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gen-probe rapid diagnostic system for distinguishing between mycobacterium avium and mycobacterium paratuberculosis. 19911719027
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
mycobacterium paratuberculosis and crohn's disease.the possible aetiological role of mycobacterium paratuberculosis in crohn's disease was investigated. the immunological response was studied using an enzyme-linked immunosorbent assay (elisa), western blotting, and immunocytochemistry. the antibody response to two protoplasmic antigen preparations of m paratuberculosis in the sera of patients with inflammatory bowel disease was measured by elisa. igg and igm antibodies to these antigens were measured in serum samples from 52 patients with crohn' ...19911991637
crossed immunoelectrophoretic analysis of mycobacterium paratuberculosis.antigenic analysis of m. paratuberculosis revealed extensive cross-reactivity with m. avium; however, the number of cross-reactive antigens found was dependent on the strain of m. avium tested. one antigen was shown to be the common antigen while another appeared to be iron-regulated in its production. a commercial polyclonal antibody to m. paratuberculosis produced a cie precipitin pattern comparable to that of the antibody produced for the present study. an antigen designated no. 6 was consist ...19911993120
evaluation of a commercial enzyme-linked immunosorbent assay for johne's disease.a new commercial kit for diagnosis of bovine paratuberculosis (johne's disease), called the johne's absorbed eia (enzyme immunoassay; commonwealth serum laboratories, parkville, victoria, australia), was evaluated by using serum specimens from the national repository for paratuberculosis specimens. the evaluation was specifically designed to measure test sensitivity and specificity for detection of dairy cattle with subclinical paratuberculosis. the case definition of subclinical bovine paratube ...19912007634
serodiagnosis of bovine paratuberculosis by use of a dot enzyme-linked immunosorbent assay.a dot elisa was developed for detection of antibodies to mycobacterium paratuberculosis. the assay was evaluated by testing sera from cattle that were determined, by bacteriologic culturing of feces, to be infected with m paratuberculosis and were suspected of having clinical disease. further evaluation involved testing sera from cattle in which m paratuberculosis had not been isolated from feces on several attempts. results of the dot elisa were positive for sera from 86 of 101 infected cattle, ...19912012331
a dna probe for the detection of mycobacterium paratuberculosis.a genomic library of dna extracted from mycobacterium paratuberculosis was constructed in the expression vector lambda gt 11. the library was screened by plaque hybridization with labelled m. paratuberculosis genomic dna as probe. strongly hybridizing plaques were isolated and their dna extracted and characterised for m. paratuberculosis specificity by hybridization to dna from other mycobacteriaceae. a clone was obtained which was specific for m. paratuberculosis. dna from this clone could dete ...19912024441
gen-probe rapid diagnostic system for the mycobacterium avium complex does not distinguish between mycobacterium avium and mycobacterium paratuberculosis.three reference and 16 field strains of mycobacterium paratuberculosis were tested with the gen-probe mycobacterium avium complex dna probe (gen-probe inc., san diego, calif.). all reference strains and 12 of 16 field strains gave positive hybridization results with the probe. this study shows that the m. avium complex probe does not distinguish between m. avium and m. paratuberculosis and indicates heterogeneity in the 16s rrna gene of m. paratuberculosis.19912037681
antibody reactivities of mycobacterium paratuberculosis infected sheep as analyzed by enzyme-linked immunosorbent assay and western blotting.antibody reactivities in sera from mycobacterium paratuberculosis (m. ptb) infected and vaccinated sheep were analyzed by enzyme-linked immunosorbent assay (elisa) and western (immuno)blotting using a sonicate antigen from m. ptb. both methods allowed good differentiation between infected/vaccinated animals and noninfected controls. removal of nonspecific crossreactive antibodies by absorption with a m. phlei sonicate antigen coupled to sepharose reduced elisa reactivities of positive sera by 50 ...19912040422
mycobacterium paratuberculosis monoassociated nude mice as a paratuberculosis model.in this study, a paratuberculosis (johne's disease) model was developed by intragastrically dosing gnotobiotic athymic nude mice with mycobacterium paratuberculosis. the mice infrequently shed bacilli from their intestinal tracts during the first 4 months after inoculation. following this time, increasing numbers of m. paratuberculosis (greater than 4.0 log10 bacilli per fecal pellet by 40 weeks) were recovered from the feces of the 12 mice that remained in the isolator. a similar pattern of rec ...19912063515
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
induction of suppressor cells by mycobacterium paratuberculosis antigen in inflammatory bowel disease.we studied the m. paratuberculosis-induced proliferation and suppressor cell generation by peripheral blood lymphocytes from patients with inflammatory bowel disease. peripheral blood lymphocytes were separated from 33 patients with crohn's disease, 18 with ulcerative colitis, nine with other intestinal diseases, and five with autoimmune disorders. proliferation of peripheral blood lymphocytes from normal individuals in response to 10 micrograms/ml of m. paratuberculosis antigen was reduced by d ...19911825189
experimental disease in young chickens induced by a mycobacterium paratuberculosis isolate from a patient with crohn's disease.the susceptibility of young chickens to infection with an isolate of mycobacterium paratuberculosis which had been recovered from diseased tissues of a patient with crohn's disease was determined. two-week-old leghorn-cochin chicks were inoculated with strain linda. six birds received 10(7) organisms orally, six intraperitoneally, and five intracardially. six uninoculated birds served as contact controls. birds from each group were necropsied at two-week intervals. focal granulomatous lesions oc ...19911832080
evaluation of enzyme-linked immunosorbent assay in comparison with complement fixation test for the diagnosis of subclinical paratuberculosis in cattle.an enzyme-linked immunosorbent assay (elisa) was evaluated and compared in parallel with the standard complement fixation test (cft) for the diagnosis of bovine subclinical paratuberculosis. bovine sera preabsorbed with the mixture of mycobacterium phlei and kaolin suspension were assayed for antibody activities to the crude protoplasmic antigen of mycobacterium paratuberculosis in the elisa. elisa antibody titer was expressed as elisa antibody index (eai) value: eai = (at-an)/(ap-an), where at, ...19911834198
lectinhistochemical study on normal and paratuberculosis-affected bovine ileum. 19911834225
correction of antigen-specific t cell defects in aged murine gut-associated lymphoid tissues an immune intervention by combined adoptive transfer of an antigen-specific immunoregulatory cd4 t cell subset and interleukin 2 administration.gut-associated lymphoid tissues (galt) from aged mice enterically immunized with mycobacterium paratuberculosis protoplasmic antigen show hyperreactive humoral immune responses; this hyperresponsiveness can be corrected to a considerable extent, but not entirely, by systemic administration of interleukin 2 (il 2) alone. the aim of the present study was to determine further whether the hyperreactivity in the antigen-specific humoral immune responses in aged galt could be fully restored by adoptiv ...19911836186
suppressive effect of nonviable mycobacterium paratuberculosis on the delayed-type hypersensitivity reaction to sheep erythrocytes in mice.the effect of nonviable mycobacterium paratuberculosis on the delayed-type hypersensitivity reaction to sheep erythrocytes (srbc) in mice was evaluated by means of delayed-type footpad swelling. intraperitoneal (i.p.) injection with nonviable m. paratuberculosis into mice from 28 days before to 1 day after immunization with srbc resulted in a significant suppression of foot-pad swelling to srbc. the suppressive effect could be transferred by i.p. injection of spleen cells or peritoneal exudate c ...19911836513
specific detection of mycobacterium paratuberculosis by dna hybridisation with a fragment of the insertion element is900.this paper describes the evaluation of a newly developed dna probe for mycobacterium paratuberculosis. dna probe pcr278 is a 278 bp fragment obtained by polymerase chain reaction (pcr) amplification of the 5'-region of is900, an insertion element contained in the genome of m paratuberculosis. this dna probe can specifically distinguish m paratuberculosis from a wide range of other organisms, including members of the m avium-m intracellulare complex. when used in conjunction with the pcr amplific ...19911851124
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 ...19911883088
development and evaluation of a rapid absorbed enzyme immunoassay test for the diagnosis of johne's disease in cattle.an absorbed enzyme immunoassay (eia) test for johne's disease in cattle was developed in which absorption of cross-reacting antibodies occurred as a rapid reaction in solution rather than overnight with whole organisms and a subsequent centrifugation step. total test time was reduced to less than 2 h with a minimum of manipulations. the test was evaluated in cattle herds from johne's disease-endemic and johne's disease-free regions of australia. specificity was 99.8%. calculations of sensitivity ...19911883290
complete nucleotide sequence of a gene encoding the 70 kd heat shock protein of mycobacterium paratuberculosis. 19911886778
an epidemiological study of paratuberculosis in dairy cattle in ontario: study design and prevalence estimates.an observational study involving 304 dairy herds and three abattoirs was conducted between 1986 and 1989 to investigate the epidemiology of paratuberculosis in dairy cattle in ontario. the objectives of this paper were to describe the method of data collection for the study, to present descriptive production statistics and to present estimates of the prevalence of paratuberculosis among dairy cattle in ontario. mycobacterium paratuberculosis was isolated from the distal ileum and/or the ileoceca ...19911889036
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 ...19911909601
reactivity of fungal organisms in tissue sections using anti-mycobacteria antibodies.sixty-four cases of deep fungal infections diagnosed using pas or silver stains and 18 control cases of sarcoidosis, m. tuberculosis and m. leprae infection were stained using commercial polyclonal antibody raised against m. paratuberculosis (mp), m. duvalii (md), and bacillus calmette-guerin (bcg). nine of 13 cases of sporotrichosis stained positively using anti-mp antibody only; 13 of 14 cases of histoplasmosis stained with anti-bcg, anti-md, and anti-mp; seven cases of cryptococcosis had only ...19911918507
serologic enzyme-linked immunosorbent assay responses of calves vaccinated with a killed mycobacterium paratuberculosis vaccine.the purpose of this study was to document the effect of calfhood vaccination for mycobacterium paratuberculosis on a serologic elisa. fifteen calves vaccinated with a killed paratuberculosis vaccine and 5 unvaccinated control calves were tested from the first through the fifteenth month of life. age of vaccination ranged from 5 to 40 days. blood samples were collected prior to vaccination and periodically thereafter. serum antibody was analyzed by use of the elisa. all calves were elisa-negative ...19911928899
comparison of different media for the isolation of small ruminant strains of mycobacterium paratuberculosis.two different egg based media, with and without the incorporation of sodium pyruvate, were used to isolate m. paratuberculosis from sheep, goat and cattle samples obtained at our laboratory for two years. both media were adequate for bovine material, with a slightly improved isolation rate for herrold's egg yolk medium incorporating sodium pyruvate; however, most of the small ruminant strains grew only on löwenstein-jensen medium without sodium pyruvate. those results point out the need to use d ...19911949552
spontaneous murine thymocyte comitogenic activity consistent with interleukin-1 in cattle naturally infected with mycobacterium paratuberculosis.the production of comitogenic activity consistent with interleukin-1 (il-1) activity by blood monocytes from cattle with naturally acquired paratuberculosis was examined by murine thymocyte proliferation. in addition, il-1-like activity in response to homologous and heterologous antigens was determined. activity was determined in nine cattle naturally infected with mycobacterium paratuberculosis and six non-infected cattle. comitogenic properties were measured in response to m. paratuberculosis ...19911949571
comparison of the johne's absorbed eia and the complement-fixation test for the diagnosis of johne's disease in cattle.a commercially available absorbed elisa for the diagnosis of johne's disease (jd) (paratuberculosis) in cattle, the johne's absorbed eia, was compared with the conventional complement-fixation test (cft) used in australia. stored plasma from 3 victorian dairy herds with a history of jd, sera from specimens submitted from animals showing clinical signs of jd and sera from the us national repository for paratuberculosis specimens were used to determine the sensitivity of each test. the eia detecte ...19911953547
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
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
cloning and expression of portions of the 34-kilodalton-protein gene of mycobacterium paratuberculosis: its application to serological analysis of johne's disease.paratuberculosis (johne's disease), an endemic mycobacteriosis of cattle that is caused by mycobacterium paratuberculosis, is characterized by incoercible diarrhea and fecal shedding of bacteria. the present work aimed at developing a specific serological test for this disease. we have recently shown that a 34-kda protein belonging to the major antigen complex a36 of m. paratuberculosis is immunodominant and contains epitopes specific with respect to all mycobacteria tested, including mycobacter ...19937681851
rapid detection and identification of mycobacterium avium by amplification of 16s rrna sequences.an assay that is based on the amplification of 16s rrna sequences and that was initially developed to detect mycobacterium paratuberculosis in cattle was used to test 20 serotypes of the mycobacterium avium complex (mac) and atypical mycobacterial species not belonging to mac. only serotypes 1 to 6 and 8 to 11, designated m. avium, were detected by the assay, indicating that it can be used for the rapid detection and identification of m. avium. the results of the assay for clinical samples from ...19937691877
[antigen-binding lymphocytes to the mycobacterium paratuberculosis antigen in crohn's disease and nonspecific ulcerative colitis]. 19937521551
identification of mycobacteria from animals by restriction enzyme analysis and direct dna cycle sequencing of polymerase chain reaction-amplified 16s rrna gene sequences.two methods, based on analysis of the polymerase chain reaction-amplified 16s rrna gene by restriction enzyme analysis (rea) or direct cycle sequencing, were developed for rapid identification of mycobacteria isolated from animals and were compared to traditional phenotypic typing. bactec 7h12 cultures of the specimens were examined for "cording," and specific polymerase chain reaction amplification was performed to identify the presence of tubercle complex mycobacteria. combined results of sepa ...19937508456
comparison of polymerase chain reaction tests and faecal culture for detecting mycobacterium paratuberculosis in bovine faeces.a polymerase chain reaction (pcr) test for m. paratuberculosis was developed based on a 218 bp segment of a dna insertion sequence, is900, that is specific for this organism. the method involved two consecutive amplification reactions, with the second set of primers being nested inside the first set. the method reliably detected 50 organisms/g faeces. this pcr test was applied to 32 bovine faecal specimens containing high, moderate or low numbers of m. paratuberculosis organisms as determined by ...19937794290
investigation of mycobacterium paratuberculosis in sheep by faecal culture, dna characterisation and the polymerase chain reaction. 19938116172
abolish mycobacterium paratuberculosis strain 18. 19938123098
serologic reactivity against mycobacterium paratuberculosis antigens in patients with sarcoidosis.although sarcoidosis has clinical and histopathologic similarities to some forms of tuberculosis and other mycobacterial infections, attempts to establish a mycobacterial etiology have not been successful. using cytoplasmic antigens derived from a wild strain of mycobacterium paratuberculosis in an enzyme-linked immunosorbent assay, patients with sarcoidosis were found to have immunoglobulin levels significantly higher than those found in a control population in the igg, but not in iga or igm an ...19938134712
use of rifabutin in treatment of systemic mycobacterium paratuberculosis infection in mice.balb/c mice were infected intraperitoneally with mycobacterium paratuberculosis and, after allowing the infection to progress for 30 days, were treated with rifabutin at 0, 12.5, 25, and 50 mg/kg of body weight. rifabutin was administered in drinking water under conditions of water deprivation, whereby the entire daily dose was delivered within a 1-h period. animals were killed at biweekly intervals from time zero of treatment to 180 days. spleens and livers from each animal were examined by qua ...19938215277
the dangers of disease transmission by artificial insemination and embryo transfer.this review summarizes the major infectious diseases of the three major agricultural species (cattle, sheep and pigs) and horses, and presents the evidence for and against the possibility of infectious agents being transmitted between animals via the venereal route or by the use of semen or early embryos in commercial artificial insemination (ai) or embryo transfer (et). cattle feature most prominently in the widespread distribution of frozen semen, and national and international organizations h ...19938221041
control of johne's disease in a flock of sheep by vaccination. 19938236728
isolation of mycobacterium paratuberculosis from mononuclear cells in tissues, blood, and mammary glands of cows with advanced paratuberculosis.seven mature dairy cows from 6 herds were obtained with history, clinical signs of disease, and laboratory findings suggestive of advanced paratuberculosis. a surgically implanted collection chamber was used to obtain peripheral tissue fluid. blood, mammary gland flush fluid, and collection chamber flush fluid (ccff) samples were obtained 6 times over a 2-week period from each cow. mononuclear cell-rich portions of these fluids obtained by gradient centrifugation were submitted for bacteriologic ...19938239130
accuracy and economics of the usda-licensed enzyme-linked immunosorbent assay for bovine paratuberculosis. 19938276710
search for mycobacterium paratuberculosis dna in tissue from patients with sarcoidosis by enzymatic gene amplification.the etiology of sarcoidosis has not yet been established, but several mycobacterial species have been implicated in the pathogenesis of this disease. we have used a nested primer polymerase chain reaction (pcr) specific for the is900 mycobacterium paratuberculosis insertion element to analyse paraffin-embedded mediastinal lymphoid tissue from 18 patients with pulmonary sarcoidosis verified by mediastinoscopy. only the positive control contained m. paratuberculosis-specific dna. the present study ...19938286096
paratuberculosis in farmed deer: case reports and dna characterization of isolates of mycobacterium paratuberculosis.mycobacterium paratuberculosis was isolated from farmed deer in new zealand on 21 occasions over a 7-year period. the number of cases increased during the last 3 years of the study. clinical paratuberculosis was observed in 5 deer, 3 other cases came from grossly normal animals that reacted to a tuberculin skin test, and the remaining 13 animals had tuberculous lesions that were identified at meat inspection. pathologic features of the lesions in these 13 cases included necrosis and mineralizati ...19938286456
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