Publications

relationships between fecal culture, elisa, and bulk tank milk test results for johne's disease in us dairy herds.	objectives were to estimate percentages of seropositive herds with cows shedding mycobacterium paratuberculosis in feces and milk, and to estimate sensitivity, specificity, and predictive value of an elisa relative to fecal culture. dairy cows (n = 712) were randomly selected from 61 herds previously identified by elisa as positive for johne's disease. fecal and bulk tank milk samples (n = 52 of 61 herds) were obtained from 10 states in the united states. fecal samples were processed by a double ...	2002	11949855
fibronectin attachment protein is necessary for efficient attachment and invasion of epithelial cells by mycobacterium avium subsp. paratuberculosis.	attachment and ingestion of mycobacterium avium subsp. paratuberculosis by two epithelial cell lines were enhanced by soluble fibronectin (fn). peptide blocking of the fn attachment protein (fap-p) inhibited the internalization of m. avium subsp. paratuberculosis. disruption of fap-p expression significantly reduced attachment and ingestion of m. avium subsp. paratuberculosis by t-24 and caco-2 cells. the results indicate that the interaction between fn and fap-p facilitates attachment and inter ...	2002	11953410
incidence of mycobacterium paratuberculosis in bulk raw and commercially pasteurized cows' milk from approved dairy processing establishments in the united kingdom.	over a 17-month period (march 1999 to july 2000), a total of 814 cows' milk samples, 244 bulk raw and 567 commercially pasteurized (228 whole, 179 semi-skim, and 160 skim), from 241 approved dairy processing establishments throughout the united kingdom were tested for the presence of mycobacterium paratuberculosis by immunomagnetic pcr (to detect all cells living and dead) and culture (to detect viable cells). overall, m. paratuberculosis dna was detected by immunomagnetic pcr in 19 (7.8%; 95% c ...	2002	11976118
molecular characterization of pigmented and nonpigmented isolates of mycobacterium avium subsp. paratuberculosis.	five pigmented isolates of mycobacterium avium subsp. paratuberculosis were examined by pulsed-field gel electrophoresis (pfge), is900 restriction fragment length polymorphism (is900-rflp), and is1311 polymorphism analysis using pcr. all of the pigmented isolates exhibited one of three distinct pfge profiles with snabi, designated 9, 10, and 11, and with spei, designated 7, 8, and 9, which generated three multiplex profiles designated [9-7], [10-8], and [11-9]. all of the pigmented isolates had ...	2002	11980962
synthesis and biological evaluation of new citrate-based siderophores as potential probes for the mechanism of iron uptake in mycobacteria.	several iron chelators containing alpha,beta-unsaturated hydroxamic acid motifs appended to a citric acid platform were synthesized. mycobacterium paratuberculosis was then challenged to grow in the presence of a panel of siderophores (mycobactin j, deferrioxamine b, acinetoferrin, and nannochelin a) and the new synthetic agents. of the structures tested, those containing the trans 2-octenoyl motif were preferred over those with trans cinnamoyl groups. in addition, derivatives containing longer ...	2002	11985473
the immunogenicity of mycobacterium paratuberculosis 85b antigen.	mycobacterium paratuberculosis (mpt) is the etiological agent of paratuberculosis. the disease is prevalent throughout the world, and exacts a heavy financial toll. at present, the only means of controlling this disease are culling or vaccination. the existing vaccines are not very efficient and produce a long-lasting local reaction at the point of injection and induce anti-bodies/delayed-type hypersensitivity (dth) reaction that cannot be differentiated from those of naturally infected animals. ...	2002	12005331
an is900-like sequence found in a mycobacterium sp. other than mycobacterium avium subsp. paratuberculosis.	the insertion sequence is900 has been considered specific for mycobacterium avium subsp. paratuberculosis (m. paratuberculosis) and has, therefore, been used as the target gene for diagnostic pcr of m. paratuberculosis. from a healthy dairy cow we have isolated and characterised a mycobacterium harbouring one copy of a sequence with 94% identity to is900 at the nucleic acid level. the isolate was shown to be related to mycobacterium cookii, as assessed by 16s rrna sequencing. strong amplificatio ...	2002	12007816
diagnosis of paratuberculosis in naturally infected goats.	a survey was carried out to verify if an immunohistochemical method associated with agar gel immunodiffusion (agid) will establish a firm diagnosis of caprine paratuberculosis. one hundred and thirty-six goats were tested by agid for antibodies against mycobacterium avium subsp. paratuberculosis at two different times: the first time 22 (19.1%) were positive and the second time 25 (18%). one seronegative goat with severe diarrhea and 5 seropositive goats, two of which showing similar clinical si ...	2002	12019718
detection methods of mycobacterium paratuberculosis.	mycobacterium paratuberculosis is receiving increasingly wider interest of scientific groups worldwide. this slow-growing mycobacterium does not only evoke paratuberculosis--an infectious cattle disease that brings huge economic losses--but it is also regarded as a potential cause of human crohn;s disease. it is very difficult to diagnose precisely this kind of animal infection in its very early stages, as well as to detect occurrence of m. paratuberculosis cells in the environment, including fo ...	2002	12448083
evaluation of two absorbed enzyme-linked immunosorbent assays and a complement fixation test as replacements for fecal culture in the detection of cows shedding mycobacterium avium subspecies paratuberculosis.	control of paratuberculosis in dairy herds is based on preventing the transmission of mycobacterium avium subsp. paratuberculosis (mptb) from cows to calves by management measures, supported by removal of cows excreting these bacteria by the fecal route (mptb shedders). fecal culture is the most accurate test for identifying mptb shedders, but this technique is expensive and takes up to 16 weeks for results to be available. serologic tests are inexpensive, rapid, and easy to perform. of serologi ...	2002	12033677
mycobacterium avium subsp. paratuberculosis strains from cattle and sheep can be distinguished by a pcr test based on a novel dna sequence difference.	a dna sequence differing between sheep and cattle types of mycobacterium avium subsp. paratuberculosis was identified and used to develop a pcr test. the test unequivocally distinguished all sheep types from cattle types and was negative for a wide range of other strains from the mycobacterium avium-mycobacterium intracellulare complex. the test will be useful for epidemiological purposes, particularly in hosts such as deer that can be easily infected with either type.	2002	12454189
analysis of mitogen-stimulated lymphocyte subset proliferation and nitric oxide production by peripheral blood mononuclear cells of captive elk (cervus elaphus).	elk (cervus claphus) are reservoirs for brucella abortus, mycobacterium bovis, and mycobacterium avium subsp. paratuberculosis, each a serious pathogen of domestic livestock. an understanding of the basic immune responsiveness of elk would aid efforts to develop methods to diagnose and prevent these diseases of elk. peripheral blood mononuclear cells (pbmc) isolated from captive elk were examined for phenotype, lymphocyte subset proliferative capacity, and ability to produce nitric oxide (no) up ...	2002	12038134
[establishment and evaluation of an elisa for the detection of antibodies in milk against mycobacterium avium subspecies paratuberculosis].	mycobacterium avium ssp. paratuberculosis (m. paratuberculosis) is the etiological agent of paratuberculosis (johne's disease), a chronic granulomatous enteritis of ruminants occurring worldwide with increasing frequency and leading to growing economic losses. continuous surveillance of dairy farms would be advisable, particularly with respect to the increasing economic importance of paratuberculosis and the high tenacity of the pathogen, which can persist in the environment for many months. so ...	2002	12073496
detection of mycobacterium avium subsp. paratuberculosis by buoyant density centrifugation, sequence capture pcr and dot blot hybridisation.	detection of mycobacterium avium subsp. paratuberculosis (m. paratuberculosis) by polymerase chain reaction (pcr) is often hampered by the lack of efficient methods for sample treatment. we report a protocol for analysis of faecal samples based on buoyant density centrifugation in percoll and is900 sequence capture pcr combined with a dot blot assay for detection of low-grade infection of m. paratuberculosis. serial dilutions of m. paratuberculosis genomic dna and m. paratuberculosis bacteria we ...	2002	12069770
mycobacterium paratuberculosis heat shock protein 70 as a tool in control of paratuberculosis.	paratuberculosis in cattle is a chronic intestinal disease in which a distinctive cellular reactivity of a th1-type preceeds the phase in which antibody titers are easily detectable and the animal becomes clinically ill. during infection with mycobacterium avium ssp. paratuberculosis (m.a.p), a decrease in cd4 t-helper cells has been observed in the clinical phase. our ultimate aim is to elicit a cytotoxic reaction against infected macrophages, using recombinant hsp70 (rhsp70) of m.a.p. as a too ...	2002	12072241
the mycobacterium avium subsp. paratuberculosis elisa response by parity and stage of lactation.	two cross-sectional studies were carried out to determine the enzyme-linked immunosorbent assay (elisa) response to mycobacterium avium subsp. paratuberculosis (map) by cow characteristics and stage of lactation. one of the studies (referred to as "milk-group") used milk samples from all lactating cows (n=7994) in 108 danish dairy herds. the other study (referred to as "serum-group") used serum samples collected from all cows (n=5323) in a subset of 72 herds from the 108 herds. these samples wer ...	2002	12062515
a fluorescence polarization assay for the detection of antibodies to mycobacterium bovis in cattle sera.	a fluorescence polarization assay (fpa) utilizing fluorescein-labelled mpb70 protein as the antigen was developed and evaluated for its ability to detect antibodies to mycobacterium bovis in cattle sera. three panels of sera were examined in this study. these included: (a) sera (n=28) obtained from cattle from which m. bovis was cultured; (b) sera (n=5666) from canadian field cattle which were presumed to be free from m. bovis; (c) sera (n=10) from cattle infected with mycobacterium paratubercul ...	2002	12034543
in situ identification of mycobacteria in crohn's disease patient tissue using confocal scanning laser microscopy.	the diversity in the methodology employed to investigate crohn's disease (cd) etiology has added significantly to the controversy of the mycobacterial role in this chronic inflammatory bowel disease. mycobacterium avium subsp paratuberculosis (map), a proposed and suspected agent in many cd patients, is a fastidious and very slow grower bacillus, which causes johne's disease (jd) in cattle. the methodology that has been widely and successfully used for isolation and identification of map from an ...	2002	12005446
infection and disease caused by environmental mycobacteria.	many species of mycobacteria that normally live as environmental saprophytes, the environmental mycobacteria (em), are opportunist causes of disease in humans and animals. many, but not all, cases are associated with some form of immune deficiency. an increasing number of species and clinical presentations are being described, and advances are being made in the understanding of the underlying predisposing factors. in recent years, four aspects of em disease have become particularly relevant to h ...	2002	11981303
application of the c(18)-carboxypropylbetaine specimen processing method to recovery of mycobacterium avium subsp. paratuberculosis from ruminant tissue specimens.	the causative agent of johne's disease is mycobacterium avium subsp. paratuberculosis. this is a chronic, debilitating gastrointestinal disorder that affects ruminants and is responsible for significant economic loss. the specimen processing method that combines c(18)-carboxypropylbetaine (cb-18) treatment and lytic enzyme decontamination has been shown to improve the diagnosis of mycobacterioses. this processing method was applied to the isolation of m. avium subsp. paratuberculosis from rumina ...	2002	11980960
open clinical trial of rifabutin and clarithromycin therapy in crohn's disease.	crohn's disease, an inflammatory bowel disease in humans, has a suspected aetiology of mycobacterium avium subsp. paratuberculosis.	2002	11930899
development and application of quantitative polymerase chain reaction assay based on the abi 7700 system (taqman) for detection and quantification of mycobacterium avium subsp. paratuberculosis.	numerous reports have described diagnostic methods based on the polymerase chain reaction (pcr) used to detect mycobacterium avium subsp. paratuberculosis, the causative agent of johne's disease. the result of conventional pcr tests has been only qualitative, either positive or negative; it does not present any quantitative information about the number of the agents in the specimen. a quantitative pcr method (is900 taqman) was developed to measure the number of m. a. paratuberculosis organisms p ...	2002	11939333
genome scale comparison of mycobacterium avium subsp. paratuberculosis with mycobacterium avium subsp. avium reveals potential diagnostic sequences.	the genetic similarity between mycobacterium avium subsp. paratuberculosis and other mycobacterial species has confounded the development of m. avium subsp. paratuberculosis-specific diagnostic reagents. random shotgun sequencing of the m. avium subsp. paratuberculosis genome in our laboratories has shown >98% sequence identity with mycobacterium avium subsp. avium in some regions. however, an in silico comparison of the largest annotated m. avium subsp. paratuberculosis contigs, totaling 2,658, ...	2002	11923349
treatment of severe crohn's disease using antimycobacterial triple therapy--approaching a cure?	mycobacterium avium subspecies paratuberculosis is probably the best candidate for a microbial cause of crohn's disease although arguments to the contrary can be equally convincing. growing evidence suggests that prolonged antimycobacterial combination therapy can improve crohn's disease in some patients.	2002	11926571
cell sorting of formalin-treated pathogenic mycobacterium paratuberculosis expressing gfp.	gfp is widely used as a molecular tool for the study of microbial pathogens. however, the manipulation of these pathogenic microorganisms poses a health threat to the laboratory worker, requiring biosafety level ii or iii containment. although the gfpfluorophore is tolerant toformalin, a thorough analysis of this treatment on fluorescent output in prokaryotic systems has not been described. in addition, the analysis of microorganisms expressing gfp often depends on specialized equipment, which m ...	2002	11911654
sensitivity and specificity of pooled faecal culture and serology as flock-screening tests for detection of ovine paratuberculosis in australia.	the flock-level sensitivity of pooled faecal culture and serological testing using agid for the detection of ovine johne's disease-infected flocks were estimated using non-gold-standard methods. the two tests were compared in an extensive field trial in 296 flocks in new south wales during 1998. in each flock, a sample of sheep was selected and tested for ovine johne's disease using both the agid and pooled faecal culture. the flock-specificity of pooled faecal culture also was estimated from re ...	2002	11849717
killing of mycobacterium avium subspecies paratuberculosis within macrophages.	mycobacterium avium subspecies paratuberculosis (m. paratuberculosis) is a facultative intracellular pathogen that resides within host macrophages during infection of ruminant animals. we examined survival of m. paratuberculosis infections within cultured macrophages to better understand the interplay between bacterium and host.	2002	11860602
crohn's disease caused by mycobacterium avium subspecies paratuberculosis: a public health tragedy whose resolution is long overdue.		2002	11800469
effect of commercial-scale high-temperature, short-time pasteurization on the viability of mycobacterium paratuberculosis in naturally infected cows' milk.	raw cows' milk naturally infected with mycobacterium paratuberculosis was pasteurized with an apv hxp commercial-scale pasteurizer (capacity 2,000 liters/h) on 12 separate occasions. on each processing occasion, milk was subjected to four different pasteurization treatments, viz., 73 degrees c for 15 s or 25 s with and without prior homogenization (2,500 lb/in(2) in two stages), in an apv manton gaulin kf6 homogenizer. raw and pasteurized milk samples were tested for m. paratuberculosis by immun ...	2002	11823197
antigen-induced production of interferon-gamma in samples of peripheral lymph nodes from sheep experimentally inoculated with mycobacterium avium subsp. paratuberculosis.	the production of interferon-gamma (ifn-gamma) in response to johnin purified protein derivate was measured in samples of the prescapular lymph node (pln) from 10 sheep, aged 2 years, and nine sheep, aged 1 year that had been inoculated orally with mycobacterium avium subsp. paratuberculosis within their first month of life. ten non-inoculated sheep, aged 1 year, constituted the negative control group. the results obtained in the pln ifn-gamma assay were compared with those derived from serologi ...	2002	11731176
quantitative analysis of mycobacterial and propionibacterial dna in lymph nodes of japanese and european patients with sarcoidosis.	the cause(s) of sarcoidosis is unknown. mycobacterium spp. are suspected in europe and propionibacterium spp. are suspected in japan. the present international collaboration evaluated the possible etiological links between sarcoidosis and the suspected bacterial species. formalin-fixed and paraffin-embedded sections of biopsy samples of lymph nodes, one from each of 108 patients with sarcoidosis and 65 patients with tuberculosis, together with 86 control samples, were collected from two institut ...	2002	11773116
comparison of real-time, quantitative pcr with molecular beacons to nested pcr and culture methods for detection of mycobacterium avium subsp. paratuberculosis in bovine fecal samples.	an automated pcr with fluorescent probes (molecular beacons) detected mycobacterium avium subsp. paratuberculosis in bovine feces. when the pcr was compared with culture in testing 41 fecal samples, kappa scores of 0.94 to 0.96, a sensitivity of 93 to 96%, and a specificity of 92% were obtained. results were quantitated by using a standard curve derived from a plasmid containing is900. a minimum quantity of 1.7 x 10(-4) pg of dna, correlating to 1 to 8 cfu, was detected.	2002	11773135
johne's disease in new zealand: the past, present and a glimpse into the future.	abstract extract • johne's disease is endemic in cattle and sheep in new zealand and is spreading in farmed deer. • procedures for the control of johne's disease in new zealand have been sporadic and often ineffective. • characterisation of strains of mycobacterium paratuberculosis using molecular biological techniques have revealed two distinct strain types. one type has been found in cattle, goats and deer, and the other in sheep, goats and deer • vaccination has been identified as an importan ...	2002	21838633
comparison between a commercial and an in-house elisa or anti-m. avium paratuberculosis antibodies detection in dairy herds in rio de janeiro, brazil.	paratuberculosis (johne's disease) is a chronic enteritis that affects ruminants and is caused by mycobacterium avium subsp. paratuberculosis (map). the disease is worldwide spread and causes important economic losses. in brazil, map has already been isolated, but there are no enough epidemiological studies about its prevalence. in this study, 179 sera of dairy cows originated from rio de janeiro state, brazil, were tested for the presence of anti-map antibodies using two elisas. first one was a ...	2002	17061486
mycobacterium avium subspecies paratuberculosis as a cause of crohn's disease.		2003	12477784
phagosomal maturation and intracellular survival of mycobacterium avium subspecies paratuberculosis in j774 cells.	the mechanisms by which mycobacterium avium subspecies paratuberculosis (m. a. ptb) survives within macrophages are not well characterized. one strategy for intracellular survival developed by mycobacterium tuberculosis is inhibition of phagosomal maturation. in this study it was hypothesized that m. a. ptb is capable of survival within macrophages by residing within a phagosomal compartment that does not mature into a functional phagolysosome. to test this hypothesis the following objectives we ...	2003	12676127
nymphs of the oriental cockroach (blatta orientalis) as passive vectors of causal agents of avian tuberculosis and paratuberculosis.	the potential transmission of the causal agent of paratuberculosis mycobacterium avium ssp. paratuberculosis and avian tuberculosis mycobacterium avium ssp. avium (actinomycetales: mycobacteriaceae) by nymphs of the oriental cockroach blatta orientalis l. (blattodea: blattidae) was investigated by oral infection with mycobacterial suspensions and examination of their droppings and bodies. both the subspecies of m. avium were isolated from droppings at 3 days post-infection and m. a. avium was fo ...	2003	12823831
is crohn's disease caused by a mycobacterium? comparisons with leprosy, tuberculosis, and johne's disease.	although crohn's disease is considered to be autoimmune in origin, there is increasing evidence that it may have an infectious cause. the most plausible candidate is mycobacterium avium subspecies paratuberculosis (map). intriguingly, koch's postulates may have been fulfilled for map and crohn's disease, even though they still have not been met for mycobacterium leprae and leprosy. in animals map causes johne's disease, a chronic wasting intestinal diarrhoeal disease evocative of crohn's disease ...	2003	12901893
mycobacterial interspersed repetitive units (miru) differentiate mycobacterium avium subspecies paratuberculosis from other species of the mycobacterium avium complex.	mycobacterial interspersed repetitive units (miru) comprise short tandem repeat structures found at multiple loci throughout the mycobacterium tuberculosis genome and have been used for typing these pathogens. we have identified miru at 18 conserved loci throughout the common portions of the mycobacterium avium subspecies paratuberculosis (map) and m. avium subspecies avium (maa) genomes. six of these loci were found to differ between maa and map in the number of tandem repeat motifs occurring a ...	2003	12944117
activation of macrophages and interference with cd4+ t-cell stimulation by mycobacterium avium subspecies paratuberculosis and mycobacterium avium subspecies avium.	mycobacterium avium subspecies paratuberculosis (m. ptb) and m. avium subspecies avium (m. avium) are closely related but exhibit significant differences in their interaction with the host immune system. the macrophage line, j774, was infected with m. ptb and m. avium and analysed for cytokine production and stimulatory capacity towards antigen-specific cd4+ t cells. under all conditions j774 cells were activated to produce proinflammatory cytokines. no influence on the expression of major histo ...	2003	12519304
development of the dutch johne's disease control program supported by a simulation model.	the development of a simulation model, "johnessim", was part of a research program aimed at designing a national johne's disease control program for the netherlands. initially, the focus was mainly directed towards different compulsory "test-and-cull" strategies. however, the results from the johnessim model showed that eradication of johne's disease based on such strategies would not be possible within 20 years and that it was also economically unattractive. however, improved calf management se ...	2003	12900150
an estimated prevalence of johne's disease in a subpopulation of alabama beef cattle.	the objective of this study was to estimate the overall prevalence of animals that were infected with mycobacterium avium ssp. paratuberculosis in a subpopulation of alabama beef cattle. this was determined using a commercial enzyme-linked immunosorbent assay (elisa) for the detection of m. avium ssp. paratuberculosis-specific antibodies in serum. serum was collected from 79 herds that were participating in the alabama brucellosis certification program. a total of 2,073 beef cattle were randomly ...	2003	12580290
application of different methods for the diagnosis of paratuberculosis in a dairy cattle herd in argentina.	paratuberculosis (ptbc) has a high prevalence in argentina, that affects dairy and beef cattle. the culture is the gold standard to the diagnosis of the disease. mycobacterium avium ssp. paratuberculosis (m. paratuberculosis), the aetiological agent, is difficult to isolate and grow in culture. in this study, 24 randomly selected cows of the fresian breed from a dairy herd with a history of ptbc were used to evaluate the performance of different diagnostic techniques. these animals did not show ...	2003	12710496
immunogenicity of a 16.7 kda mycobacterium paratuberculosis antigen.	mycobacterium paratuberculosis (mpt), the agent of paratuberculosis is a slow growing mycobacteria that causes important economic losses mainly due to lower weight gains and drastic decrease in milk production. existing paratuberculosis vaccines are not completely protective and induce antibodies/delayed type hypersensitivity (dth) reaction that cannot be differentiated from those of naturally infected animals. new potent acellular vaccines that allow discrimination between infected and vaccinat ...	2003	12623276
evaluation of serum and milk elisas for paratuberculosis in danish dairy cattle.	a milk and a serum elisa for detection of antibodies against mycobacterium avium ssp. paratuberculosis (map) were evaluated against the complement-fixation test (cft) and culture of faecal samples from 580 cows collected between august 1996 and december 1996. milk and serum were obtained concurrently from six dairy herds infected with map and from two dairy herds without history of infection with map.a cut-off value of 7 od% was used in the elisas. at this cut-off value, all six culture-positive ...	2003	12706056
evaluation of bacteriologic culture of individual and pooled fecal samples for detection of mycobacterium paratuberculosis in dairy cattle herds.	to determine the sensitivity of bacteriologic culture of pooled fecal samples in detecting mycobacterium paratuberculosis, compared with bacteriologic culture of individual fecal samples in dairy cattle herds.	2003	14552493
molecular epidemiology of mycobacterium avium subsp. paratuberculosis: evidence for limited strain diversity, strain sharing, and identification of unique targets for diagnosis.	the objectives of this study were to understand the molecular diversity of animal and human strains of mycobacterium avium subsp. paratuberculosis isolated in the united states and to identify m. avium subsp. paratuberculosis-specific diagnostic molecular markers to aid in disease detection, prevention, and control. multiplex pcr of is900 integration loci (mpil) and amplified fragment length polymorphism (aflp) analyses were used to fingerprint m. avium subsp. paratuberculosis isolates recovered ...	2003	12734243
pooled fecal culture sampling for mycobacterium avium subsp. paratuberculosis at different herd sizes and prevalence.	a stochastic spreadsheet model was developed to obtain estimates of the costs of whole herd testing on dairy farms for mycobacterium avium subsp. paratuberculosis (map) with pooled fecal samples. the optimal pool size was investigated for 2 scenarios, prevalence (a low-prevalence herd [< or = 5%] and a high-prevalence herd [> 5%]) and for different herd sizes (100-, 250-, 500- and 1,000-cow herds). all adult animals in the herd were sampled, and the samples of the individuals were divided into e ...	2003	12735345
is900/eric-pcr as a tool to distinguish mycobacterium avium subsp. paratuberculosis from closely related mycobacteria.	there is an increasing demand for fast and reliable methods to distinguish mycobacterium avium subsp. paratuberculosis (m. paratuberculosis) from closely related mycobacteria and also a need for rapid strain specific typing of clinical isolates for epidemiological reasons. in the present study, the potential of rep-pcr as a fingerprinting method for m. paratuberculosis was assessed and compared to conventional rflp. a pcr assay was designed and optimised to obtain reproducible fingerprints of my ...	2003	14559175
immune responses after oral inoculation of weanling bison or beef calves with a bison or cattle isolate of mycobacterium avium subsp. paratuberculosis.	paratuberculosis is endemic in domestic and wild ruminants worldwide. we designed the following study to compare host immune responses and pathologic changes in beef calves and bison calves after challenge with either a cattle or bison (bison bison) strain of mycobacterium avium subsp. paratuberculosis. in the first part of the study, six bison and six beef calves were orally inoculated with a cattle isolate of m. avium subsp. paratuberculosis over a 2 wk period. in the second part, an additiona ...	2003	14567215
a map to the diagnosis of crohn's disease.		2003	12769451
efficacy of commercial and field-strain mycobacterium paratuberculosis vaccinations with recombinant il-12 in a bovine experimental infection model.	the efficacy of commercial (strain 18) and field-isolate paratuberculosis vaccine preparations was investigated. the effect of prior exposure to mycobacterium paratuberculosis and the adjuvant effect of ril-12 on vaccine efficacy were also tested. both strain 18 and field-isolate vaccines induced strong local, systemic and enteric ifn-gamma responses. a significant reduction in mycobacterial colonization was observed when calves were vaccinated with the field-isolate prior to challenge, but not ...	2003	12804836
the results of using faecal culture as confirmation test of paratuberculosis-seropositive dairy cattle.	a total of 15,822 cattle aged 3 years and older, belonging to 378 randomly selected herds, were tested for paratuberculosis using an absorbed enzyme-linked immunosorbent assay (elisa); 3.3% tested positive. this percentage was lowest for the group of cattle aged 3-4 years (2.3%) and highest for cattle with the age of 5-6 years (4.5%). the mean sample to positive (s/p) ratio of seropositive cattle vaccinated against paratuberculosis was higher (0.75 +/- 0.33) than that of seropositive, non-vaccin ...	2003	12864898
diagnostic testing patterns of natural mycobacterium paratuberculosis infection in pygmy goats.	thirteen pygmy goats (capra hircus) from a herd naturally infected with mycobacterium avium ss. paratuberculosis (mptb) were monitored with 4 diagnostic assays for 2 to 15 mo. cellular and humoral immune responses to the infection were assessed with assays of gamma interferon (ifngamma), serum antibody [enzyme-linked immunosorbent assay (elisa) and agar gel diffusion (agid)], and radiometric fecal culture. microscopic examination and radiometric culture of tissue from 12 sites were performed at ...	2003	12889728
seroprevalence of johne's-disease infection in dairy cattle in california, usa.	a total of 1,950 serum samples from dairy cattle in california, usa were tested for the presence of antibodies to mycobacterium avium subspecies paratuberculosis (map) using a commercial enzyme-linked immunosorbent assay (elisa) kit. the sampled animals came from 65 herds and were sampled to reflect the relative numbers and distribution of dairy herds within three geographic regions in the state. using the manufacturers suggested cut-off for a positive test, 89 animals (4.6%) were positive. the ...	2003	12900163
analysis of repeated tests for interferon-gamma (ifn-gamma) response and faecal excretion for diagnosis of subclinical paratuberculosis in danish cattle.	a total of 315 cattle were tested for infection with mycobacterium avium subsp. paratuberculosis (m. paratuberculosis) at three consecutive samplings, using the interferon-gamma (ifn-gamma) test on whole blood and bacteriological culture of faecal samples. of 205 cattle from 10 infected herds 99 (48%) were positive in the ifn-gamma test on at least one sampling using "idexx-criteria" for interpretation, and of 110 cattle from five non-infected herds three (3%) were positive. forty-four animals f ...	2003	12909406
testing for mycobacterium avium subsp. paratuberculosis infection in asymptomatic free-ranging tule elk from an infected herd.	forty-five adult tule elk (cervus elaphus nannodes) in good physical condition were translocated from a population located at point reyes national seashore, marin county (california, usa), to a holding pen 6 mo prior to release in an unfenced region of the park. because infection with mycobacterium avium subsp. paratuberculosis (mptb) had been reported in the source population, the translocated elk underwent extensive ante-mortem testing using three johne's disease assays: enzyme linked immunoso ...	2003	12910759
early induction of humoral and cellular immune responses during experimental mycobacterium avium subsp. paratuberculosis infection of calves.	johne's disease (paratuberculosis) of cattle is widespread and causes significant economic losses for producers due to decreased production and poor health of affected animals. the chronic nature of the disease and the lack of a reproducible model of infection hinder research efforts. in the present study, instillation of mycobacterium avium subsp. paratuberculosis into the tonsillar crypts of neonatal calves resulted in peripheral colonization as detected by antemortem culture of feces and post ...	2003	12933856
preliminary characterization of chemically generated mycobacterium avium subsp. paratuberculosis cell wall deficient forms (spheroplasts).	cell wall deficient forms (cwd, spheroplasts) genetically indistinguishable from m. avium subsp. paratuberculosis (map) have been isolated from patients with crohn's disease and sarcoidosis. these map cwd may be important in the pathogenesis of these diseases and in johne's disease in other animal species. cwd forms are extremely difficult to isolate and generally revert to cell wall competent forms (cwc) when cultured in vitro. cultured map strain 19698 were chemically treated to generate suffi ...	2003	12935751
mycobacterium avium subsp. paratuberculosis infection causes suppression of rantes, monocyte chemoattractant protein 1, and tumor necrosis factor alpha expression in peripheral blood of experimentally infected cattle.	blood from cattle with subclinical mycobacterium avium subsp. paratuberculosis infection was stimulated with m. avium subsp. paratuberculosis antigens, and expression of interleukin-1beta (il-1beta), tumor necrosis factor alpha (tnf-alpha), rantes, monocyte chemoattractant protein 1 (mcp-1), and il-8 was measured. expression of tnf-alpha, rantes, and mcp-1 was lower in infected than in uninfected cattle. the reduced response may weaken protective immunity and perpetuate infection.	2003	14638822
use of loop-mediated isothermal amplification of the is900 sequence for rapid detection of cultured mycobacterium avium subsp. paratuberculosis.	we evaluated the usefulness of loop-mediated isothermal amplification (lamp) in detecting specific gene sequences of mycobacterium avium subsp. paratuberculosis (map). a total of 102 primer sets for lamp was designed to amplify the is900, hspx, and f57 gene sequences of map. using each of two primer sets (p-1 and p-2) derived from the is900 fragment, it was possible to detect map in a manner similar to that used with nested pcr. the sensitivity of lamp with p-1 was 0.5 pg/tube, which was more se ...	2003	12958269
evaluation of culture media for the recovery of mycobacterium avium subsp. paratuberculosis from cheddar cheese.	the study evaluated the efficacy of four mycobacterium avium subsp. paratuberculosis (map) culture media in suppressing commonly used starter cultures and typical nonstarter microflora present during the manufacture and ripening of cheddar cheese, with a view to identify a suitable medium for the enumeration of map during laboratory-scale cheddar production.	2003	12969490
lack of association of mycobacterium avium subsp. paratuberculosis with oocytes and embryos from moderate shedders of the pathogen.	paratuberculosis is a chronic and progressive disease of the intestine in ruminants caused by mycobacterium avium subsp. paratuberculosis (map). the bacterium is transmitted to young animals, becomes manifest in adulthood and leads to economic losses. the aim of this study is to investigate if cows shedding map possess oocytes and embryos that are carriers of the bacterium. new genetical material can enter the dairy farm using embryo transfer but the question as to whether this technique is safe ...	2003	12559469
the effect of decontamination protocols on the numbers of sheep strain mycobacterium avium subsp. paratuberculosis isolated from tissues and faeces.	the effect of decontamination protocols on the numbers of sheep strain mycobacterium avium subsp. paratuberculosis isolated in bactec cultures from clinical samples was assessed by spiking tissues and faeces at various points during the decontamination procedure. routine protocols in the laboratory were shown to decrease the number of organisms isolated per sample by about 2.7 log(10) and 3.1 log(10) for faeces and tissues, respectively. these findings are important for the interpretation of neg ...	2003	12935753
the mycobacterium avium subsp. paratuberculosis 35 kda protein plays a role in invasion of bovine epithelial cells.	mycobacterium avium subsp. paratuberculosis (m. paratuberculosis) enters intestinal epithelial cells of cattle and other ruminants via a mechanism that remains to be fully elucidated. this study showed that a gene encoding the m. paratuberculosis 35 kda major membrane protein (mmp) is expressed at a higher level in low-oxygen and high-osmolarity conditions that are similar to the environment of the intestine. in addition, cattle with johne's disease produced antibodies against mmp, suggesting th ...	2003	12904546
earthworms (oligochaeta, lumbricidae) and mycobacteria.	the objective of the study was to define the role of earthworms in the survival of mycobacteria in animal populations. in 13 sampling sites mycobacteria were detected in 53 (5.5%) samples of faeces and parenchymatous tissues from animals, in 25 (7.3%) environmental and in nine (8.2%) earthworm samples. in cattle and goat farms affected by mycobacterium avium subsp. paratuberculosis (m. paratuberculosis) of is900 restriction fragment length polymorphism (rflp) type b-c1 was isolated from 37 (4.6% ...	2003	12477646
genomic homogeneity between mycobacterium avium subsp. avium and mycobacterium avium subsp. paratuberculosis belies their divergent growth rates.	mycobacterium avium subspecies avium (m. avium) is frequently encountered in the environment, but also causes infections in animals and immunocompromised patients. in contrast, mycobacterium avium subspecies paratuberculosis (m. paratuberculosis) is a slow-growing organism that is the causative agent of johne's disease in cattle and chronic granulomatous infections in a variety of other ruminant hosts. yet we show that despite their divergent phenotypes and the diseases they present, the genomes ...	2003	12740027
detection by immunomagnetic pcr of mycobacterium avium subsp. paratuberculosis in milk from dairy goats in norway.	milk samples from 340 individual goats in 34 dairy herds throughout norway were examined for mycobacterium avium subsp. paratuberculosis (m.a. paratuberculosis) by culture and immunomagnetic separation combined with pcr (ims-pcr). the samples included three categories; (a) vaccinated dairy goats in herds with paratuberculosis; (b) vaccinated dairy goats in herds with no history of paratuberculosis; (c) unvaccinated goats in herds with no history of paratuberculosis. viable m.a. paratuberculosis ...	2003	12488077
comparative evaluation of the mgit and bactec culture systems for the recovery of mycobacterium avium subsp. paratuberculosis from milk.	aims: to compare the detection capabilities of the non-radiometric mgit (mycobacteria growth indicator tubes) and radiometric bactec 460tb culture systems (becton dickinson, cowley, oxford, uk) for recovering mycobacterium avium subsp. paratuberculosis from milk. methods and results: ultra heat treated (uht) milk samples spiked with different levels of m. paratuberculosis (10-107 cells ml-1) were inoculated into mgit and bactec media (containing recommended supplements) with and without prior ch ...	2003	12807471
detection and verification of mycobacterium avium subsp. paratuberculosis in fresh ileocolonic mucosal biopsy specimens from individuals with and without crohn's disease.	mycobacterium avium subsp. paratuberculosis is a robust and phenotypically versatile pathogen which causes chronic inflammation of the intestine in many species, including primates. m. avium subsp. paratuberculosis infection is widespread in domestic livestock and is present in retail pasteurized cows' milk in the united kingdom and, potentially, elsewhere. water supplies are also at risk. the involvement of m. avium subsp. paratuberculosis in crohn's disease (cd) in humans has been uncertain be ...	2003	12843021
adrenomedullin expression by gastric epithelial cells in response to infection.	many surface epithelial cells express adrenomedullin, a multifunctional peptide found in a wide number of body and cell systems. recently, we and others have proposed that adrenomedullin has an important novel role in host defense. this peptide has many properties in common with other cationic antimicrobial peptides, including the human beta-defensins. upon exposure of human gastric epithelial cells to viable cells of invasive or noninvasive strains of helicobacter pylori, escherichia coli, salm ...	2003	12853384
use of growth indices from radiometric culture for quantification of sheep strains of mycobacterium avium subsp. paratuberculosis.	a simple method for using growth indices from radiometric bactec cultures was evaluated for the enumeration of australian sheep strains of mycobacterium avium subsp. paratuberculosis. the numbers of viable organisms in inocula were determined by end-point titration in bactec cultures. growth indices were measured by using a bactec 460 machine. there was a linear relationship between the number of days taken for the cumulative growth index to reach 1,000 (dcgi1000) and log(10) inoculum size. the ...	2003	12788757
comparison of two dna extractions and nested pcr, real-time pcr, a new commercial pcr assay, and bacterial culture for detection of mycobacterium avium subsp. paratuberculosis in bovine feces.	in this study, 5 combinations of 2 dna extractions and 3 polymerase chain reaction (pcr) techniques were compared with culture for the detection of mycobacterium paratuberculosis directly from bovine feces. these combinations included a new commercial extraction technique combined with a commercial pcr/southern blot technique, nested pcr (npcr), or real-time pcr, and a university-developed extraction combined with npcr or real-time pcr. four of the 5 combinations had statistically similar sensit ...	2003	12661717
temporal patterns of diagnostic results in serial samples from cattle with advanced paratuberculosis infections.	as part of investigating diagnostic strategies for mycobacterium avium subsp. paratuberculosis (map), serial results from polymerase chain reaction (pcr) on extraintestinal tissues (blood, milk, and liver) were compared with those from more conventional detection methods including serum enzyme-linked immunosorbent assay (elisa), fecal culture, and fecal pcr. three cows previously identified as being subclinically infected with map were selected for the study. blood, milk, and feces were collecte ...	2003	12661735
unanswered questions about the transmission of mycobacterium avium subspecies paratuberculosis.		2003	12672360
use of bioinformatics to predict a function for the gs element in mycobacterium avium subspecies paratuberculosis.	mycobacterium avium subsp. paratuberculosis (map) is a member of the mycobacterium avium complex (mac) and causes the inflammatory bowel disease, johne's disease, in livestock. map has also been implicated as the causative agent of a similar disease, crohn's disease, in humans. one of three major genetic differences between map and non-pathogenic mac is the 6496-bp gs element. based on the output from freely available protein sequence and structural bioinformatics tools, and the close homology o ...	2003	12673062
mycobacterium paratuberculosis in dairy herds in alberta.	fifty dairy herds in alberta were tested for the presence of mycobacterium paratuberculosis by fecal culture and serum enzyme linked immunosorbent assay (elisa). individual sera (1500) were tested for antibodies to m. paratuberculosis by elisa. fecal samples were combined in pools of 3 (10 pools/herd) for a total of 500 pools that were cultured for m. paratuberculosis. thirty cultures, including all 10 pools from 1 herd, were not readable due to fungal contamination. the remaining 470 cultures, ...	2003	12677691
the use of spiral plating and microscopic colony counting for the rapid quantitation of mycobacterium paratuberculosis.	to evaluate a spiral plating and microscopic colony counting technique to hasten the quantitation of mycobacterium paratuberculosis.	2003	12680941
do non-ruminant wildlife pose a risk of paratuberculosis to domestic livestock and vice versa in scotland?	paratuberculosis (johne's disease) was long considered only a disease of ruminants. recently non-ruminant wildlife species have been shown to harbor mycobacterium avium subsp. paratuberculosis, the causative organism of paratuberculosis. we review the known non-ruminant wildlife host range of m. avium subsp. paratuberculosis and consider their role in the epidemiology of paratuberculosis in domestic ruminant livestock. mycobacterium avium subsp. paratuberculosis has been isolated from lagomorph, ...	2003	12685064
longitudinal study to investigate variation in results of repeated elisa and culture of fecal samples for mycobacterium avium subsp paratuberculosis in commercial dairy herds.	to determine sources and amounts of variation in a kinetics elisa (kela) and results of culture of fecal samples for mycobacterium avium subsp paratuberculosis (map) in repeated tests of individual cows.	2003	12693540
the emergence of mycobacterium paratuberculosis in farmed deer in new zealand - a review of 619 cases.	to review cases in which mycobacterium paratuberculosis was identified in farmed deer in new zealand.	2003	16032301
absence of mycobacterium avium subspecies paratuberculosis components from crohn's disease intestinal biopsy tissues.	crohn's disease is a chronic human intestinal inflammatory disorder for which an etiologic agent has not been identified. johne's disease is a similar chronic enteric granulomatous disease of ruminant species and has been used as a model of crohn's disease. johne's disease has been proven to be caused by mycobacterium avium subspecies paratuberculosis (m. avium ss paratuberculosis). it has been proposed that m. avium ss paratuberculosis may also cause crohn's disease. this is of particular conce ...	2003	15931311
targeting enteric bacteria in treatment of inflammatory bowel diseases: why, how, and when.	this review discusses the role of bacterial adjuvants and antigens in induction and reactivation of chronic intestinal inflammation in susceptible hosts; discusses the results of recent therapeutic trials of antibiotics, probiotics, and prebiotics; and suggests future treatment strategies.	2003	15703577
elevated antibody responses in patients with crohn's disease against mpp14, a 14 kda secreted protein purified from mycobacterium avium subsp. paratuberculosis.		2003	15074652
mycobacterium avium subspecies paratuberculosis--a review of present research in norway.		2003	15074645
ongoing research on paratuberculosis in sweden.		2003	15074647
factors affecting the herd level of antibodies against mycobacterium avium subsp. paratuberculosis--a case-control study.		2003	15074650
culture of pooled tissues for the detection of mycobacterium avium subsp paratuberculosis infection in individual sheep.		2003	15080490
evaluation of two recovery methods for detection of mycobacterium avium subsp. paratuberculosis by pcr: direct-dilution--centrifugation and c(18)-carboxypropylbetaine processing.	a duplex polymerase chain reaction (pcr)-hybridization assay based on mycobacterium avium subsp. paratuberculosis (map)-specific is900 integration sites was used to evaluate two mycobacterial recovery methods from bovine feces: a direct-dilution-centrifugation method and a c(18)-carboxypropylbetaine (cb-18)-based method. all map pcr results were confirmed for absence of inhibitors using a novel pcr system based on the rpob gene of plant chloroplasts as an internal control. the detection limits o ...	2003	14680691
comparison of histopathology, cultivation of tissues and rectal contents, and interferon-gamma and serum antibody responses for the diagnosis of bovine paratuberculosis.	the diagnosis of infection with mycobacterium avium subsp. paratuberculosis was studied in 12 cattle from infected dairy herds and four from non-infected herds. a comparison was made of (1) histopathological examination and bacterial culture of tissues, (2) culture of serial samples of rectal contents, and (3) examination of repeated blood samples for interferon-gamma (ifn-gamma) and antibody responses. tissue samples were taken from the small and large intestine and corresponding mesenteric lym ...	2003	14554124
experimental infection of weaner sheep with s strain mycobacterium avium subsp. paratuberculosis.	we sought to determine whether infection of recently weaned 12-16-week-old merino lambs with an australian s strain m. a. paratuberculosis, at doses consistent with natural exposure, could be detected in the first few months post-inoculation. such detection would facilitate the use of weaner sheep as sentinel animals for the presence of infectious doses of m. a. paratuberculosis on pastures. in controlled pen trials, oral doses of approximately 10(7)-10(8) viable organisms were demonstrated to b ...	2003	14559172
enhanced expression of interleukin-1alpha and tumor necrosis factor receptor-associated protein 1 in ileal tissues of cattle infected with mycobacterium avium subsp. paratuberculosis.	infection with mycobacterium avium subsp. paratuberculosis is associated with high levels of morbidity, decreased production, and early culling in dairy cattle. clinical symptoms of johne's disease include persistent diarrhea, inappetence, and resultant weight loss due to chronic inflammation of the small intestine. although the presence or absence of intestinal lesions cannot be used as a definitive indicator of m. avium subsp. paratuberculosis infection, most infected cattle exhibit significan ...	2003	14573670
evidence for a novel gene expression program in peripheral blood mononuclear cells from mycobacterium avium subsp. paratuberculosis-infected cattle.	a bovine-specific cdna microarray system was used to compare gene expression profiles of peripheral blood mononuclear cells (pbmcs) from control uninfected (n = 4) and johne's disease-positive (n = 6) holstein cows. microarray experiments were designed so that for each animal, a direct comparison was made between pbmcs stimulated in vitro with mycobacterium avium subsp. paratuberculosis and pbmcs stimulated with phosphate-buffered saline (nil-stimulated pbmcs). as expected, m. avium subsp. parat ...	2003	14573671
variation in resistance of mycobacterium paratuberculosis to acid environments as a function of culture medium.	acid resistance of mycobacterium paratuberculosis was examined as a function of growth conditions (i.e., in vitro growth medium and ph). m. paratuberculosis was cultured in either fatty acid-containing medium (7h9-oadc) or glycerol-containing medium (wr-gd or 7h9-gd) at two culture phs (phs 6.0 and 6.8). organisms produced in these six medium and ph conditions were then tested for resistance to acetate buffer at phs 3, 4, 5, and 6 at 20 degrees c. a radiometric culture method (bactec) was used t ...	2003	14602647
study of animal-borne infections in the mucosas of patients with inflammatory bowel disease and population-based controls.	crohn's disease may be triggered by an infection, and it is plausible to consider that such an infection may be animal borne and ingested with our food. there has been considerable interest in the past in determining whether mycobacterium avium subsp. paratuberculosis (m. avium) might be the etiologic agent in crohn's disease since it causes a disease in cattle that is similar to crohn's disease in humans. we aimed to determine if there was an association between crohn's disease and infection wi ...	2003	14605128
prevalence estimates for paratuberculosis adjusted for test variability using bayesian analysis.	the elisa tests that are available to detect an infection with mycobacterium avium subsp. paratuberculosis (map) have a limited validity expressed as the sensitivity (se) and specificity (sp). in many studies, the se and sp of the tests are treated as constants and this will result in an underestimation of the variability of the true prevalence (tp). bayesian inference provided a natural framework for using information on the test variability (i.e., the uncertainty) in the estimates of test se a ...	2003	12941553
[mycobacterium avium subspecies paratuberculosis].		2003	14716909
characterization of genetic differences between mycobacterium avium subsp. paratuberculosis type i and type ii isolates.	a combination of representational difference analysis and comparative dna sequencing revealed that four type i (sheep) isolates of mycobacterium avium subsp. paratuberculosis were differentiated from nine type ii (bovine) isolates by the presence of an 11-bp insertion in a novel m. avium subsp. paratuberculosis-specific region of genomic dna. further, our studies show that m. avium subsp. paratuberculosis type i isolates contain three type-specific loci that are missing in m. avium subsp. paratu ...	2003	14605167
evaluation of an absorbed elisa and an agar-gel immuno-diffusion test for ovine paratuberculosis in sheep in australia.	the sensitivities and specificities of an absorbed enzyme-linked immunosorbent assay (elisa) and an agar-gel immuno-diffusion (agid) test for the detection of johne's disease in sheep were estimated using data from six known infected and 12 assumed uninfected sheep flocks. sensitivities were estimated for all histologically positive sheep, as well as by histological lesion score, lesion type (paucibacillary or multibacillary) and sheep body-condition score, with elisa sensitivities estimated at ...	2003	14623409
b-cell epitopes in the immunodominant p34 antigen of mycobacterium avium ssp. paratuberculosis recognized by antibodies from infected cattle.	mycobacterium avium ssp. paratuberculosis (m. paratuberculosis) causes johne's disease, a chronic and fatal enteritis in ruminants. in the last stage of the disease, antibody titres rise and levels of interferon-gamma decrease, suggesting that the host-immune response is switching from a t helper 1 (th1) to a th2 profile. in infected cattle, the membrane protein p34 elicits the predominant humoral response against m. paratuberculosis. to map the b-cell epitopes of this antigen, affinity-purified ...	2003	14629623