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PubMed2004

An evaluation of mycophage therapy, chemotherapy and vaccination for control of Mycobacterium avium subsp. paratuberculosis infection.

The control of ovine Johne's disease (OJD) is important for domestic trade, the viability of farming units and possibly also for public health. Current strategies in Australia have included quarantine and pasture spelling to decrease prevalence and shedding rates and reduce numbers of Mycobacterium paratuberculosis (Mptb) ingested by susceptible sheep. However, alternative procedures are needed and vaccination with Gudair has recently commenced. This review examines prospects for the control of OJD by chemotherapy, vaccination and mycophages. Current chemotherapeutic regimes for treatment of M. paratuberculosis in ruminants are prohibitively expensive and of dubious efficacy, and apart from environmental concerns, mycophage therapy lacks a track record of success against intracellular bacteria. There is substantial evidence that live and killed mycobacterial vaccines reduce the incidence of clinical disease and shedding rates in OJD. An appraisal of recent experimental results suggests that neonatal vaccination with a defined dose of M. paratuberculosis offers the best prospects for the induction of protective Th1-type immunity.

Animals↗
PubMed2006

Efficacy of a killed vaccine for the control of paratuberculosis in Australian sheep flocks.

A field trial was undertaken from 1999 until 2004 to determine the efficacy of a killed M. a. paratuberculosis vaccine, Gudair, for the control of ovine Johne's disease (OJD) in merino sheep run under Australian pastoral conditions. On each of three farms experiencing significant OJD losses (5-15% per annum), 200 merino lambs (age 1-4 months) were vaccinated, and 200 lambs were left unvaccinated. Animal assessments and sample collections were conducted twice yearly until 4 or 5 years of age. The impact of vaccination on mortality rate, faecal shedding of M. a. paratuberculosis (by pooled and individual faecal culture), liveweight, wool productivity, vaccine injection site lesions and cellular (BOVIGAM) and humoral (PARACHEK) immunity was examined. Vaccination stimulated cell-mediated and humoral immune responses, reduced mortalities due to OJD by 90% and delayed faecal shedding for the first year post-vaccination. Thereafter, the prevalence of shedders among vaccinates was reduced by 90%. The numbers of M. a. paratuberculosis excreted by the vaccinated groups were also reduced by at least 90% at most sampling times. However, high levels of excretion by vaccinates occurred on some occasions, and although only 7 of 600 vaccinates died from OJD, all 7 had multibacillary lesions. Thus there remains a risk that some vaccinated sheep will transfer the disease. Small reductions in liveweight were found in vaccinated lambs in the first year, but there was little effect on wool production. Vaccine injection site lesions were detected in almost 50% of sheep after 2 months, and these persisted for at least 4 years in 20-25% of vaccinates. Data from this trial enabled the registration of Gudair in Australia in 2002 and underpins the pivotal role of vaccination in the current management of OJD.

Animals↗
PubMed2006

Comparative evaluation of two decontamination methods for the isolation of Mycobacterium avium subspecies paratuberculosis from faecal slurry and sewage.

Faecal slurry of animal origin from sale yards and raw sewage from a sewage treatment plant were sampled for the radiometric culture over 5 months at approximately weekly intervals. Before the radiometric culture, samples were decontaminated using the double incubation method. One set of triplicate samples of slurry and sewage was decontaminated at 37 degrees C and the other set was decontaminated at 42 degrees C. M. a. paratuberculosis or its DNA was detected in seven of 45 cultures (15.6%) of slurry decontaminated at 37 degrees C and in 14 of 39 cultures (35.9%) of slurry decontaminated at 42 degrees C. The contamination rates in cultures of slurry processed at 37 degrees C and 42 degrees C were 82.2% and 69.2%, respectively. M. a. paratuberculosis DNA was also detected in one of 45 cultures (2.2%) of sewage decontaminated at 42 degrees C. The contamination rates in samples of sewage processed at 37 degrees C and 42 degrees C were 84.4% and 4.4%, respectively. Results of this study warrant further investigations to evaluate the suitability of a decontamination method at 42 degrees C for the isolation of M. a. paratuberculosis from faeces, tissues and milk.

Animals↗
PubMed2006

Comparison of BACTEC 460 and MGIT 960 systems for the culture of Mycobacterium avium subsp. paratuberculosis S strain and observations on the effect of inclusion of ampicillin in culture media to reduce contamination.

To compare the utility and diagnostic accuracy of BACTEC and MGIT culture systems, a total of 41 pooled faecal samples, each containing faeces from one sheep infected with the S strain of Mycobacterium paratuberculosis and four uninfected sheep was cultured. The MGIT culture system did not support the growth of the S strain of M. paratuberculosis from faeces within the time frame of the experiments, although a laboratory adapted S strain grew slowly in MGIT provided that sufficient bacteria were inoculated. In contrast, C strain grew rapidly in MGIT. The sensitivity of culture was calculated relative to the infection status of the animals, none of which had clinical signs of ovine Johne's disease. The overall sensitivity of pooled faecal culture in the BACTEC culture system was 21.9% (95% confidence limits, 10.5-37.6), a figure dependant on the proportion of multibacillary cases. The sensitivities of the BACTEC culture system for pools containing animals with multibacillary and paucibacillary lesions were 100.0% (95% confidence limits, 47.2-100.0) and 17.8% (95% confidence limits 6.06-36.8), respectively. The contamination rate of BACTEC cultures was 9.7% compared to 14.3% for MGIT. The effect of 100 microg/ml ampicillin on the S strain of the M. paratuberculosis was examined and in both BACTEC and MGIT media it delayed growth by about 1 week. The composition of MGIT medium, particularly presence of vancomycin hydrochloride, slowed the growth of the S strain. The low content of egg yolk was considered to be another possible factor. The radiometric BACTEC culture system remains the best alternative for the culture of S strain and is recommended in circumstances where the genotype (s) of the strains present in a region/farm is either unknown or S strain.

Ampicillin↗
PubMed1996

Description and classification of different types of lesion associated with natural paratuberculosis infection in sheep.

Paratuberculosis lesions in naturally infected sheep were classified histologically. Adult sheep (n = 166) culled for various reasons, from four flocks in which clinical cases of the disease had occurred, were studied. Eight-two sheep (49.4%) showed lesions that could be divided into three main categories. Type 1 lesions, found in 24.1% of these animals, consisted of small granulomata formed by macrophages and were located exclusively in the ileocaecal Peyer's patch. In type 2 lesions, found in 4.8% of the sheep, granulomata were also observed in the mucosa associated with Peyer's patches. Type 3 lesions were characterized by granulomata in areas of the mucosa associated with, and also distinct from, the Peyer's patches. Three subtypes of type 3 lesions were recognized. In subtype 3a, found in 4.2% of the sheep, multifocal granulomata appeared in different areas of the lamina propria; they were not apparently associated with lymphoid tissue, and neither did they modify the morphology of the affected areas. Subtype 3b, found in 13.9% of animals, consisted of large numbers of macrophages, widespread in the lamina propria; in subtype 3c (2.4% of sheep) lymphocytes were the main inflammatory cell, with some macrophages scattered amongst them. In subtypes 3a and b, villi were distended and the mucosa appeared thickened. Mycobacteria could be demonstrated in tissue sections from all the samples with subtype 3b lesions and in almost all of those with type 2 and 3a lesions; these organisms were absent or sparse, however, in type 1 and 3c lesions. Macroscopical lesions were clearly visible only in sheep with type 3b and 3c lesions. Mycobacterium paratuberculosis was cultured from all the animals with type 3 lesions, from 87.5% of those with type 2 lesions, and from 47.5% of those with type 1. Type 3b lesions resembled the "borderline-lepromatous" form of mycobacterial lesions, whereas type 3c lesions resembled the "borderline-tuberculoid" form. The relationship between intestinal lymphoid tissue and paratuberculosis lesions is discussed, and the diagnostic importance of histological examination of the ileocaecal valve emphasized.

Animals↗
PubMed1996

Relationship of acid phosphatase activity to ultrastructural features in mice inoculated with Mycobacterium paratuberculosis.

Macrophage activation, measured as increased acid phosphatase (AcPase)-positive areas by image analysis, and ultrastructural features were examined in granulomatous mycobacterial lesions of mice innately susceptible (BALB/c mice; Bcg) and innately resistant (C3H/HeJ mice; Bcg) to Mycobacterium paratuberculosis strain ATCC 19698. In the liver and spleen of BALB/c mice 3 weeks after intraperitoneal inoculation with M. paratuberculosis, AcPase activity detected in epithelioid cell nodules was high; it had decreased, however, in the liver and spleen after a further 3 and 6 weeks, respectively. In C3H/HeJ mice, the size of epithelioid cell nodules in the liver and spleen was smaller than in BALB/c mice, and infiltrating macrophages, which had increased by week 9 after inoculation, showed high AcPase activity. Ultrastructurally, by week 32 in BALB/c mice, small phagolysosomes (SPLs) had greatly increased in number in the epithelioid cells. These SPLs contained a few AcPase-positive areas and a small number of bacteria, most of which were surrounded by an electron-translucent space (or electron-transparent zone [ETZ]). In contrast, only a few SPLs were observed in C3H/HeJ mice at week 32; in the liver and spleen, large phagolysosomes (LPLs) showed high AcPase activity and contained many degenerated bacteria, which also had an ETZ. These results suggest that the enzymatic and ultrastructural differences in phagolysosomes between BALB/c mice and C3H/HeJ mice reflect the susceptibility of these mouse strains to M. paratuberculosis.

Acid Phosphatase↗
PubMed2002

Localisation of CD25+ cells and MHCII+ cells in lymph nodes draining Mycobacterium avium subsp. paratuberculosis vaccination granuloma and the presence of a systemic immune response.

Vaccination of goat kids against paratuberculosis protects against lesions and clinical disease. The systemic cellular response was studied in goat kids 3-9 weeks after vaccination. Peripheral blood cells showed increased interferon-gamma production and expression of interleukin-2 receptor (CD25) after stimulation with Mycobacterium avium subsp. paratuberculosis antigens. The lymph node draining the vaccination granuloma was studied three weeks after vaccination in a parallel group of goat kids. In deep cortex, MHCII+ cells were observed surrounded by CD4+ T-cells, while follicular hypertrophy and hyperplasia were prominent in the subcapsular region and along connective tissue trabecula. Comparison of the local and systemic immune responses revealed an inverse relationship between CD25+ T-cells in the lymph node deep cortex and cells in peripheral blood that up-regulate CD25 upon in vitro stimulation, suggesting that activated and regulatory T-cells in the local lymph node influence the level of circulating antigen-specific T-cells following vaccination against paratuberculosis in goats.

Animals↗
PubMed2003

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 with respect to transmission of paratuberculosis has yet to be addressed. We selected and bought 16 cows, all proven to be moderate shedders of the bacterium in the faeces immediately prior to the experiment but none were clinically sick. One sample of uterine content was collected from each animal by flushing the uterus on the day of heat and five samples of homogenised uterine tissue were collected on the eighth day of the same cycle by biopsy. In addition, 217 cumulus-oocyte complexes (COCs), ranging from 3 to 35 COCs per animal, were collected using ultrasound guided transvaginal puncture of the ovarian follicles (OPU). On the seventh day of the subsequent cycle 31 embryos were obtained using the classic technique of super ovulation induction, artificial insemination (AI), followed by flushing of the uterus. These embryos have been washed and trypsinised. Fourteen of the 16 cows were treated again for super ovulation in the subsequent cycle and 19 foetuses were collected by opening of the uterus after euthanasia on Days 35-49 of the cycle. All samples were cultured for presence of Map and checked every 2 months during 1 year for bacterial growth. None of the samples showed growth of Map after 12 months of culture. Pathological examination of the cows revealed different degrees of severity of pathological alterations of the intestinal tract and mesenteric lymph nodes. However, the results suggest that neither in vivo embryo's nor oocytes are carriers of the bacteria and do not form an extra risk at transfer. However, due to the limited size of the experiment (sample size of 16 cows), a certain margin for error remains.

Animals↗
PubMed2000

The use of interleukin-2 receptor expression as a marker of cell-mediated immunity in goats experimentally infected with Mycobacterium avium ssp. paratuberculosis.

The purpose of the present work was to demonstrate cell-mediated immune response to paratuberculosis in experimentally infected animals, using quantification of interleukin-2 receptor (IL-2R) expression on activated lymphocytes by means of in vitro stimulation with Mycobacterium avium ssp. paratuberculosis-derived purified protein derivative (PPDp). A whole-blood technique was developed, and optimal conditions for quantification of IL-2R expression on caprine lymphocytes, using monoclonal antibodies (anti-bovine IL-2R-alpha) and low cytometrical analysis, were determined. Different PPDp-antigen concentrations and incubation times were compared. The whole-blood method was also compared to the more traditional IL-2R assay using peripheral blood mononuclear cultures (Hesketh et al., 1993). Cross-reactivity to Mycobacterium avium was studied at different mycobacteria-PPD concentrations. An immune response could be demonstrated in animals infected with Mycobacterium avium ssp. paratuberculosis. We found that a PPDp concentration of 10microgml(-1) together with an incubation time of 72h, gave the best results using the whole-blood method. The whole-blood method eliminates many laborious steps involved in lymphocyte separation, and the effects of all the constituents of blood are expressed in a way which corresponds more to in vivo conditions. The risk of selecting subpopulations of lymphocytes during cell separation is avoided.

Animals↗
PubMed2002

In vitro responses to purified protein derivate of caprine T lymphocytes following vaccination with live strains of Mycobacterium avium subsp paratuberculosis.

Live attenuated vaccines provide protection against intestinal lesions in goats infected with Mycobacterium avium subsp. paratuberculosis. To examine the role of different T lymphocyte subsets in the development of this protective immunity, CD4(+), CD8(+) and gamma delta T cell receptor (TCR)(+) cells from peripheral blood of goat kids vaccinated with live attenuated strains of M. a. paratuberculosis were studied. After in vitro stimulation with purified protein derivate, the expression of gamma-interferon (IFN-gamma) and the activation marker interleukin-2 receptor (IL-2R) was analysed by flow cytometry. A depletion experiment was performed, where the phenotypes and IL-2R expression was studied after stimulation of cultures depleted of a T lymphocyte subpopulation. Close to all of the IFN-gamma producing cells were of the CD4(+) subset, while only a small number were CD8(+) cells. The gamma delta TCR(+) cells were highly activated, but did not produce IFN-gamma after in vitro stimulation. Depletion of CD4(+) cells lead to a decrease in the percentage of total gamma delta TCR(+) cells and gamma delta TCR(+)IL2-R(+) cells. Removing the gamma delta TCR(+) cells increased the relative numbers of CD4(+), but not the CD4(+)IL-2R(+) cells. Insight into the in vitro recall responses of T cell subsets from animals vaccinated with live paratuberculosis vaccines is essential in the development of more efficient vaccines.

Animals↗
PubMed2000

Prevalence of paratuberculosis (Johne's disease) in the Belgian cattle population.

The national bovine paratuberculosis (PTB) seroprevalence (apparent prevalence) in the Belgian cattle population was determined by a serological survey that was conducted from December 1997 to March 1998. In a random sample of herds (N=556, 9.5%), all adult cattle of 24 months of age or older (N=13,317, 0.4%) were tested for the presence of antibodies using a commercially available absorbed ELISA test kit. The PTB median within-herd seroprevalence (proportion of detected animals within the seropositive herds) and the PTB individual-animal seroprevalence (proportion of detected animals) were, respectively, 2.9% (quartiles=1.6-5.6) and 0.87% (95% confidence interval (CI)=0.71-1.03). The PTB herd seroprevalence (proportion of detected herds) was 18% (95% CI=14-21). Assuming a test sensitivity and specificity of 45 and 99% [Sweeney et al., 1995. J. Vet. Diagn. Invest. 7 (4), 488; Sockett et al., 1992. J. Clin. Microbiol. 30 (5), 1134], respectively, the median true within-herd prevalence and the true individual-animal were estimated to be 7 and 2%, respectively. The true herd prevalence of Mycobacterium paratuberculosis infection was first estimated according to currently accepted methodology. This calculation revealed that the specificity of the used test has a dramatic effect on the estimation; assuming a test sensitivity of 45% and a true within-herd prevalence of 7%, the true herd prevalence estimation decreased from 36 to 0.8% if the test specificity decreased from 99. 9 to 99%, respectively. This sensitivity analysis showed that the practical limits of the accuracy of the used screening test jeopardize the estimation of the true herd prevalence within reasonable confidence limits, because the within-herd PTB true prevalence was low. For this reason we augmented the herd specificity for herds with larger adult herd size (>5). This was done by increasing the cut-off number of positive cattle required (>/=2) to classify a herd truly positive and including herds with one positive test result if there was historical evidence of PTB (previous diagnosis and/or clinical signs). This approach resulted in an estimated true herd prevalence of M. paratuberculosis infection of 6%. The true herd prevalence for dairy, mixed and beef herds was, respectively, 10, 11 and 3%.

Animals↗
PubMed2000

Modification of a bovine ELISA to detect camelid antibodies to Mycobacterium paratuberculosis.

Mycobacterium avium subsp. paratuberculosis infection, or Johne's disease, reportedly has a low prevalence in South American camelid populations. Recently, however, single cases in the United States as well as an outbreak of the disease in Australian alpacas (Lama pacos) have been described. To provide a rapid and cost-effective method of diagnosing Johne's disease in this species, the bovine Parachek((R)) Johne's Absorbed EIA (CSL, Vic., Australia) was modified to create a camelid-specific serum antibody assay. An anti-llama IgG conjugated to horseradish peroxidase replaced the anti-bovine immunoglobulin. Checkerboard titration of principal reagents was performed using serum from nine tissue and/or fecal culture-positive camelids. Optimal dilutions of key components were determined in order to provide clear discrimination between positive and negative controls. Completion of a kinetic assay determined the optical density at which the enzyme-substrate reaction should be stopped. A herd of 100 camelids with no history of disease or exposure to M. a. paratuberculosis, a subset of which were tissue and/or fecal culture-negative, was tested to establish a cut-off value. Sample results were expressed as a percentage of the results for control sera by transforming optical density values to ELISA values (EV%). A preliminary EV% cut-off of 20 was established. Using this prototype assay, culture-positive animals showed significantly different antibody responses from culture-negative animals. These results indicate that this camelid-specific ELISA, once refined, may be a useful tool for screening camelid herds for M. a. paratuberculosis infection.

Animals↗
PubMed2000

Mycobacterium avium subsp. paratuberculosis infection in cattle in Austria, diagnosis with culture, PCR and ELISA.

Serum samples from healthy, infected (n=11) and diseased (n=2) cattle as well as positive (n=17) and negative (n=41) reference sera were tested for antibodies to Mycobacterium avium subsp. paratuberculosis with two ELISA-methods (A-ELISA, Allied Monitors, Fayette, USA; H-ELISA, Institute of Microbiology and Animal Diseases, Veterinary University Hannover). Fecal samples of these animals were examined by PCR and culture. Also field serum samples found to be positive (n=664) or inconclusive (n=1589) by A-ELISA during a survey done on 11028 cattle of 2757 farms at different districts in Austria were retested with H-ELISA (Gasteiner et al., 1999). In both ELISA-methods total agreement between antibody detection and shedding of M. avium subsp. paratuberculosis (PCR, culture) in cases of diseased animals during the testing period was found. In subclinically infected animals H-ELISA showed a better correlation with the results of PCR and fecal culture. Reference serum samples of culturally negative cattle were negative in 98% by H-ELISA and in 82% by A-ELISA, and those of positive animals were positive in 59% by H-ELISA and in 82% by A-ELISA. The 664 A-ELISA positive field serum samples were positive in 20.5%, inconclusive in 32.5% and negative in 47% by H-ELISA. A-ELISA inconclusive sera gave positive reactions by H-ELISA in 5.2%, negative in 74.8% and inconclusive results in 20%. The highest prevalence of antibodies (7.9% by A- and 2.2% by H-ELISA) against M. avium subsp. paratuberculosis were found in cattle at the age of six and seven years. Seropositive animals were found at all tested ages. The A-ELISA gave two to three times more positive reactors than the H-ELISA. Also both tests showed the highest prevalence of reagents among Holstein Friesian (6.2% by A-ELISA, 2.5% by H-ELISA) followed by other cattle breeds. Seropositive cattle were observed in all districts of Austria in 3. 3-7.1% and in 0.5-1.8% of herds according to A- and H-ELISA, respectively.

Animals↗
PubMed2000

Use of a PCR method on fecal samples for diagnosis of sheep paratuberculosis.

The high sensitivity of PCR compared to the difficulties of fecal culture in sheep prompted the development of PCR protocols for detection of Mycobacterium avium subsp. paratuberculosis DNA in sheep feces. Although the PCR itself is well developed, and does not pose large technical problems, concentrating the bacteria from samples that may contain low numbers of bacilli using practical methods is still the main difficulty for the use of this technique. In this study, we describe an extraction protocol for the concentration and purification of M. avium subsp. paratuberculosis DNA from fecal samples and we compare it with other methods. The diagnostic performance of the freeze-boiling method was evaluated using a reference method [Vet. Rec. 134 (1994) 95] on fecal samples from a group of selected sheep from different flocks of known individual serological, pathological, and cultural paratuberculosis status. Using, as a reference, a combination of results in those conventional methods, the freeze-boiling PCR protocol showed a sensitivity of 94.1%, and a specificity of 92.3%.

Animals↗
PubMed2000

Transitions in immune responses to Mycobacterium paratuberculosis.

The host immune response to infection with Mycobacterium paratuberculosis is paradoxical, with strong cell-mediated immune responses during the early, subclinical stages of infection and strong humoral responses during the late clinical stages of the disease. Cell-mediated immune responses modulated by various T cell subsets are essential to provide protective immunity and prevent progression of the disease. Secretion of cytokines by T cell populations serves to activate macrophages to kill ingested M. paratuberculosis as well as activate other T cell subsets to contain the infection. This paper reviews the current knowledge of T cell immune responses in M. paratuberculosis infection based upon clinical studies and research using mouse models.

Animals↗
PubMed2002

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 serological tests: a complement fixation test (CFT), agar gel diffusion test (AGID) and enzyme-linked immunosorbent assay (ELISA), as well as an IFN-gamma test on samples of blood. Among the 19 inoculated sheep, 16 gave positive reactions in the PLN IFN-gamma assay on samples incubated overnight, and 18 tested positive when the assay was applied to PLN samples incubated for 48h. In comparison, three, four and seven inoculated sheep gave positive reactions in the ELISA, CFT and in the blood IFN-gamma assay on samples incubated overnight, respectively. The AGID and IFN-gamma assay on blood samples incubated for 48h detected eight inoculated animals. Twelve inoculated sheep, that tested positive in the PLN IFN-gamma assay were clinically normal, gave negative results in an IS900-based polymerase chain reaction (PCR) assay on samples of ileum and ileocaecal lymph node and had no histological evidence of paratuberculosis, but tested positive on more than two occasions in sequential serological testing before necropsy. None of the 10 non-inoculated sheep tested positive in the AGID, CFT, ELISA, blood IFN-gamma assay on samples incubated overnight and for 48h or the PLN IFN-gamma assay on samples incubated overnight, but one gave a positive result in the PLN IFN-gamma assay on samples stimulated for 48h. It is likely that the positive reactions obtained by the PLN IFN-gamma assay in the 12 inoculated sheep that tested negative in the PCR assay and histopathological examination represents immunological evidence of latent infection or previous exposure to M. paratuberculosis rather than active infection.

Animals↗
PubMed1997

Development of an ELISA technique for serodiagnosis of bovine paratuberculosis.

A local clinical Mycobacterium avium ssp. paratuberculosis (M. paratuberculosis) isolate was cultivated in large-scale culture. A procedure for efficient preparation of a lipoarabinomannan-containing antigen was developed and standardized; 25 mg of purified antigen were obtained per gram of bacterial wet weight. An ELISA based on this antigen was developed. Intra- and interassay variation were determined to be 20% and 27%, respectively. The ELISA was evaluated using the sera of groups of 39 non-randomly selected and 92 randomly selected animals from which ileocaecal lymph nodes were cultured to isolate viable M. paratuberculosis. Combining the results of both groups a positive predictive value of 74% and a negative predictive value of 99% were calculated. The ELISA was licensed by the German regulatory agencies and, in a direct comparison, was found to be superior to both other licensed assays.

Animals↗
PubMed1999

Pathogenesis of Mycobacterium avium subspecies paratuberculosis infections in ruminants: still more questions than answers.

Mycobacterium avium subspecies paratuberculosis is the etiologic agent of paratuberculosis (Johnes disease), a chronic enteritis in ruminants, which is one of the most widespread bacterial diseases of domestic animals, causing enormous economic losses worldwide. Though the disease was first described more than a century ago, the biology of the infecting organism and the mechanisms of its interactions with the host still remain a mystery. In this review, recent advances made on pathogenesis of paratuberculosis are summarized and future challenges are discussed.

Animals↗