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PubMed1990

[Allergic and serologic diagnosis of paratuberculosis in cattle].

The use of allergenodiagnosis in cattle herds where paratuberculosis is to be eradicated depends on the allergenogenic properties of the causative agent of this disease. An evaluation of the reliability of an intradermal allergic test performed to diagnose paratuberculosis in cattle in the given area revealed that this method is important especially if the result of this test is explicitly negative. A cultivation proof of the causative agent of paratuberculosis is of greatest importance among bacteriological methods for infection diagnosis. The agreement of the bacteriological and serological examination which reached 80% in young cattle pointed out the high specificity of an indirect haemagglutination test. In cows, unlike calves and heifers, the specificity of diagnostic tests was considerably lower. It is recommended to use an elimination method mainly in young cattle: regular examinations of calves from their age of three months will help to form a group of heifers free of infection, observing the basic infection controlling measures and providing conventient diet.

Animals↗
PubMed1990

Preliminary observations on ovine paratuberculosis (Johne's disease) in Zambia.

In an imported flock of sheep from South Africa, an ewe became partially anorexic and gradually losing weight and conditions. The paratuberculosis (Johne's disease) was confirmed on clinical, faecal and histopathological examination. Cultural examination remained doubtful. Serological investigation of other sheep in the flock and at other farm reacted to antibodies of Mycobacterium paratuberculosis on complement fixation test. The study suggests that the disease is actively spreading. This is the first report of ovine paratuberculosis in the Republic of Zambia.

Animals↗
PubMed1988

Accidental self-inoculation with Mycobacterium paratuberculosis bacterin (Johne's bacterin) by veterinarians in Wisconsin.

We surveyed Wisconsin veterinarians to assess the frequency and severity of accidental self- and other human exposure to Mycobacterium paratuberculosis bacterin (Johne's bacterin). Of 199 veterinarians administering the bacterin to cattle, 22 reported one or more exposures, including 19 needle-stick exposures, 8 skin surface exposures, and 2 oral mucosa exposures. The mean incidence of needle sticks was 5.5/100 veterinarians/year of bacterin use or 1/1,000 doses administered. The mean total doses given in the needle-stick exposure group was 276 +/- 318 vs 80 +/- 268 in the group without needle-stick exposure, and the mean number of months administering the bacterin was 21.7 and 16.1, respectively; 63% of needle-stick exposures took place during the injection process. Five adverse reactions were reported, and each resulted from needle-stick exposure. The only systemic reaction followed an exposure to the original bacterin formulation of sonically ruptured M paratuberculosis in Freund incomplete adjuvant. The remaining reactions were to the current formulation of whole killed M paratuberculosis in mineral oil and ranged from a small nodule persisting for 4 to 6 months to painful inflammation of a finger persisting for 24 months. We anticipate an increase in incidence of these minimally debilitating injuries as the use and distribution of the bacterin expands. For hand wounds, we recommend conservative management. Surgical intervention should be considered if a granuloma persists and causes the patient functional difficulty.

Accidents, Occupational↗
PubMed1986

In vitro transformation of lymphocytes from blood and milk of cows with subclinical paratuberculosis.

Lymphocytes from blood or milk of 12 cows were evaluated in vitro for the lymphocyte's capability to proliferate in response to mitogens (phytohemagglutinin-A, concanavalin A, and pokeweed mitogen) and to an antigen prepared from Mycobacterium paratuberculosis (purified protein derivative, PPD-J). Responses of 4 control cows were compared with those of 4 cows subclinically infected with M paratuberculosis and with 4 apparently noninfected herdmates. Blood lymphocytes or milk lymphocytes from control cows had no detectable responses to PPD-J. Blood lymphocytes from infected cows had significant (P less than 0.05) responses to PPD-J, but milk lymphocytes from these cows did not. Conversely, milk lymphocytes from apparently noninfected herdmate cows had significant (P less than 0.05) responses to PPD-J, but blood lymphocytes from these cows did not. There were no significant differences in the responses of blood lymphocytes from control, noninfected, or infected cows to the mitogens. However, milk lymphocytes from infected cows had significantly (P less than 0.05) lower responses than did lymphocytes from the milk of control or noninfected cows to all mitogens. The decreased responsiveness of milk lymphocytes from cows subclinically infected with M paratuberculosis may indicate that immunocompetency of the mammary gland was altered.

Animals↗
PubMed1985

Characterization of Mycobacterium paratuberculosis of bovine, caprine, and ovine origin by gas-liquid chromatographic analysis of fatty acids in whole-cell extracts.

The cellular fatty acid composition of Mycobacterium paratuberculosis from bovine, ovine, and caprine origin (grown on Herrold's egg yolk medium) was determined by gas-liquid chromatography (GLC) and mass spectrometry and was compared with that of 19 other Mycobacterium spp. A species-specific fatty acid of M paratuberculosis was not demonstrable. Gas-liquid chromatographic analysis of cellular fatty acids was used to develop flow charts for the rapid identification of M paratuberculosis and other Mycobacterium spp. Two charts for distinguishing organisms were developed; one chart was based on GLC alone and the other based on GLC, growth rate, and chromogenicity. Seemingly, the GLC was reliable for detecting Mycobacterium spp rapidly.

Animals↗
PubMed1985

Lymphocyte blastogenesis, complement fixation, and fecal culture as diagnostic tests for paratuberculosis in North American wild ruminants and domestic sheep.

The efficacy of the lymphocyte blastogenesis and complement-fixation tests and fecal culture for detection of Mycobacterium paratuberculosis infection was assessed in bighorn sheep (Ovis canadensis), elk (Cervus elaphus nelsoni), mule deer (Odocoileus hemionus), white-tailed deer (O virginianus), bighorn X mouflon (O musimon) hybrid sheep, and domestic sheep. Spontaneously infected bighorns were tested at the time of capture; experimentally infected animals were tested monthly for 12 months or periodically for 36 months. Lymphocyte blastogenesis tests were conducted with peripheral blood mononuclear cells and protein antigens of M avium, M bovis, and M paratuberculosis. Best diagnostic results were obtained when M avium purified-protein derivative was used as antigen and 20% bovine fetal serum was incorporated in the culture medium; a positive test was defined as a stimulation index greater than or equal to 3.5. Test sensitivity and specificity, respectively, were 82% and 94% in hybrid sheep and were 72% and 100% in domestic sheep. Sensitivity and specificity, respectively, were 39% and 94% in elk and 53% and 92% in deer. When infection was determined in spontaneously infected bighorns by culture of M paratuberculosis and/or the presence of acid-fast bacilli in characteristic microscopic lesions, sensitivity was 75% and specificity was 87%. Fecal cultures and the complement-fixation tests seldom correctly identified infected animals.

Animals↗
PubMed1977

Application of the direct leukocyte-migration agarose test in cattle from a Mycobacterium paratuberculosis-infected herd.

Using johnin as the specific antigen, the direct leukocyte-migration agarose test was applied to 10 cows from a Mycobacterium paratuberculosis-infected herd. Nine of the cows did not have signs of paratuberculosis; 1 cow was clinically affected. The cows were slaughtered and, on the basis of histopathologic and cultural examinations of the intestines, were classified as infected or noninfected. Six of the healthy cows from which the bacterium was not isolated and the clinically affected cow reacted with migration indices lower than those of 3 healthy animals that were found to harbor the bacterium. It is suggested that the long incubation period of paratuberculosis could to some extent be due to immunologic tolerance.

Animals↗
PubMed1983

Eastern white-tailed deer as a reservoir of ruminant paratuberculosis.

Ten white-tailed deer were shot and killed at a Connecticut farm that had a 6-year history of bovine paratuberculosis, and organs from these animals were examined for evidence of paratuberculosis. Mycobacterium paratuberculosis was isolated from the cecal lymph node, terminal ileum, and ileocecal valve of 1 deer and from the cecal lymph node of another. Characteristic lesions and acid-fast bacilli were not observed. It was concluded that infected deer could serve as sources of infection for domestic stock.

Animals↗
PubMed1981

Bovine paratuberculosis III. An evaluation of a whole blood lymphocyte transformation test.

A whole blood lymphocyte transformation test was used to examine cattle with varying degrees of infection with Mycobacterium paratuberculosis. Minimally infected animals characteristically responded to johnin purified protein derivative in the lymphocyte transformation test but did not routinely react on serological and/or skin testing. Heavily infected animals showed considerable variation in their lymphocyte transformation responses to antigen and some of them were consistently unresponsive. Antigen induced lymphocyte transformation reactions were recorded in 7.6 to 41.5% of uninfected animals whose infection status was determined by bacteriology and/or histopathology. The number of positive reactions recorded in uninfected animals depended on the population, the larger percentage being found in a herd with a proven history of paratuberculosis. The potential of lymphocyte transformation as a diagnostic test for bovine paratuberculosis is discussed.

Animals↗
PubMed1978

Effect of dexamethasone on Mycobacterium paratuberculosis infection in hamsters.

Sixty hamsters were randomly divided into 4 equal groups: group 1--given only dexamethasone, group 2--given dexamethasone and exposed to Mycobacterium paratuberculosis, group 3--exposed to M paratuberculosis but not treated, and group 4--used as normal controls. Group 2 hamsters (treated with dexamethasone and exposed to the bacterium) had significantly higher bacterial counts in spleen and aggregated lymphatic nodules (Peyser's patches) than did those in group 3. None of the hamsters exposed to M paratuberculosis (groups 2 and 3) developed clinical signs of disease. All hamsters treated with the drug (groups 1 and 2) grew more slowly than did the hamsters not treated.

Animals↗
PubMed1977

Application of the direct leukocyte-migration agarose test in cattle naturally infected with Mycobacterium paratuberculosis.

Four cows with persistent diarrhea and complement-fixing antibodies to johnin were examined, using the leukocyte-migration agarose test (LMAT) with johnin as the specific antigen. The diagnosis of paratuberculosis (Johne's disease) was microscopically verified at necropsy. As control animals, 3 healthy cows and 3 cows with disease other than paratuberculosis were examined, using LMAT. The migration indices in the group of cows with paratuberculosis were significantly lower (less than 0.001) than those in the group of control cows. Furthermore, the migration zones of the unstimulated peripheral blood leukocytes in the former group were significantly smaller (less than 0.001) than those of the control group.

Animals↗
PubMed1985

Comparison of the pathogenicity of Mycobacterium paratuberculosis and Mycobacterium spp isolated from the wood pigeon (Columba palumbus-L).

The pathogenicity of 4 strains of mycobacteria isolated from wood pigeons and 2 strains of Mycobacterium paratuberculosis was compared in calves, mice and chickens. Three of the 4 wood pigeon strains and the 2 M. paratuberculosis strains produced clinical Johne's disease in calves. All 6 strains were pathogenic for mice and the 4 wood pigeon strains were pathogenic for chickens. The strains were not agglutinated by antisera to strains of the M. avium complex and all were mycobactin-dependent. It was concluded that the wood pigeon isolates may constitute a distinct group with the pathogenic capability of M. avium in chickens and M. paratuberculosis in calves.

Animals↗
PubMed1990

Effects of colonial morphology and tween 80 on antimicrobial susceptibility of Mycobacterium paratuberculosis.

Smooth (Sm) and rough (Rg) colonial types of Mycobacterium paratuberculosis ATCC 19698 and two clinical isolates were tested to examine their growth responses in medium containing antimicrobial agents. Susceptibility tests were done in Middlebrook 7H12B medium with and without Tween 80 and one of the following antimicrobial agents: streptomycin, isoniazid, rifampin, ethambutol, ciprofloxacin, and penicillin G. Growth responses in the presence of antimicrobial agents led to the following observations. (i) In the absence of Tween, Rg colony types were more resistant than Sm colony types; (ii) the addition of Tween 80 significantly increased the susceptibility of both Sm and Rg colony types; however, the increase was greater with the Sm colony types. These studies showed that the antimicrobial susceptibility of M. paratuberculosis was significantly affected when Tween 80 was present in either the primary culture medium or the drug susceptibility test medium. In the absence of the perturbing influence of Tween 80, M. paratuberculosis was resistant to the antimicrobial agents tested.

Anti-Bacterial Agents↗
PubMed2006

Mycobacterium avium subsp. paratuberculosis in lake catchments, in river water abstracted for domestic use, and in effluent from domestic sewage treatment works: diverse opportunities for environmental cycling and human exposure.

Mycobacterium avium subsp. paratuberculosis from infected animals enters surface waters and rivers in runoff from contaminated pastures. We studied the River Tywi in South Wales, United Kingdom, whose catchment comprises 1,100 km2 containing more than a million dairy and beef cattle and more than 1.3 million sheep. The River Tywi is abstracted for the domestic water supply. Between August 2002 and April 2003, 48 of 70 (68.8%) twice-weekly river water samples tested positive by IS900 PCR. In river water, the organisms were associated with a suspended solid which was depleted by the water treatment process. Disposal of contaminated slurry back onto the land established a cycle of environmental persistence. A concentrate from 100 liters of finished water tested negative, but 1 of 54 domestic cold water tanks tested positive, indicating the potential for these pathogens to access domestic outlets. In the separate English Lake District region, with hills up to 980 m, tests for M. avium subsp. paratuberculosis in the high hill lakes and sediments were usually negative, but streams and sediments became positive lower down the catchment. Sediments from 9 of 10 major lakes receiving inflow from these catchments were positive, with sediment cores indicating deposition over at least 40 to 50 years. Two of 12 monthly 1-liter samples of effluent and a single 100-liter sample from the Ambleside sewage treatment works were positive for M. avium subsp. paratuberculosis. Since Lake Ambleside discharges into Lake Windermere, which is available for domestic supply, there is a potential for these organisms to cycle within human populations.

Animals↗
PubMed1983

Iron-binding compounds of Mycobacterium avium, M. intracellulare, M. scrofulaceum, and mycobactin-dependent M. paratuberculosis and M. avium.

Fifty-three strains of M. avium and related species all produced one or more exochelins, the extracellular iron-binding compounds of the mycobacteria, when grown iron deficiently. Only those strains which could grow without the addition of mycobactin (i.e., mycobactin independent) produced mycobactin, the intracellular iron-binding compound of the mycobacteria. Exochelins varied from 20 to 2,000 micrograms per g of cell dry weight; mycobactins were between 1 and 10 mg per g of cell dry weight. M. paratuberculosis (13 strains) and 13 strains of M. avium, both species dependent upon mycobactin for growth, failed to produce spectrophotometrically detectable amounts of mycobactin (less than 0.2 microgram per g of cell dry weight), although mycobactin could be recognized in one strain of M. avium grown with an additional supply of salicylate and examined by a radiolabeling technique. On repeated subculture three of the mycobactin-dependent strains of M. avium, but none of those of M. paratuberculosis, lost their mycobactin dependence and on reexamination were found to produce their own mycobactin at 0.3 mg per g of cell dry weight. It is concluded that mycobactin biosynthesis is probably strongly repressed in the mycobactin-dependent strains rather than being a genetic deletion. The exochelins, when examined by high-pressure thin-layer chromatography were revealed as being multiples of similar compounds, with up to 20 individual iron-binding compounds being recognizable with some strains. It is argued that the exochelins represent the single most important means of iron acquisition in mycobacteria growing in vitro and in vivo, and their elaboration by the fastidious M. paratuberculosis and related species explains how these organisms are able to grow in vivo in the absence of an external supply of mycobactin.

Chromatography, Thin Layer↗
PubMed1990

T-cellular immune reactions (in macrophage inhibition factor assay) against Mycobacterium paratuberculosis, Mycobacterium kansasii, Mycobacterium tuberculosis, Mycobacterium avium in patients with chronic inflammatory bowel disease.

A mycobacterial aetiology has been suggested for Crohn's disease. A slow growing mycobacterium, biochemically and genetically identical to M paratuberculosis, the causative agent of enteritis in ruminants (Johne's disease), has been isolated from gut specimens of patients affected by Crohn's disease. If M paratuberculosis or other mycobacteria play a role in the pathogenesis of Crohn's disease, then patients may have been sensitised to these mycobacteria or show an anergy immune reaction. We therefore investigated the T-cell mediated immune response to sonicates of M paratuberculosis, M kansasii, M avium, and M tuberculosis in 35 patients with Crohn's disease, 28 with ulcerative colitis, and 25 controls using a macrophage inhibition factor assay on peripheral blood lymphocytes. Two types of reaction patterns were identified--that is, 'responders' (subjects with a macrophage inhibition factor assay in which a dose response relation was present and a percentage of inhibition exceeding 20%), and 'non-responders'. There was no significant difference in the prevalence of responders (59%-80%) and non-responders (20%-41%) to these mycobacteria between the group of Crohn's disease, ulcerative colitis, and control group. We found also that a large proportion of controls showed T-cell immunisation to the mycobacteria which supports the contention that the antigens are practically commensal. Our results do not support the proposed involvement of mycobacteria in the pathogenesis of Crohn's disease.

Adult↗
PubMed1986

Biochemical characteristics of various strains of Mycobacterium paratuberculosis.

Biochemical activities of 20 wild-type strains and of 2 laboratory strains of Mycobacterium paratuberculosis were evaluated. Biochemical activities evaluated were growth at 30 C, 37 C, and 42 C; production of urease, niacin, pyrazinamidase, arylsulfatase, and catalase; hydrolyzation of Tween 80; reduction of nitrate and tellurite; and growth in 5% NaCl. Antimicrobial susceptibility to thiophene-2-carboxylic acid hydrazide (10 micrograms/ml), neotetrazolium chloride (1:40,000), streptomycin (2 micrograms/ml), rifampin (0.25 micrograms/ml), and isoniazid (10 micrograms/ml) also was determined. Generally, M paratuberculosis was biochemically inactive, with only a few strains producing pyrazinamidase and maintaining catalase activity after heating. All strains grew optimally at 37 C, grew slightly at 30 C, and did not grow at 42 C. Wild-type strains did not grow in the presence of neotetrazolium chloride, streptomycin, and rifampin, and grew in the presence of thiophene-2-carboxylic acid hydrazide and isoniazid. Although biochemical evaluation can be used as an aid in the identification of M paratuberculosis, growth rate, and mycobactin dependency remain major criteria for positive identification.

Amidohydrolases↗