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detection of antibodies to mycobacterium johnei by counterimmunoelectrophoresis.counterimmunoelectrophoresis (cie) was applied in the detection of antibodies to mycobacterium johnei in 110 sheep, 11 goat and 31 cattle sera and compared to immunodiffusion (id) test. one per cent noble agar, 7 ml per slide of 5 cm x 10 cm; barbitone-tris buffer, mu = 0.03, ph 8.6; a constant current of 5 ma per slide and m johnei protoplasmic antigen at 4 mg per ml were found to impart high sensitivity to cie and give rapid results. cie detected 97 sheep, 11 goat and 31 cattle positive sera, ...197896579
comparison of techniques for measuring the local and systemic responses to tuberculin in cattle.thermography of tuberculin skin reactions in cattle sensitized with mycobacterium avium, mycobacterium bovis, or mycobacterium paratuberculosis was rapid and sensitive, but was not as specifically related to the homologous sensitization as was increased skin thickness at 48 and 72 hours. systemic temperature responses were more completely monitored by radiotelemetry of intraperitoneally located temperature transmitters than by rectal thermometry, but the temperatures indicated by the 2 methods w ...1977319714
survival of mycobacterium paratuberculosis in slurry.cattle and swine slurry and a mixture of equal parts of both, was mixed with a culture of m. paratuberculosis, 0.1 mg per ml (1 mg = 33 x 10(6) viable units) and stored under anaerobic conditions at 5 degrees and 15 degrees c. at 5 degrees c the survival time for m. paratuberculosis was 252 days in all 3 kinds of slurry, and at 15 degrees c it was 98 days in cattle slurry, 182 days in swine slurry, and 168 days in mixed slurry.1977331238
skin testing, fecal culture, and lymphocyte immunostimulation in cattle inoculated with mycobacterium paratuberculosis.fourteen calves at 21 days of age were experimentally inoculated with 100 mg (wet weight) of mycobacterium paratuberculosis. three calves were inoculated orally, 4 intravenously, and 7 subcutaneously. lymphocyte immunostimulation, fecal culture, and intradermal tuberculin skin testing were done between 112 to 150 days following exposure. lymphocyte immunostimulation test results, conducted at 112 days after inoculation, showed all animals positive to mycobacterium avium purified protein derivati ...1977339793
immunological reactions caused by infection with mycobacterium paratuberculosis. a review.the natural resistance of cattle to infection with mycobacterium paratuberculsis in relation to their immunological reactions is recorded. in vivo and in vitro manifestations of cell-mediated immunity during infection are reviewed, together with the antibody responses. it is suggested that three distinct immunological states may arise following exposure to infection.1978351550
cultural and biochemical characteristics of mycobacterium paratuberculosis isolated from goats in norway. 1979375704
isolation of mycobacterium paratuberculosis from sheep and cattle in iceland. 1979384773
in vitro lymphocyte transformation as a herd survey method for bovine paratuberculosis.the lymphocyte-transformation (lt) test was evaluated for its potential application as a field test for bovine paratuberculosis. using a whole blood technique, samples from 3 consecutive collection periods were subjected to 3 mycobacterial antigens and to phytohemagglutinin. the results obtained from lt were compared with conventional serologic and cultural methods. a positive lt response to johnin purified-protein derivative (ppd) or avian ppd (or both) was noted in 40% to 60% of the animals te ...1978417649
culture of mycobacterium paratuberculosis from bovine foetuses. 1979464947
paratuberculosis (johne's disease) in bighorn sheep and a rocky mountain goat in colorado.between may, 1972 and february, 1978, six cases of paratuberculosis (johne's disease) caused by mycobacterium paratuberculosis were diagnosed in free-ranging rocky mountain bighorn sheep (ovis canadensis) and one rocky mountain goat (oreamnos americanus) on or near mt. evans in colorado. diagnosis of paratuberculosis was based on gross and histopathologic examination of the animals and by isolation of m. paratuberculosis from three sheep and the goat. the clinical signs and pathologic changes se ...1979480512
paratuberculosis in cattle and free-living exotic deer.paratuberculosis was studied among dairy cows and exotic deer that shared grazing areas at point reyes national seashore, california. of the 10 dairy herds tested, 5 (50%) were infected with mycobacterium paratuberculosis (based on results of fecal culture). mycobacterium paratuberculosis was cultured from 9 (8.7%) of the 103 bovine fecal samples and from 4 (3.9%) of the 103 bovine rectal mucosa scapings tested. of 89 fecal samples from 52 axis deer (axis axis) and 37 fallow deer (dama dama), 5 ...1979500426
observations on diagnostic tests for paratuberculosis in a deer herd.paratuberculosis was diagnosed in a herd of 62 fallow deer (dama dama) and 38 sika deer (cervus nipon) maintained in an animal park in ohio. a comparison was made of delayed-type hypersensitivity responses, results of in vitro lymphocyte immunostimulation tests, and isolation of mycobacterium paratuberculosis on fecal culture in 19 deer.1979521371
susceptibility of the lemming (dicrostonyx rubricatus) to mycobacterium paratuberculosis.the susceptibility of the lemming (dicrostonyx rubricatus) to mycobacterium paratuberculosis was studied. twelve lemmings were exposed to cultural material of m paratuberculosis and were necropsied at 40- to 260-day intervals after exposure. selected tissues were examined bacteriologically and histologically. mycobacterium paratuberculosis was recovered from all exposed lemmings except the one necropsied 40 days after exposure. pathologic changes typical of paratuberculosis were seen in a few le ...1979525886
the prevalence of antibodies to mycobacterium johnei in colostrum deprived lambs.the prevalence of antibodies to mycobacterium johnei in pre-colostral lambs in a flock of sheep infected with johne's disease was investigated. blood from 134 ewes and their 143 lambs were collected immediately after lambing. double immunodiffusion and counterimmunoelectrophoresis tests were used to detect antibodies to m johnei. four (3 per cent) of 135 lambs born to 127 ewes with serological evidence of exposure to m johnei had detectable antibodies in their sera. all lambs fed colostrum from ...1979552705
radiommunoassay for antibodies against mycobacterium paratuberculosis using 125i-labeled ppd. 1978566028
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 reac ...1977596700
johne's disease and the practitioner.mycobacterium paratuberculosis infections and the disease states resulting in cattle are defined as spectral in nature. there is a spectrum of immunological response which complicates tests based on this. it is recommended that initial diagnosis of suspicious clinical cases be confirmed by direct fecal smears and a serological test such as immunodiffusion. herd evaluation requires the application of fecal cultures plus immunodiffusion. control requires rigid management hygiene measures, surveill ...1978630894
pathological evaluation of paratuberculosis in naturally infected cattle.thirty-two of 51 cattle infected with mycobacterium paratuberculosis had chronic enteritis, chronic lymphangitis or mesenteric lymphadenopathy, or all three, at slaughter. granulomatous inflammatory lesions were mild to advanced and predominantly involved the distal small intestine. rectal involvement was seen only in five cattle. fourteen had microgranulomas in the liver. there were three cytological forms of macrophages: histiocytic, polygonal and epithelioid. the latter two types had engulfed ...1978664186
enzyme-linked immunosorbent assay (elisa) for detection of antibodies to mycobacterium paratuberculosis in cattle. 1978696542
age and milk production data of cattle culled from a dairy herd with paratuberculosis.statistical assessment of age and milk production data revealed a significantly shorter life expectancy and reduced milk production of mycobacterium paratuberculosis-infected dairy cows, when compared with non-infected herdmates. high producing cows were frequently culled after their 1st or 2nd gestation, contributing to an undetermined economic loss with regard to their potential breeding value. cows with subclinical infection frequently had problems of infertility and of mastitis.1978711590
experimentally induced infectious bovine keratoconjunctivitis: potentiation of a moraxella bovis pilus vaccine's immunogenicity by vaccination with mycobacterium paratuberculosis bacterin. 1978717879
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 ...1978736348
serological similarity between caprine and bovine strains of mycobacterium paratuberculosis. 1978743046
mammary gland exposure of cows to mycobacterium paratuberculosis.mycobacterium paratuberculosis injected into the mammary gland was transported to the supramammary lymph nodes in 5 of 6 cows and to the intestine of 1 cow. the bacillus caused hypersensitivity to johnin and stimulated the production of complement-fixing antibodies. apparently, the bacillus is not a cause mastitis.1978749582
the specificity and sensitivity of the fluorescent antibody test for mycobacterium johnei infection in abattoir and culled cattle.in a series of 100 cattle in which there was no bacteriological or histopathological evidence of mycobacterium johnei infection there were four positive reactions to the fluorescent antibody (fa) test with m johnei antigen and 20 to the complement fixation (cf) test. in a second series of 118 culled adult cattle m johnei infection was found in 26. the fa test was positive in 16 and the cf in 12 of those infected cattle. in 92 cattle with no evidence of m johnei infection the fa test was positive ...1976769122
the specificity and sensitivity of the fluorescent antibody test in cattle experimentally infected with mycobacterium avium and mycobacterium johnei.in five calves experimentally infected with mycobacterium avium and in 10 with m johnei it was shown that the serological response by the fluorescent antibody (fa) test was specific. the serological response in the m avium infected calves was transitory and lasted up to five months. the first evidence of a serological response with the fa test in the m johnei dosed calves occurred at four months, and all calves had reacted by nine months after dosing. at the 23 tests carried out between four and ...1976769123
diagnosis of johne's disease in cattle: a comparison of five serological tests under field conditions.sera from 775 adult milking cattle on 13 farms in friesland and from 359 of the same cattle taken during the following year, were examined for evidence of infection with m johnei by the complement fixation, fluorescent antibody, haemagglutination, haemagglutination-lysis and immunodiffusion tests. eighteen animals, not clinically affected but which had serologically positive tests were slaughtered and m johnei was isolated in cultures from 16. the fluorescent antibody test was as sensitive as th ...1976820056
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 ...1977911084
effect of freezing on the viability of mycobacterium paratuberculosis in bovine feces.three bovine fecal specimens were cultured for mycobacterium paratuberculosis before freezing and after frozen storage at -70 degrees c for 3 and 15 weeks. the losses in viability from 0 to 3 weeks of storage were significant (p = 0.01) for all three samples. the losses in viability between 3 and 15 weeks of storage were not significant (p greater than 0.05) for two specimens in which the m. paratuberculosis occurred naturally, but the loss was significant (p = 0.01) in a simulated specimen (m. ...1977914994
subcutaneous exposure of calves to myobacterium paratuberculosis compared with intravenous and oral exposures.three groups of calves, 21 days of age, were exposed to mycobacterium paratuberculosis by oral, intravenous, and subcutaneous routes. fecal cultural examination and intradermal tests were made, and at 150 days after exposure, the calves were euthanatized and tissues were collected for histopathologic and bacteriologic cultural examination. calves in the 3 groups were infected. those exposed intravenously had significantly higher colony counts than did either of the other groups. there was no sig ...1977931152
susceptibility of the mongolian gerbil (meriones unguiculatus) to mycobacterium paratuberculosis.mongolian gerbils (meriones unguiculatus) were exposed to mycobacterium paratuberculosis by intraperitoneal and oral routes. fourteen weeks after exposure, the gerbils were necropsied and tissues were cultured and examined microscopically. results of the study showed that gerbils can be infected with m paratuberculosis but are not suitable for studies of paratuberculosis.1976962215
[several strains of mycobacterium paratuberculosis: biochemical characteristics and allergic activity].judged from its biochemical properties and antigenic structure, mycobacterium paratuberculosis is related to m. avium. when m. paratuberculosis was compared to b.c.g., several essential differences, were found that allowed their clear separation, within runyon's group iii, m. paratuberculosis is differentiated by its biochemical properties as shown in the following table : (formula: see text). the allergic response of a guinea pig infected with m. paratuberculosis was stronger when an avium sens ...19761023808
isolation and identification of mycobacteria from porcine tissues: a three-year summary.mycobacteria were isolated from 1,591 (78%) of 2,036 porcine tissues submitted to veterinary services laboratories over a 3-year period (july 1, 1971, to june 30, 1974). the isolates were identified by biochemical and serologic tests. of the 1,547 mycobacterium avium isolates, 452 were serotype 1, 728 were serotype 2, 60 were serotyped 4, 110 were serotype 8, and 51 were serotyped 10; 36 isolates represented 11 other serotypes; 65 isolates shared antigens with more than one serotype; and 45 isol ...19751099946
age of cattle as related to resistance to infection with mycobacterium paratuberculosis.two 1-month-old calves, four 9-month old calves, and 4 adult (5 to 11 years old) cattle were exposed to mycobacterium paratuberculosis. at 150 days after exposure, all cattle were killed and necropsied, and at that time, tissues of the 1-month-old calves had more bacilli and lesions than did tissues of the 9-month-old calves or the adult cattle. all cattle responded to immunologic tests during the experiment.19751115424
analysis of the effect of inapparent bovine paratuberculosis.inapparent paratuberculosis was studied in a guernsey herd with a history of paratuberculosis (johne's disease); the herd was maintained at between 900 and 950 cattle. fecal specimens and intestinal tissue specimens from any of these that were slaughtered were examined culturally for the presence of mycobacterium paratuberculosis. the reasons given for culling and slaughtering of cows from this herd were compared with infection status determined by culturing. less than one-third of the cultural ...19751147338
seroagglutination test for identification of mycobacterium paratuberculosis.twenty-nine of 30 strains of mycobacterium paratuberculosis isolated from clinical specimens were serologically identified. no cross-reactions were observed with mycobacterium avium serotypes 1, 2, 4, and 8.19751176634
effect of aflatoxin on susceptibility of hamsters to mycobacterium paratuberculosis.after oral administration of mycobacterium paratuberculosis to hamsters, the organism passed the epithelial barrier of the intestine, and infection was established in the small intestine and mesenteric lymph nodes. the addition of aflatoxin to the ration of hamsters did not increase their susceptibility to m paratuberculosis but, rather, seemed to decrease susceptibility to the bacillus. hamsters not treated with aflatoxin and infected with m paratuberculosis had higher bacterial counts in intes ...19751190596
intravenous johnin and tuberculin tests in cattle vaccinated with mycobacterium paratuberculosis cells and subsequently inoculated with mycobacterium bovis.calves at 30 days of age were vaccinated with a killed whole-cell mycobacterium paratuberculosis vaccine. four months later, these calves were inoculated with mycobacterium bovis. the intravenous tuberculin and johnin tests were applied both before and after inoculation. the results of the hematologic investigation had extremes at both high and low values and were too unsuitable for statistical analysis. the intravenous tuberculin test is considered unsuitable for diagnosis of bovine tuberculosi ...19751200443
antibodies to mycobacterium paratuberculosis-specific protein antigens in crohn's disease.the possible role of infection with mycobacterium paratuberculosis (map) for the etiopathogenesis of crohn's disease (cd) has been a matter of long-term controversy. in addition to similarities with the pathology of ruminant paratuberculosis, dna fingerprinting confirmed the organism isolated from gut tissue, but the specificity of the immune repertoire has not as yet been evaluated. we report here on a serological study of 29 patients with cd, 20 patients with ulcerative colitis and 18 healthy ...19921281056
paratuberculosis vaccine in a large dairy herd.on a 500-cow dairy farm a total of 866 young calves less than one month old were vaccinated with a heat-killed oil-adjuvated bacterin against mycobacterium paratuberculosis over a period of five years. the vaccinated calves were tested by faecal microscopy, bacteriology and serology on the day of vaccination, at the age of 3, 6, 9 and 12 months, at breeding age, and on the day of calving. a total of 721 bull calves and 379 female calves served as unvaccinated controls in two groups. the results ...19921298162
the use of antimycobacterial agents in crohn's disease.seventy-five years ago, a scottish surgeon, dalziel, clearly described crohn's disease (cd) and suggested that it might be caused by a mycobacteria. however, mycobacteria were not isolated from cd tissue until 1978 and 1984. since then several investigators, using sophisticated polymerase chain reaction (pcr) techniques, found mycobacterium paratuberculosis sequences in cultures of crohn's disease tissue in approximately two thirds of patients. because of a possible mycobacterial etiology, rutge ...19921323594
characterization of ovine strains of mycobacterium paratuberculosis by restriction endonuclease analysis and dna hybridization.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.19921325047
p43, the protein product of the atypical insertion sequence is900, is expressed in mycobacterium paratuberculosis.the novel mycobacterial insertion sequence is900 was analysed by coupled transcription-translation, of both strands independently, in a cell-free e. coli extract using an exogenous promoter. this revealed only one protein product, p43, as predicted from the nucleotide sequence. the protein was readily translated in recombinant e. coli, using the tac promoter, though it did not appear as a major product by sds-page analysis. a synthetic peptide was used to generate and affinity-purify a specific ...19921326596
biologically distinct subtypes of mycobacterium avium differ in possession of insertion sequence is901.mycobacterium avium causes disease, principally tuberculosis in immunocompromised individuals. it is the most frequent cause of disseminated infections in aids patients in the west. the pathogen is also associated with disease in animals, chiefly birds and livestock, and may be isolated from environmental samples such as soil and water. analysis of strains of m. avium isolated from clinical, veterinary, and environmental sources for the presence of the mycobacterial insertion sequences is900 and ...19921328288
immunosuppression in goats by dexamethasone and cyclophosphamide.the influence of dexamethasone and cyclophosphamide on the goat immune system was investigated. seven goats, with a previous contact with caprine herpesvirus type 1 (chv-1), were used. all had been vaccinated with live mycobacterium paratuberculosis vaccine. six goats were injected intravenously (i.v.) with dexamethasone daily for 5 days (2.5-4 mg/kg bw per day). three also received 25 mg/kg bw of cyclophosphamide on day 0. the seventh goat was not treated. dexamethasone alone caused depression, ...19921330422
expression of escherichia coli beta-galactosidase in mycobacterium bovis bcg using an expression system isolated from mycobacterium paratuberculosis which induced humoral and cellular immune responses.a promoter sequence, pan, was isolated from mycobacterium paratuberculosis and characterized. this promoter lies adjacent to, and outside, the 3' end of an is900 insertion element. is900 contains an open reading frame, orf2, on the complementary strand which codes for the putative transposase of this insertion sequence. a dna fragment containing pan and part of orf2 was fused to the lacz gene and inserted into the replicative shuttle vector prr3. mycobacterium smegmatis and mycobacterium bovis b ...19921336563
is902, an insertion element of the chronic-enteritis-causing mycobacterium avium subsp. silvaticum.an insertion sequence element of mycobacterium avium subsp. silvaticum was isolated and its complete nucleotide sequence determined. is902 is 1470 bp in size and is repeated 10-12 times per genome. an open reading frame of 1200 bp was identified, encoding a protein product of mr 43932. this protein is highly similar to the predicted proteins of is900 of mycobacterium paratuberculosis, is116 of streptomyces clavuligerus and is110 of streptomyces coelicolor. is902 lacks terminal inverted repeats a ...19921348267
use of restriction fragment length polymorphisms resolved by pulsed-field gel electrophoresis for subspecies identification of mycobacteria in the mycobacterium avium complex and for isolation of dna probes.mycobacterial strains from the mycobacterium avium complex were compared with each other and with mycobacterium phlei isolates by restriction endonuclease digestion of chromosomal dna with sspi and analysis by pulsed-field gel electrophoresis. characteristic profiles were observed for known typed strains, and five groups were identified. primary bovine isolates identified as mycobacterium paratuberculosis by classical methods were shown to fall into both the m. paratuberculosis- and m. avium-lik ...19921352787
mycobacteria in crohn's disease: dna probes identify the wood pigeon strain of mycobacterium avium and mycobacterium paratuberculosis from human tissue.mycobacterium paratuberculosis is known to cause johne's disease, a granulomatous ileitis in ruminants, and may be involved in some cases of crohn's disease. like m. paratuberculosis, the wood pigeon strain of mycobacterium avium may also show mycobactin dependence on primary isolation that is attenuated on further subculturing. a wood pigeon strain, m. avium restriction fragment length polymorphism (rflp) type a/i, is also capable of causing granulomatous ileitis in experimental animal models b ...19921360477
the cellular immunology of bovine paratuberculosis: the predominant response is mediated by cytotoxic gamma/delta t lymphocytes which prevent cd4+ activity.peripheral blood t-cell subsets were obtained from an experimentally sensitized bovine and from nine bovines naturally infected with mycobacterium paratuberculosis and tested for their ability to respond to antigen. it was determined that following antigen challenge, proliferative responses followed a biphasic pattern: an initial cd4+ proliferative response, a period of anergy, and a final response governed by gamma/delta t lymphocytes. the anergic phase was characterized by a dramatic drop in p ...19921363823
amplification of 16s rrna sequences to detect mycobacterium paratuberculosis.a probe based on 16s ribosomal rna (rrna) sequences was developed to detect mycobacterium paratuberculosis, the causative agent of johne's disease in cattle. three universal primers were used to sequence the amplified fragments of the 16s rrna gene of various species of mycobacteria. when the nucleotide sequences were analysed, a deletion was detected in the sequence of the fast-growing species. an oligonucleotide probe (p) directed to this region was synthesised and hybridised directly with tot ...19921373192
composition and immunological properties of the protein fraction of a36, a major antigen complex of mycobacterium paratuberculosis.tma (thermostable macromolecular antigens) are major mycobacterial complexes present in all mycobacteria. we have purified a36, the tma complex of m. paratuberculosis, the etiological agent of paratuberculosis (johne's disease), and shown by the immune electron microscopy approach its presentation at the cell surface. the immunodominance of the a36 complex in johne's disease was suggested by comparative elisa analysis of infected bovine sera, using either a36 or m. paratuberculosis total soluble ...19921380177
experimental infection of severe combined immunodeficient beige mice with mycobacterium paratuberculosis of bovine origin.severe combined immunodeficient beige mice were inoculated orally and intraperitoneally with a bovine strain of mycobacterium paratuberculosis to explore their potential as laboratory animal models in the study of paratuberculosis (johne's disease). control animals were similarly inoculated with heat-killed m. paratuberculosis. in the mice inoculated intraperitoneally, focal lesions and acid-fast bacilli were first detected in the livers (4 weeks postinfection) and later in the spleens and intes ...19921398920
detection of mycobacterium paratuberculosis antigen with colloidal immunogold in naturally infected sheep.the anti-mycobacterium paratuberculosis polyclonal serum is proved useful for labelling mycobacterium paratuberculosis in glutaraldehyde-osmium-fixed and epon-embedded intestinal samples from sheep with clinical symptoms of paratuberculosis. m. paratuberculosis marked with antibody-coated colloidal gold stain was seen in macrophages, epithelioid cells, giant cells and neutrophils throughout intestinal mucosa. in large macrophages with a low lysosomal content, a great number of intact mycobacteri ...19921414096
polymerase chain reaction detection of mycobacterium paratuberculosis and mycobacterium avium subsp silvaticum in long term cultures from crohn's disease and control tissues.thirty one cultures were established in mg3 medium from the intestinal tissues of 29 patients, including 18 with crohn's disease, five with ulcerative colitis, and six non-inflammatory bowel disease controls. all cultures grew either acid fast bacilli or uncharacterized spheroplasts. pellets from these cultures were coded and assayed blind for m paratuberculosis and m avium subsp silvaticum using is900- and is902-pcr (polymerase chain reaction) assays, respectively. is900 and is902 are multicopy ...19921427373
molecular cloning and characterization of mycobacterium paratuberculosis promoters in escherichia coli.dna fragments from mycobacterium paratuberculosis were cloned in the promoter probe plasmid pko1. of 957 recombinant dna clones, 24 induced synthesis of galactokinase (the reporter gene) when these plasmids were transformed into an escherichia coli strain deficient for the enzyme. a dna insert from one putative promoter-containing plasmid, designated pag5, was sequenced and shown to contain, a characteristic rna polymerase binding site, a probable ribosomal binding site and a putative open readi ...19921455629
induction of cellular immune reactions by a36, an antigen complex of mycobacterium paratuberculosis: comparison of a36 and johnin components.paratuberculosis (johne's disease) is a chronic enteritis syndrome of ruminants, which is due to infection by mycobacterium paratuberculosis. cutaneous testing with proteins extracted from a mycobacterial culture fluid (johnin-ppd) is currently used to evaluate the cellular immune status. we have compared the components of johnin-ppd with those of the a36 complex, a thermostable macromolecular antigen (tma) present in the cytoplasm and associated with the cell wall of m. paratuberculosis. the pr ...19921462120
activity of clarithromycin compared with those of other drugs against mycobacterium paratuberculosis and further enhancement of its extracellular and intracellular activities by ethambutol.radiometric mics of clarithromycin, a new macrolide drug, were determined against five mycobactin-dependent strains of mycobacterium paratuberculosis (including two crohn's disease clinical isolates) and compared with those of other drugs which included rifampin, ethambutol, amikacin, ofloxacin, ciprofloxacin, and sparfloxacin. among the drugs screened, clarithromycin was the drug for which mics were lowest against the five strains tested. as mics were significantly below the reported cmax level ...19921482157
protein antigens secreted by mycobacterium paratuberculosis.proteins secreted by mycobacterium paratuberculosis (m.ptb) during short-term cultivations were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page) and western(immuno) blotting. cultivation in a defined medium containing 35s methionine allowed autoradiographic detection of proteins which had been secreted or passively released by actively metabolizing m.ptb organisms. after the first 3 days of cultivation, 4 proteins with molecular weights of approximately 38, 50, 65 ...19921492518
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 ...19921497195
isolation of mycobacterium paratuberculosis after oral inoculation in uninfected cattle.feces from cows naturally infected with mycobacterium paratuberculosis was given to 6 uninfected heifers by orogastric intubation, to determine whether ingested organisms could be passively excreted and detected by bacteriologic culture of feces (ie, false-positive result). heifers were paired, and each pair received a different dose of feces on days 1 and 2. fecal samples were collected from the heifers 3 times daily. mycobacterium paratuberculosis was detected in fecal samples of all heifers w ...19921510305
comparison of subjective and objective test evaluations for use of mycobacterium phlei-adsorbed serum in a dot-enzyme-linked immunosorbent assay for diagnosis of paratuberculosis in cattle.use of a dot-elisa with serum adsorbed with mycobacterium phlei or with nonadsorbed serum was compared. in addition, results attained using visual observation were compared with those obtained using a densitometer. infection status of cattle was determined by results of culture of feces from a number of cattle with various degrees of exposure (low prevalence and test-negative) and disease manifestation (clinical suspect vs subclinical infection). two paratuberculosis-negative herds, fecal cultur ...19921510315
selection for immune response in goats: the effect of immunization procedure on antibody response to diphtheria toxoid and human serum albumin.the serum antibody titers to diphtheria toxoid and human serum albumin were determined in 103 goat kids from lines selected for 12 yr for high or low antibody response to diphtheria toxoid. in the 12th yr, six groups of kids were immunized with different preparations of the antigens. in all groups but one, the antigens were emulsified in freund's incomplete adjuvant with added sonicated mycobacterium paratuberculosis. the groups received the following treatments: group 1 was immunized with both ...19921526911
antigen induced suppression in peripheral blood and lamina propria mononuclear cells in inflammatory bowel disease.using an autologous system, suppressor cell function to a range of mycobacterial antigens and kunin antigen of peripheral blood mononuclear cells and lamina propria lymphocytes has been investigated in normal subjects and patients with inflammatory bowel disease. in the peripheral blood there was reduced antigen induced suppression in patients with crohn's disease in remission to mycobacterium paratuberculosis, purified protein derivative (ppd), m fortuitum, and kunin antigens (p less than 0.05) ...19921533199
diagnostic accuracy of a mycobacterium phlei-absorbed serum enzyme-linked immunosorbent assay for diagnosis of bovine paratuberculosis in dairy cows.the purpose of this study was to describe the responses of sera from five groups of cattle to an enzyme-linked immunosorbent assay (elisa) for paratuberculosis by using serum absorbed with mycobacterium phlei at a single working dilution. the infection status of the cattle was determined by fecal culture. cattle with different levels of exposure (high versus low prevalence and test negative) and disease manifestation (clinically suspect infection versus subclinical infection) were examined, as f ...19921551978
comparison of a commercial dna probe test and three cultivation procedures for detection of mycobacterium paratuberculosis in bovine feces.diagnosis of paratuberculosis using the idexx dna probe test and 3 methods for cultivation of mycobacterium paratuberculosis from fecal specimens were compared. twenty-one of 170 fecal specimens were dna probe test positive, whereas 35 specimens were positive by 1 or more of the cultivation methods evaluated. four specimens were dna probe test positive but were negative by fecal culture. the probe test detected m. paratuberculosis dna in 62.9% of the specimens positive by a sedimentation culture ...19921554765
composition and immunogenicity of the polysaccharide components of the thermostable macromolecular antigen group of mycobacterial antigens.the thermostable macromolecular antigen (tma) group includes major components of the mycobacterial cell envelope and cytoplasm, which elicit humoral and cellular immune reactions, and seems to play important roles in infectious diseases. the best known member of this group, antigen a60 of mycobacterium bovis bcg, was previously shown to contain three moieties of polysaccharides, free lipids, and polypeptides. in this work, the tma polysaccharides of three pathogenic mycobacteria (m. avium, m. bo ...19921579083
triple-helical capture assay for quantification of polymerase chain reaction products.this method for rapid, automated analysis of polymerase chain reaction (pcr) products makes use of pcr primers containing 5'-polypyrimidine sequences. polypyrimidine-"headed" primers confer to the pcr product the ability to form triple helical complexes with a third polypyrimidine oligonucleotide. third-strand oligonucleotides are modified to serve as either capture reagents or detection reagents for pcr products. automated quantitative measurement of the pcr product is achieved by using latex b ...19921582021
mycobacterium paratuberculosis. factors that influence mycobactin dependence.mycobacterium paratuberculosis does not produce any detectable mycobactin, an iron-binding compound that is synthesized by most mycobacterium spp. and necessary for the growth of all mycobacteria. this study examined the influence of various culture conditions on mycobactin dependence in m. paratuberculosis. using a radiometric growth assay, we found the minimal concentration of mycobactin j necessary for growth of m. paratuberculosis to be 0.006 microm, whereas 1.2 microm (1 microgram/ml) was r ...19921582168
evaluation of four serological tests for bovine paratuberculosis.the standard complement fixation (cf) test, a commercial agarose gel immunodiffusion (agid) test (immucell corporation, portland, maine), and two commercial enzyme-linked immunosorbent assays (elisas; allied laboratories, glenwood springs, colo. [allied elisa], and the csl, limited, [parkville, victoria, australia] enzyme immunoassay [csl elisa]) were evaluated by using sera from the repository for paratuberculosis specimens. the case definition of subclinical bovine paratuberculosis was isolati ...19921583110
evaluation of the abilities of three diagnostic tests based on the polymerase chain reaction to detect mycobacterium paratuberculosis in cattle: application in a control program.three assays for the specific detection of mycobacterium paratuberculosis by dot spot hybridization of polymerase chain reaction products were applied to fecal samples of dairy cattle. the first two tests used polymerase chain reaction primers and a dna probe derived from m. paratuberculosis-specific sequences of the 16s rrna gene and insertion element is900, respectively. these two tests were carried out on spiked fecal samples to determine the detection limits. the 16s rrna test was able to de ...19921583122
mycobacterium paratuberculosis isolated from fetuses of infected cows not manifesting signs of the disease.fetuses were obtained from 58 cows that were fecal culture-positive for mycobacterium paratuberculosis, but were not manifesting signs of paratuberculosis. fetal tissues from 5 of 58 cows were culture-positive for m paratuberculosis. all 5 culture-positive fetuses were from cows that were classified as heavy fecal shedders (5/28; 17.8%). difference in prevalence of fetal infection between light (less than 70 colonies/tube) and heavy fecal shedders was significant (fisher's exact test, p less tha ...19921586015
prophylactic effect of monensin sodium against experimentally induced paratuberculosis in mice.monensin sodium (0, 15, or 30 mg/kg of complete feed) was fed ad libitum for 1 week to female mice (strain c57bl6/j) that were genetically susceptible to infection with mycobacterium paratuberculosis. ten mice in each of the 3 groups were inoculated intraperitoneally with m paratuberculosis (10(9) organisms). sterile saline solution was injected intraperitoneally into 10 other mice in each group. rations were continued for 50 days, then mice were euthanatized, and body weight, splenic weight, an ...19921586026
effects of mycobactin j and lactoferrin supplementation of drinking water on the in vivo multiplication of mycobacterium paratuberculosis in gnotobiotic mice.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.19921586898
evaluation of conventional and radiometric fecal culture and a commercial dna probe for diagnosis of mycobacterium paratuberculosis infections in cattle.radiometric (rcm) and conventional fecal culture (hey) and a commercial polymerase chain reaction/dna probe were evaluated as diagnostic tests for subclinical paratuberculosis in dairy cattle using fecal specimens from a repository of paratuberculosis specimens. the case definition of subclinical bovine paratuberculosis was isolation of mycobacterium paratuberculosis, by conventional or radiometric culture, from fecal samples or internal organs of dairy cattle without diarrhea or chronic weight ...19921591658
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 ...19921595967
paratuberculosis in saiga antelope (saiga tatarica) and experimental transmission to domestic sheep.mycobacterium paratuberculosis was isolated in low numbers from the small intestine and associated mesenteric lymph nodes of a saiga antelope (saiga tatarica) using routine culture techniques in spite of histologic evidence of high numbers of acid-fast bacteria in these tissues. two newborn domestic sheep were fed the ground intestinal tissue containing acid-fast bacteria and the progression of the experimental disease was followed by fecal culture, immunodiffusion (agid) and lymphocyte stimulat ...19921602565
[experimental paratuberculosis in sheep after intravenous or oral inoculation: pathogenicity and biologic diagnosis].an experimental paratuberculosis study was performed in sheep. one group of 6 lambs was inoculated intravenously with the equivalent of 50 mg (wet weight) of live bacilli, another group of 6 lambs was inoculated orally by placing 500 mg (wet weight) of live organisms in milk feed and a group of 3 lambs was used as controls. the degree of cellular immunity was followed by examining delayed hypersensitivity using 3 allergens (bovine tuberculin ppd, avian tuberculin ppd and johnine ppd) and that of ...19921610074
a repository of specimens for comparison of diagnostic testing procedures for bovine paratuberculosis. 19921616985
[bacterial studies on the occurrence of mycobacterium paratuberculosis in fecal samples of zoo ruminants].mycobacterium (m.) paratuberculosis was isolated from fecal samples of 3 (21.4%) from 14 mouflons, of 10 (20.4%) from 49 dwarf goats, of 5 (14.3%) from 35 cameroon sheep and of 1 (9.1%) from 11 alpine ibex. m. paratuberculosis could not found by cultural method in fecal samples of 22 pinzgauer goats, of 15 bantengs, of 9 wild goats, of 9 skuddens, of 6 four-horned sheep, of 3 red-head sheep, and of 1 chamois. from all 19 animals with cultural positive fecal samples complement binding antibodies ...19921622380
[current insights on the pathogenesis of chronic inflammatory bowel diseases].the pathogenesis and etiology of inflammatory bowel disease (ibd) is poorly understood. as a matter of fact, it is not even certain whether either one is a single entity with different forms of clinical manifestations, or whether each one represents a single clearly separable entity. common features of both diseases are chronic persistence, recurrent exacerbation and remission, the production of autoantibodies, as well as the expression of aberrant hla-class ii molecules on the surface of epithe ...19921626162
a comparison of the interferon gamma assay with the absorbed elisa for the diagnosis of johne's disease in cattle.two interferon gamma (ifn-gamma) assays, the ifn-gamma enzyme immunoassay (eia) and the ifn-gamma bioassay and an absorbed elisa were used to screen 6 cattle herds for johne's disease. each herd had a history of johne's disease but the majority of infected animals did not show clinical signs. the disease status of the cattle, which were removed from the herds, was confirmed by bacteriological culture of faeces or histopathological examination and culture of tissues collected at necropsy. the sen ...19921632724
mycobacterium paratuberculosis binds fibronectin.fibronectin, an adhesive glycoprotein which is present in plasma and on many host cell surfaces of many host organisms, binds to certain bacterial pathogens. this study demonstrates the ability of mycobacterium paratuberculosis (m.ptb) to interact with 125i-labelled fibronectin purified from bovine and ovine plasma. two m.ptb strains were tested: a clinical isolate and a commercially available vaccine strain. both strains showed significant fibronectin-binding activities of 22 and 41%, respectiv ...19921641514
mycobacterium paratuberculosis dna in crohn's disease tissue.crohn's disease has long been suspected of having a mycobacterial cause. mycobacterium paratuberculosis is a known cause of chronic enteritis in animals, including primates, but may be very difficult to detect by culture. is900 is a multicopy genomic dna insertion element highly specific for m paratuberculosis. a polymerase chain reaction (pcr) based on the 5' region of is900 and capable of the specific detection of a single m paratuberculosis genome was developed. this was applied to dna extrac ...19921644328
antimicrobial activity of rifabutin in combination with two and three other antimicrobial agents against strains of mycobacterium paratuberculosis.the inhibitory and bactericidal, synergistic, and antagonistic activities of rifabutin combined with ciprofloxacin, ethambutol, clofazimine, cefazolin, and amikacin in dual and triple combinations against various human and animal isolates of mycobacterium paratuberculosis were determined. synergism was observed when rifabutin was combined with either cefazolin or clofazimine in double combinations. the greatest amount of synergy occurred with the rifabutin-cefazolin combination in which bacteric ...19911647388
is901, a new member of a widespread class of atypical insertion sequences, is associated with pathogenicity in mycobacterium avium.an insertion sequence (is901), found in pathogenic strains of mycobacterium avium, but absent in m. avium complex isolates from patients with acquired immune deficiency syndrome (aids), has been isolated and sequenced. this insertion element has a nucleotide sequence of 1472 bp, with one open reading frame (orf1), which codes for a protein of 401 amino acids. the amino acid sequence, terminal ends and target site of is901 are similar to those of is900, present in mycobacterium paratuberculosis. ...19911685008
polyclonal antibodies raised against bacillus calmette-guerin, mycobacterium duvalii, and mycobacterium paratuberculosis used to detect mycobacteria in tissue with the use of immunohistochemical techniques.commercially available polyclonal antibodies raised against strains of mycobacteria were used to detect organisms in tissue sections from 34 cases of tuberculosis, leprosy, and atypical mycobacteria. thirty-two cases of fungal infections, granulomatous inflammation, and sarcoidosis were used as negative controls. sections stained with the use of antibodies raised against bacillus calmette-guerin (bcg), mycobacterium duvalii (md), and mycobacterium paratuberculosis (mp) were compared with kinyoun ...19901697733
discovery of an insertion sequence, is116, from streptomyces clavuligerus and its relatedness to other transposable elements from actinomycetes.we have identified an insertion sequence, is116, present in streptomyces clavuligerus at one copy per genome. the element was discovered as a 1.4 kb insertion into the multicopy plasmid pij702 after propagation in s. clavuligerus. the nucleotide sequence of is116 and the flanking sequences from pij702 have been determined. the junctions with pij702 show no target site duplication and there are no inverted repeats at the ends of the element. one putative coding open reading frame of 1197 bp was i ...19901700062
gen-probe rapid diagnostic system for distinguishing between mycobacterium avium and mycobacterium paratuberculosis. 19911719027
mycobacterium paratuberculosis cultured from milk and supramammary lymph nodes of infected asymptomatic cows.milk and supramammary lymph node samples were obtained from asymptomatic cows infected with mycobacterium paratuberculosis at the time of slaughter. of 81 supramammary lymph node samples, 22 (27%) were culture positive for m. paratuberculosis. of 77 milk samples, 9 (11.6%) were culture positive. the prevalence of supramammary lymph node or milk infection was highest with heavy fecal shedding of m. paratuberculosis and lowest with light shedding. the serologic status of the cow was not useful for ...19921734049
mucosal cell-mediated immunity to mycobacterial, enterobacterial and other microbial antigens in inflammatory bowel disease.culture studies have suggested that mycobacterium paratuberculosis may play a role in the aetiology of crohn's disease. however, evidence of sensitization to mycobacterial antigens amongst patients with crohn's disease has not yet been adequately demonstrated. previous studies of cell-mediated immunity (cmi) in crohn's disease were restricted to responses of peripheral blood mononuclear cells (pbmc) to mycobacterial antigens. in this study we have investigated the proliferative responses of both ...19921735186
antibodies to mycobacterium paratuberculosis in patients with crohn's disease.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 ...19911748044
lysis and fractionation of mycobacterium paratuberculosis and escherichia coli by matrix solid-phase dispersion.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 c ...19911750667
cultivation of mycobacterium paratuberculosis from bovine fecal specimens and a suggested standardized procedure. 19911760476
treatment of mycobacterium paratuberculosis infection in ruminants.paratuberculosis is a chronic, debilitating, fatal condition that usually is clinically undetectable until the onset of copious diarrhea. paratuberculosis is caused by an acid-fast organism, m. paratuberculosis. successful eradication of paratuberculosis depends on the early detection of infected animals, thereby allowing removal of carrier animals from the herd. treatment for paratuberculosis is therefore rarely indicated or undertaken; however, treatment may be considered for animals of except ...19911760762
mycobacterium paratuberculosis antigen d: characterization and evidence that it is a bacterioferritin.by using a combination of agarose and polyacrylamide gel electrophoresis, mycobacterium paratuberculosis antigen d was resolved from a crude sonicated preparation of the organism and characterized as a component with a molecular mass of approximately 400,000 da. while this component was composed mainly of protein, with unusually high proportions of glutamic acid and leucine, it was resistant to digestion with a number of proteolytic enzymes. structural detail revealed by electron microscopy, ami ...19911761687
chromatographic purification and characterization of antigens a and d from mycobacterium paratuberculosis and their use in enzyme-linked immunosorbent assays for diagnosis of paratuberculosis in sheep.the protein antigens a and d were purified from culture filtrates and sonic extracts of laboratory strains of mycobacterium paratuberculosis by salt precipitation and chromatography. the characterization of antigen a is shown here, and both antigens were evaluated along with lipoarabinomannan antigen in indirect enzyme-linked immunosorbent assays (elisa) for the serodiagnosis of ovine paratuberculosis. after anion-exchange (deae-5pw) and hydrophobic (phenyl-5pw) chromatography using high-perform ...19911761688
mass screening of cattle sera against 14 infectious disease agents, using an elisa system for monitoring health in livestock.mass screening elisa methods were developed for testing cattle serum for antibodies against 14 common livestock diseases simultaneously. the absorbance values were transformed to a %elisa (spectrophotometric antibody end point) by a computer interfaced with a microplate reader. a histogram indicating a cutoff point and a report for the veterinarian also was generated. the computer program produced a print-out of the antibody profile for each animal tested, the antibody concentration against each ...19911767993
associations between dairy production indices and lipoarabinomannan enzyme-immunoassay results for 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), wa ...19911790492
economic decision analysis model of a paratuberculosis test and cull program.a spreadsheet program was written to perform decision tree analysis for control of paratuberculosis (johne's disease), when testing all adults in a herd and culling all animals with positive test results. the program incorporated diagnostic test sensitivity, specificity, and test cost with the cost or value of each of the 4 possible outcomes; true-positive, true-negative, false-positive, and false-negative test results. the program was designed to repeat the analysis for the independent variable ...19911813465
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