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PubMed1992

Effects of dialyzable lymph node extracts on lymphoblast proliferative capacity of blood mononuclear cells in cattle with chronic paratuberculosis.

Dialyzable lymph node extracts (DLE) containing transfer factor prepared from calves sensitized to Mycobacterium paratuberculosis and keyhole-limpet hemocyanin (KLH) were administered to 4 adult cows with chronic paratuberculosis. Cutaneous delayed hypersensitivity, lymphocyte blastogenesis, monocyte migration-inhibition, and lymphoblast proliferative capacity as a reflection of interleukin-2 (IL-2) activity were measured in response to M bovis purified protein derivative, johnin, and KLH before and after treatment with DLE. Change in cutaneous delayed hypersensitivity was not evident after DLE treatment. Alterations in histologic features of pre- and posttreatment sections of ileum and mesenteric lymph nodes were not detected. Lymph node extract treatment significantly (P less than 0.05) increased IL-2 activity and migration-inhibition in response to johnin and KLH in vitro. Treatment had no effect on lymphocyte blastogenesis. The data indicate that cattle with chronic paratuberculosis may benefit from DLE treatment, by virtue of increased IL-2 activity, and that effects of DLE are at least partially mediated by an increase in IL-2 activity.

Animals↗
PubMed1992

Measurement of lymphoblast proliferative capacity of stimulated blood mononuclear cells from cattle with chronic paratuberculosis.

Concanavalin A (conA) blast proliferation as a quantitative measure of lymphoblast proliferative capacity by blood mononuclear cell supernatants was measured in cattle naturally infected with Mycobacterium paratuberculosis and in healthy control cattle. Blast cell proliferation was significantly reduced in infected animals, compared with control cattle when blood mononuclear cells were stimulated with conA. Proliferation was significantly greater than media control when M bovis purified protein derivative and johnin were used to stimulate cells from the infected group. After sensitizing control and affected cattle with M paratuberculosis bacterin (live M bovis and keyhole limpet hemocyanin in Freund's incomplete adjuvant), infected animals had no difference in blast cell proliferative capacity with the mycobacterial antigens and conA stimulation, whereas healthy animals had significantly increased blast proliferation in response to all the sensitizing antigens. The blast cell proliferative capacity in infected animals with keyhole limpet hemocyanin stimulation was increased significantly after sensitization; however, it remained significantly less than that in the sensitized control group. These data indicate that cattle naturally infected with M paratuberculosis probably produce suboptimal interleukin-2 (IL-2) activity in response to a potent IL-2 inducer (conA) and fail to optimize IL-2 activity when sensitized with a potent immunogen (keyhole limpet hemocyanin).

Animals↗
PubMed1991

Enzyme-linked immunosorbent assay for detection of antibodies against Mycobacterium paratuberculosis in goats.

Using a heat and sonicated Mycobacterium paratuberculosis Cordoba antigen (COA1) and the commercial protoplasmic-antigen (PPA-3) as antigens, an ELISA for detecting goat antibodies was standardized. When 2 reference populations, 1 positive (17 goats) and the other negative (63 goats) to disease, were used, this test showed 87.5% sensitivity and 93.6% specificity for COA1, and 88.2 and 95.2%, respectively for PPA-3. Absorption with M phlei was performed; no significant differences were found for COA1, but a lower sensitivity was found with PPA-3. This test was not especially affected by cross-reactivity with other mycobacterial disease because when 9 goats with M bovis infection were included in the M paratuberculosis control group, the specificity was only slightly different for absorbed (94.4%) and nonabsorbed sera (91.7%) for COA1, and (93.1 and 94.4%, respectively) for PPA-3. This test was used to study the percentage of seropositive goats for M paratuberculosis in 3 herds with different prevalences. Among 251 goats in southern Spain (Huelva), 40% were found positive for COA1 and 41% for PPA-3. Among 242 goats studied in southern Spain (Córdoba), 10.0% were positive for COA1 and 13.0% for PPA-3. In the Canary Island population of 176 goats, 3% were positive for COA1 and 0.5% for PPA-3. According to the accuracies of both positive and negative predictions, our test could be applied to populations with high prevalence to prevent additions to the herd and to cull infected animals (with 40% prevalence, the positive and negative predictive values are 90%), and to prevent adding infected animals to populations with moderate or low prevalence.

Animals↗
PubMed1991

An evaluation of selected screening tests for bovine paratuberculosis.

The objective of this study was to evaluate the performance of the lipoarabinomannan antigen enzyme-linked immunosorbent assay (LAM-ELISA), carbohydrate antigen complement fixation (CH-CFT), and protein D antigen agar gel immunodiffusion (D-AGID) tests for bovine paratuberculosis, relative to histopathology, and to culture and isolation of Mycobacterium paratuberculosis from tissues and feces. Samples for test evaluation were collected from four sources including blood and tissues from 400 cull cows at three abattoirs in Ontario, blood and feces from a paratuberculosis survey of cattle from 120 dairy farms in Ontario, a serum bank containing samples from cattle from Ontario and Québec, and a bank of sera from cattle from Pennsylvania and the northeastern United States. The data were analyzed using receiver operator characteristic curves, estimates of relative sensitivity and specificity, and kappa statistics of agreement between tests. The LAM-ELISA performed significantly better than both the CH-CFT and the D-AGID tests. The LAM-ELISA was better at predicting fecal shedding status than tissue infection. However, the LAM-ELISA also had limitations. When interpreted as positive or negative (+/-), at a critical optical density of 0.675, its sensitivity and specificity relative to bacteriology were 49% and 87% respectively. Although the serological tests examined in this study provided some information, they did not predict well the infection status of individual animals.

Animals↗
PubMed1990

Gas chromatographic characterization of the relationship between some Mycobacterium avium and Mycobacterium paratuberculosis strains.

Mycobacterium avium strain P-55 and M. avium strain DENT differ from M. avium strain 16909-338 on the basis of their fatty acid spectra (C14:0, C18:0 and tuberculostearic [TBS] acids) studied by multivariate statistical analyses. Strains P-55 and DENT are closer to M. paratuberculosis strain 5889 than to M. avium strain 16909-338, a finding which is in harmony with earlier immunological observations. The recently isolated M. paratuberculosis strain 385 has proved different from M. paratuberculosis strain 5889.

Animals↗
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↗
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↗
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↗