaluated, using histopathologic findings as reference. RESULTS: Of the sheep examined, 61 (55.4%) had lesions that were classified in 5 categories, according to location, extension, cellular types, and presence of acid-fast bacilli. Regarding cellular immunity test results, 55.4% of sheep with and 6.1 % without pathologic changes had positive results of the IFN-gamma assay. Skin test sensitivity (55.6%) and specificity (100%) were slightly greater. CONCLUSION: Close association between pathologic findings and cellular immune response was apparent. Most sheep with focal (tuberculoid) and some with diffuse lesions (borderline tuberculoid), all with scant or no acid-fast bacilli in the intestine, had positive results of the IFN-gamma assay and skin test. Most sheep with diffuse lesions and high numbers of bacilli (borderline lepromatous) were test negative. CLINICAL RELEVANCE: Cellular immune response-based and serologic tests are useful tools for seeking evidence of paratuberculosis. Each technique detects different types of immune-reacting sheep, and their simultaneous use may detect most infected animals.
Johne's disease is a chronic enteritis of cattle and other ruminant species that is of worldwide economic importance. The cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) have been associated with granuloma formation and wasting in other disease syndromes. The potential role of these cytokines in the development and progression of Johne's disease has not been investigated. Using reverse transcriptase polymerase chain reaction (RT-PCR) and specific bovine oligonucleotide cytokine primers and probes for bovine TNF-alpha and IL-6, we examined the ex vivo expression of mRNA for these inflammatory cytokines in whole blood from healthy cattle. Cytokine mRNA levels increased after a brief incubation of bovine whole blood with Mycobacterium paratuberculosis or its lipoarabinomannan (LAM). Muramyl dipeptide (MDP) and Escherichia coli LPS also stimulated TNF-alpha and IL-6 mRNA expression. Several strains of M. paratuberculosis were tested and found to have similar abilities to stimulate TNF-alpha and IL-6 mRNA expression. Several strains of the closely related Mycobacterium avium, and the unrelated saprophyte, Mycobacterium phlei, had somewhat less ability to stimulate TNF-alpha and IL-6 mRNA expression.
IgG antibodies against Mycobacterium paratuberculosis protoplasmic antigen were looked for by an enzyme immunosorbent assay in patients with Crohn's disease, Ulcerative colitis, active pulmonary tuberculosis, past pulmonary tuberculosis, and in healthy controls. Serum reactivity for these antibodies was not correlated to PPD skin test positivity without history of mycobacterial disease. A cutoff based on the mean absorbance value of a pool of healthy blood donors was chosen. Positive values were found in 12/24 (50%) patients with active pulmonary tuberculosis and 2/10 (20%) skin-test-positive subjects with past pulmonary tuberculosis as expected, because antigenic sharing is common among different mycobacteria. The control group of blood donors showed borderline positivities in 3/149 cases (2.01%). Positive values were found in 4/108 (3.70%) patients with Crohn's disease and 2/40 (5%) with ulcerative colitis. We conclude that our data do not support a causal relationship between the M. paratuberculosis and Crohn's disease, but occasional patients with inflammatory bowel diseases show unexpected positivities for these mycobacterial antibodies.
Recent data using improved cultural, molecular, and serological techniques have strengthened the association of Mycobacterium paratuberculosis with Crohn's disease, an inflammatory bowel disease (IBD) with unknown etiology. To provide more evidence of an etiological association, antibody reactivities of Crohn's disease patients were tested by immunoblotting against M. paratuberculosis-recombinant antigens. A clone containing a 1,402-bp insert and expressing a 36K-antigen (p36) was analyzed. No homology was found between the deduced amino acid sequence of p36 and any protein sequences compiled in the GenBank indicating that p36 is a novel mycobacterial protein. The reactivity of 199 serum samples was tested against the p36 by immunoblotting technique. Sera from 77 of 89 (86.5%) Crohn's disease patients and 16 of 18 (89%) sera from patients with tuberculosis and leprosy reacted with p36 compared to 5 of 42 (12%) ulcerative colitis and non-IBD control sera (p < 0.0001). In addition, p36 reacted to all sera from 10 normal controls that were Bacillus Calmette-Guerin (BCG)-immunized and only to 10% of 40 normal controls that were not BCG-immunized. The fact that sera from Crohn's disease patients reacted to p36 with the same high frequency as the sera from patients that were exposed to mycobacterial antigens further supports the hypothesis of the mycobacterial etiology in Crohn's disease.
A novel method for the lysis and subsequent fractionation of bacterial constituents from Mycobacterium paratuberculosis strain 19698 (M. paratuberculosis) and Escherichia coli strain DH5 alpha utilizing the technique of matrix solid-phase dispersion (MSPD) is described. Bacteria were blended with octadecylsilyl (C18) derivatized silica to obtain cellular lysis. The blended material was used to prepare a column which was sequentially eluted with solvents of increasing polarity. Fractionation of cellular components was confirmed by analysis of the solvent extracts. The possible applicability of the MSPD technique as a general method for the lysis and fractionation of bacterial components is proposed.
Serum IgG, IgM and IgA antibody response in 20 cattle naturally infected with Mycobacterium paratuberculosis and in 15 non-infected cattle were measured by enzyme-linked immunosorbent assay. A strong IgG response was detected in 16 (80%) of the infected animals. Diagnostic levels of IgM were detectable in all of the infected animals as well as in 8 (53%) of the non-infected animals. Animals with paratuberculosis had a very weak specific serum IgA response and this appears to be of little value in detection of infection in these animals.
Two groups of C57 and C3H mice of 5 weeks of age were infected via the intraperitoneal route with 2.0 mg (wet weight) of Mycobacterium paratuberculosis. These were maintained with a similar number of segregated and non-inoculated mice of the same strains under specially controlled conditions of low, medium and high iron intake. Mice were killed and bled at 7 months post-infection and assessments of haematological parameters and the degree of mycobacterial granulomatous involvement of abdominal and other tissues were made. In addition, the total mycobacterial numbers visible in macrophages in standardized histological sections of liver, spleen and bone marrow in the presence or absence of stainable iron storage compounds were assessed using a double staining technique for iron and mycobacteria. Moderate to marked anaemia in both C57 and C3H mice on low iron intake, irrespective of infection, indicated that an effective low iron status was achieved in the animals by dietary manipulation. Medium and high iron intake groups exhibited normal haematological parameters. Iron storage compounds were readily visible in liver microgranulomas of mice on medium and high, but not on low iron intake. In liver, spleen and bone marrow samples, mycobacterial counts in iron-containing microgranulomas were significantly higher than in those without stainable iron. Increased frequencies of residual and progressive infection were associated with increased iron intake. The greater susceptibility of the C57 strain was evident from the significantly higher liver microgranuloma counts, higher mycobacterial numbers and greater progressive infection when compared with the C3H strain. These findings in mice strongly suggest that slow multiplication of M. paratuberculosis is enhanced in iron-replete compared with iron-deficient macrophages. This enhancement occurs despite the capacity of the less susceptible strain of mouse to limit the spread of the organism within the body.
The sonicate antigen (MPS) of a local strain (IVRI) of Mycobacterium paratuberculosis and a commercial lysate of Strain 18 were analysed using hyperimmune rabbit and calf antisera to MPS in crossed immunoelectrophoresis with intermediate gel (CIE-ig) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The rabbit antiserum was more potent than the calf antiserum and it precipitated 35 and 15 antigens, respectively, among MPS and lysate antigens. SDS-PAGE resolved 50 and 32 peptides among these antigens respectively, of which, 35 and 15 were precipitated by rabbit antiserum. A CIE-ig reference system, with 30 MPS antigens, was standardized and used to analyse antibody specificities among sera derived from animals experimentally and naturally infected with bovine paratuberculosis. Fourteen antigens of MPS were found to be reactive with these sera and among these, Antigens 2 and 5 were found to be serodominant; sonicate antigens of M. bovis BCG and M. avium did not contain these antigens. Both were high molecular weight (greater than 60 kDa) antigens which may be of serodiagnostic value.
DNA of 90 mycobactin-dependent strains of Mycobacterium paratuberculosis, isolated in 9 countries, was digested with restriction endonuclease PstI and hybridized with a DNA fragment containing insertion sequence IS900. Bovine strains (n = 73) were isolated from 61 animals in 17 herds, ovine strains (n = 15) from 13 animals in 3 herds and the set was completed by 1 caprine and 1 human (Linda) strain. Three types, tentatively designated A (n = 37), B (n = 51) and C (n = 2) were differentiated by restriction fragment length polymorphism (RFLP). Of the bovine strains 27, 45 and 1 were classified as belonging to types A, B and C, respectively; of the 15 ovine strains 10 and 5 belonged to types A and B, respectively; the caprine strain belonged to type C. The human strain Linda, isolated in the U.S.A., from a man with Crohn's disease, belonged to B type. A certain degree of type uniformity was observed among strains isolated within one herd in the course of several years, and the prevalence of a single type was also recorded within individual regions of the Czech Republic and Slovakia. Type A was identified in the course of 2 years in a sheep farm with frequent sales and purchases of animals, and type C was demonstrated in a goat kept in the same farm. Differences between RFLP types of the strains isolated from mesenteric lymph nodes and intestinal mucosa were found in one cow and one sheep. Selected strains of M. paratuberculosis RFLP type A (14 strains) and B (18 strains) were digested with restriction endonuclease BstEII. All the strains of A type were classified into C1 group and all the strains of B type into Cx group.
Four calves and four goat kids were inoculated perorally with a Norwegian goat-pathogenic strain of Mycobacterium paratuberculosis. None of the calves developed clinical disease, pathological lesions or humoral antibody response, but the organism was reisolated from the small intestine and mesenteric lymph nodes when the calves were slaughtered after 7, 12, 14 and 18 months, respectively. As one of the goats died of non-specific causes and one did not become infected, two remained as positive controls. One of these became subclinically and one clinically infected, but both showed distinct histopathological lesions at necropsy. Both were shown to be positive in the complement fixation test (CFT) and the enzyme-linked immunosorbent assay (ELISA) but only the clinically affected goat proved positive in the agar gel immunodiffusion (AGID) test. The AGID test was found to be of low diagnostic value, and the ELISA was as sensitive as the CFT in detecting infection at an early stage. However, when infection was finally established, the ELISA titres became far higher than the CFT titres. The results confirm previous experience in Norway, that the current Norwegian strain of M. paratuberculosis has little or no pathogenicity for cattle.