Mycobacterium Paratuberculosis Infection Research Articles

Investigation of Mycobacterium paratuberculosis in Arabian dromedary camels (Camelus dromedarius).

Aim:Mycobacterium avium subspecies paratuberculosis (MAP) causes Johne’s disease in ruminants. This study aimed to investigate Mycobacterium paratuberculosis infection in clinically infected camels on the immunological, conventional bacteriological, and molecular biological basis.Materials and Methods:A total of 30 Arabian camels (Camelus dromedarius) were examined in this study. The camels were suffering from signs ranging from mild to severe infections (that did not respond to antibiotic treatment) to chronic or intermittent diarrhea. Camels were grouped into three groups based on their age, sex, and breed. Detection of anti-MAP antibodies in camels’ serum, Ziehl–Neelsen (ZN) staining technique on rectal scraps, direct recognition of MAP in stool and tissue specimens by IS900 polymerase chain reaction (PCR) assay, and finally isolation and molecular description of MAP from fecal and tissue samples were carried out.Results:Five MAP isolates were recovered from these investigated camel samples giving an isolation rate of 16.6%, while eight camels were identified by PCR (26.6%). Five camels yielded MAP in their feces by ZN fecal staining (16.6%), whereas ELISA detected anti-MAP antibodies in nine camels only (30%).Conclusion:From the obtained results, we concluded that the gold standard for the diagnosis of MAP is the culture method despite its limitations. Molecular diagnosis (PCR) could be a useful tool in the identification of truly positive and negative camels; however, great care should be given regarding the primers specificity and sensitivity.

Possible association between Mycobacterium paratuberculosis infection and Crohn's disease in human

Crohn's disease is a granulomatous ileocolitis of humans, of unknown aetiology, which generally manifests itself during the prime of life. The chronic, progressive clinical course and histological findings are consistent wiht a mycobacterial aetiology. Evidence supporting a pathogenic role for a mycobacterium has become available only in the last decade with the isolation of this microorganism from Crohn's disease tissue. M. paratuberculosis, which is the causative agent of Johne's disease in animals, has been identified in patients with Crohn's disease by PCR and DNA hybridisation techniques. It has been shown that isolates of M. paratuberculosis from Crohn's disease are indentical with pathogenic strains in ruminants.

Loss of income from cows shedding Mycobacterium avium subspecies paratuberculosis prior to calving compared with cows not shedding the organism on two Minnesota dairy farms

Quantification of the financial effect of Mycobacterium paratuberculosis infection on lactation performance is essential to encourage participation of dairy cattle producers in Johne's disease (JD) control programs. The objective of this study was to evaluate the differences in net income per lactation of cows shedding Mycobacterium paratuberculosis before calving compared with test-negative cows. Two Minnesota dairies were enrolled in the study and fecal samples were collected from 1,048 cows during the close-up period. Milk production, clinical diseases (other than clinical JD), and reproductive performance data were recorded for each cow. Overall, fecal-culture-positive (FCP) cows produced 1,355kg less than fecal-culture-negative (FCN) cows. Fecal-culture-positive cows that survived their current lactation produced $276 less in milk income than cows that were FCN ($1,956 vs. $1,680; SD $526, $570). Fecal-culture-positive cows were 3.0 (95% confidence interval: 1.6-5.8) times more likely to be culled than FCN cows. The mean days open (number of days from calving to conception) was not statistically significant and the cost differences for clinical disease other than JD were small and neither statistically nor economically significant between FCP and FCN cows. Among all FCP cows, income over feed costs losses were $366 per cow per lactation compared with FCN cows. Among FCP nonculled cows, income over feed costs losses were $276 more compared with FCN cows and this difference was statistically significant. There was a total loss of $155 per lactation for nonculled FCP cows retained in the herd compared with FCN cows retained in the herd. Among culled cows, FCP cow losses were $50 less because of age at culling and $120 for reduced beef value. This totaled a loss of $441 for culled FCP cows compared with culled FCN cows. The losses as a result of lower lactation performance and early culling from the herd should alarm dairy producers and motivate them to implement the appropriate control measures for the disease.

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 control subjects, using three antigens attributed with species-specificity and selective immunogenicity following MAP infection. Antibodies binding to the 38-kD band of MAP extract were demonstrable by the Western blot technique in 57% of CD patients. Antibody levels to the 24-kD (p24BCD) cathodic bands, determined by competition ELISA using a monospecific murine antiserum, and to the 18-kD protease-resistant purified bacterioferritin, detected by standard ELISA, were significantly elevated in 53% of CD patients. However, these three antibody specificities tested in individual CD patients did not show any correlation with each other. Thus, 18% of patients were positive for all three specificities, whilst 84% had antibodies to at least one of the specific antigens. Although the exact proportion of affected patients is yet to be defined, the serological results obtained support the view that MAP infection may play an etiological role in Crohn's disease.

Effects of Shipping and Storage Conditions of Fecal Samples on Viability of Mycobacterium paratuberculosis

Johne's disease is characterized by chronic granulomatous enteritis of ruminants caused by Mycobacterium paratuberculosis infection. Early diagnosis of infected animals is necessary to reduce contamination of the environment and to control disease spread through feces ([3][1], [6][2]). About 75% of

Identification of a QTL on BTA20 affecting susceptibility toMycobacterium aviumssp.paratuberculosisinfection in US Holsteins

The objective of this study was to identify QTL affecting susceptibility to Mycobacterium paratuberculosis infection in US Holsteins. Twelve paternal half-sib families were selected for the study based on large numbers of daughters in production and limited relationships among sires. Serum and faecal samples from 4350 daughters of these 12 sires were obtained for disease testing. Case definition for an infected cow was an ELISA sample-to-positive ratio >/=0.25, a positive faecal culture or both. Three families were selected for genotyping based on a high apparent prevalence (6.8-10.4% infected cows), high faecal culture prevalence (46.2-52.9% positive faecal cultures) and large numbers of daughters tested for disease (264-585). DNA pooling was used to genotype cows, with an average of 159 microsatellites within each sire family. Infected cows (the positive pool) were matched with two of their non-infected herdmates in the same lactation (the negative pool) to control for herd and age effects. Eight chromosomal regions putatively linked with susceptibility to M. paratuberculosis infection were identified using a Z-test (P < 0.01). Significant results were more rigorously tested by individually genotyping cows with three to five informative microsatellites within 15 cM of the significant markers identified with the DNA pools. Probability of infection based on both diagnostic tests was estimated for each individual and used as the dependent variable for interval mapping. Based on this analysis, evidence for the presence of a QTL segregating within families on BTA20 was found (chromosome-wide P-value = 0.0319).

Effect of Mycobacterium paratuberculosis infection on production, reproduction, and health traits in US Holsteins

Our objective was to estimate the effect of Mycobacterium paratuberculosis infection on milk, fat, and protein yield deviations, pregnancy rate, lactation somatic cell score, and projected total months in milk (productive life). A serum ELISA and fecal culture for M. paratuberculosis were performed on 4375 Holsteins in 232 DHIA herds throughout the US. Primarily first through third lactation cows (99% of total) were assayed for infection. Trait information (except productive life) was obtained for the lactation concurrent with disease tests. Productive life was total months in milk through a cow's life, which was projected if a cow was still milking. For most analyses, case definition for M. paratuberculosis infection was defined as either an ELISA S/P ratio ≥ 0.25 or a positive fecal culture for M. paratuberculosis or both. To determine if diagnostic test affected estimates, case definition was redefined to include only cows with ELISA S/P ratios ≥ 0.25 or only fecal culture-positive cows. Linear models were used to estimate effect of M. paratuberculosis infection on traits. M. paratuberculosis-infected cows (7.89% of cows) produced 303.9 kg less milk/lactation, 11.46 kg less fat/lactation, and 9.49 kg less protein/lactation ( P ≤ 0.003). Infected cows had higher pregnancy rates (1.39%) ( P = 0.0385) and lower productive life (2.85 months) ( P ≤ 0.0001). M. paratuberculosis infection did not affect somatic cell score. Effect of infection on milk and protein yields was larger in first lactation M. paratuberculosis-positive cows relative to cows that tested positive in later lactations. Fecal culture-positive cows had consistently larger effects on all traits than ELISA-positive cows. M. paratuberculosis infection, and not just clinical Johne's disease, decreases milk, fat, and protein yields, thus increasing the estimated cost of paratuberculosis to the US dairy industry.

THE PREVALENCE OF MYCOBACTERIUM PARATUBERCULOSIS INFECTION IN ILEOCECAL VALVE OF CATTLE SLAUGHTERED IN AHVAZ ABATTOIR, SOUTHERN

*Summary To investigate the prevalence of Mycobacterium paratuberculosis (M. avium subsp. paratuberculosis) infection among cattle slaughtered in Ahvaz abattoir, samples were taken from 250 cattle. Before slaughter, sex and age were assessed. Thereafter, samples were taken from ileocecal valve, rectum mucosa, ileocecal lymph node and stool. All smears which taken from these samples were stained by Ziehl-Neelsen method and examined by light microscopy. At first, ileocecal samples were examined; other samples were examined when ileocecal sample was positive. Out of the 250 cattle, 5 (2%) were positive. With the exception of one sample in which the organism was observed in both ileocecal valve and ileocecal lymph node, in other positive samples the bacteria was observed in only ileocecal valve smear.

Evaluation of a Pourquier ELISA kit in relation to agar gel immunodiffusion (AGID) test for assessment of the humoral immune response in sheep and goats with and without Mycobacterium paratuberculosis infection

The present study was designed to evaluate a commercial ELISA kit (Institut Pourquier) for the diagnosis of ovine and caprine paratuberculosis under Australian conditions and to compare its accuracy with the existing AGID test. The sensitivity of the ELISA in sheep and goats was 34.9% and 56.4%, with a specificity of 98.8% and 100.0%, respectively. Sensitivity of AGID was 13.8% for sheep and 39.5% for goats, with specificity of 100.0% for both species. The sensitivity of the ELISA in sheep depended on the category of histological lesions. AGID and ELISA were conditionally independent, and appeared to detect overlapping but distinct subgroups of infected animals. The ELISA was significantly more sensitive than the AGID. The ELISA was simple to perform, robust and repeatable. Coefficients of variation of <12.0% were observed for positive and negative controls included on 193 plates over a 10-month period and there was a high level of intraassay repeatability with 12.0% of the duplicate samples having CV of >15.0%.

Certification of herds as free of Mycobacterium paratuberculosis infection: actual pooled faecal results versus certification model predictions

Dutch dairy herds closed for at least 3 years with no history of paratuberculosis were recruited for a study on herd-certification. One hundred dairy herds were tested for Mycobacterium paratuberculosis at 6-month intervals by pooled faecal culture (five individual animal samples per pool) with solid media. Ninety of the herds completed 9 herd tests and 10 herds dropped out of the study for reasons other than a paratuberculosis diagnosis. Of the 90 herds completing the full study, 61% eventually were found to be M. paratuberculosis-infected. The number of infected herds detected decreased with each round of testing. Assuming that all infected herds had been detected by the ninth herd test, the observed percentage of herds that were truly noninfected (P-free) after each round of testing was calculated. The observed P-free was compared to the predicted P-free based on a previously reported herd-certification model. The P-free predicted by the model was significantly different from the observed P-free. When a single assumption in the model was changed and a diagnostic sensitivity of 40–50% was selected, the predicted P-free closely approximated the observed P-free for the 90 Dutch dairy herds studied. The critical assumption that was changed for Version 2.0 of the model was within-herd infection prevalence for infected but test-negative herds after each round of serial testing. Model Version 1.0 had assumed a 50% decrease in within-herd prevalence but Version 2.0 assumed a stable within-herd prevalence. Culture of pooled faecal samples provides a high-sensitive, high-specific, low-cost test for herd-certification programs.

The Effect of Subclinical Mycobacterium paratuberculosis Infection on Milk Production in Michigan Dairy Cows

The objective of this study was to determine the effect of subclinical Mycobacterium paratuberculosis infection on mature equivalent milk, protein, and fat production in a sample of Michigan dairy herds with a history of cows positive for M. paratuberculosis diagnosed by fecal culture. A prospective two-group cohort study was conducted. Participating herds were tested, and productivity and reproduction records were monitored for 18 mo. All cows aged 24 mo and greater were tested for M. paratuberculosis infection using the ELISA and radiometric fecal culture (RFC) techniques. Using both tests in parallel, the overall sample apparent prevalence for M. paratuberculosis infection was 41.8%. Adjusting for diagnostic sensitivity and specificity resulted in a calculated sample true prevalence of 59.9%. Subclinical paratuberculosis test-positive status had no statistically significant effect on mature equivalent milk, fat, or protein production. The results of this study concur with the findings of other studies, reporting that the magnitude and direction of the association between subclinical paratuberculosis infection and milk production depends upon the parity of the animal, stage of disease, and the stage in lactation being monitored. Assessment of the impact of subclinical paratuberculosis on milk production must consider the average parity of the sample population. In herds that have an average parity of 2 or less, subclinical paratuberculosis infection may have little impact on milk production.

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%.

The effect of subclinical Mycobacterium paratuberculosis infection on days open in Michigan, USA, dairy cows

A prospective cohort study design was used to evaluate the impact of subclinical Mycobacterium paratuberculosis infection on days open in a sample of Michigan dairy herds with a history of cows positive for M. paratuberculosis diagnosed by fecal culture. Participating herds were tested and productivity and reproduction records were monitored for 18 months. All cows ≥24 months old were tested for M. paratuberculosis infection using the ELISA and radiometric fecal-culture (RFC) techniques. Test-negative cows were re-tested at the conclusion of the monitoring period. Multivariable regression models were used. Using both tests in parallel, the overall sample apparent prevalence for M. paratuberculosis infection was 41.8% (223/533 animals tested). Adjusting for diagnostic sensitivity and specificity, this resulted in a calculated sample true prevalence of 59.9%. ELISA-positive cows (on average) had a 28-day increase in days open when compared to ELISA-negative cows ( p=0.02). The diagnostic method used to define a case altered the apparent association between paratuberculosis test status and days open. Fecal culture was a less-effective diagnostic tool for use in herds with a high prevalence of infected animals. The increase in days open in the ELISA-positive cows was an indication that perhaps reduced estrus expression or an increased post-partum anestrous period occurred in the subclinically infected ELISA-positive animals. This might have been due to a negative energy balance associated with M. paratuberculosis infection.

Dietary calcium modulates Mycobacterium paratuberculosis infection in beige mice

A 6-month study was conducted to evaluate the effects of feeding different levels of dietary calcium (Ca) on the persistence of Mycobacterium paratuberculosis infection using a mouse model. Beige mice, averaging 8 weeks of age, were randomly assigned to one of the following dietary treatments: 1) 0.02% Ca, 2) 0.15% Ca, 3) 0.45% Ca, and 4) 1.0% Ca. Mice were infected intraperitoneally with 10 8 CFU viable M. paratuberculosis for 1, 3, and 6 month periods. Plasma Ca levels was unaffected by dietary Ca ( x = 7.3 mg/dl). Plasma levels of 1,25(OH) 2D 3 was elevated significantly in 0.02% and 0.15% Ca groups compared to other treatments at the end of each period, with the highest levels observed for 0.02% Ca mice and intermediate values for 0.15% Ca mice. One month after infection, numbers of viable M. paratuberculosis cultured from the spleen were significantly reduced for 0.15% Ca mice, whereas the number of bacteria isolated from the liver and mesenteric lymph node (MLN) were higher for the 0.02% Ca group. There were no differences in bacterial numbers in the ileum although they tended to be higher for the 0.02% Ca group. Three months after infection, bacterial numbers in the spleen, ileum, and MLN did not differ across treatments, however, significantly lower numbers were found in the liver of 1.0% Ca mice. Reduced bacterial counts were also observed in the liver of 0.15%, 0.45%, and 1.0% Ca mice after a 6-month infection period compared to the 0.02% Ca group, with the lowest numbers isolated from the 1.0% Ca mice. Numbers of viable bacteria cultured from the ileum and MLN after 6 months of infection were also significantly reduced in 1.0% Ca mice. These results suggest that Ca metabolism is an important modulator of M. paratuberculosis infection.

Farm factors associated with the presence of Mycobacterium paratuberculosis infection in dairy herds on the New York State Paratuberculosis Control Program

An extensive questionnaire was developed and used to collect data from 33 herds that were on the New York State Paratuberculosis Control Program, to study farm factors associated with the presence of Mycobacterium paratuberculosis infection in dairy herds. The results of the last whole herd paratuberculosis fecal culture were used to indicate presence of infection in a herd, with herds having one or more animals positive classified as ‘infected’. The average prevalence within herds was 5.2%. Fourteen herds were uninfected and 19 herds had prevalences ranging from 0.7%–28.2%. Data on 31 continuous and 67 categorical risk factors were collected by questionnaire. Ten factors were significantly associated with prevalence risk of infection in the univariable logistic regression. These factors were: the type of farm operation (commercial/registered or both); earlier diagnosis of the disease before entering the control program; number of clinical cases in the previous year; whether clinical cases were raised or purchased animals; typical signs in clinical cases; exposure of calves 0–6 weeks of age to feces of adult cows; contact of young stock with adult animal feces from using the same equipment to clean the housing for both groups of animals; spreading feces on fields from which forage is later harvested and fed to animals of any age group; what is done with animals that are suspected of having paratuberculosis or test positive on culture; and frequency of cleaning the cow barn. Stepwise logistic regression was used to determine the significance of each risk factor while controlling simultaneously for the effect of other factors. The significant factors were the type of farm operation, clinical signs, and exposure of calves to feces of adult cows. Commercial herds, presence of clinical signs typical of paratuberculosis in animals, and exposure of calves 0–6 weeks old to feces of adult cows all indicate a higher likelihood that a herd is infected with M. paratuberculosis.

Influence of vitamin D3 infusion and dietary calcium on secretion of interleukin 1, interleukin 6, and tumor necrosis factor in mice infected with Mycobacterium paratuberculosis

Abstract Objective To study the effects of 1,25(OH)2D3 and calcium (Ca) on splenocyte cytokine secretion during Mycobacterium paratuberculosis infection. Design Mice were assigned to the following treatments: 1—noninfected, 2—infected, 3—noninfected/1,25-(OH)2D3, 4—infected/1,25(OH)2D3, and 5—infected/low-Ca diet (0.15%). Animals Male beige mice averaging 6 weeks of age and 20 g in body weight. Procedure After acclimation to their diets, mice in treatments 2, 4, and 5 were inoculated IV with 108 colony-forming units of M paratuberculosis. At 1, 6, and 12 months after infection, mice in treatment groups 3 and 4 had miniosmotic pumps implanted subcutaneously that delivered 13 ng of 1,25(OH)2D3/day for 14 days. Treatment 5 was included as a control for comparison with treatment 4 because low dietary Ca should increase endogenous 1,25(OH)2D3 values. Splenocytes were isolated from mice at 1, 6, and 12 months and stimulated in vitro with medium alone (nonstimulated), lipopolysaccharide (LPS), concanavalin A, and M paratuberculosis whole-cell sonicate (MpS). Results Production of interleukin 6 after stimulation with LPS, concanavalin A, or MpS was higher (P &lt; 0.05) for splenocytes isolated from mice fed the low-Ca diet, compared with control infected mice 1 and 6 months after infection. Interleukin 1 and tumor necrosis factor activities were increased (P &lt; 0.05) in splenocytes cultured with LPS and MpS after isolation from mice of the low-Ca group. Mice infused with 1,25(OH)2D3 had higher (P &lt; 0.05) interleukin 1 secretion after stimulation of splenocytes with LPS and higher (P&lt; 0.05) tumor necrosis factor production after incubation with MpS. Conclusion 1,25(OH)2D3 and low dietary Ca increase cytokine secretion in mice infected with M paratuberculosis. (Am J Vet Res 1996;57:825–829)

Associations between subclinical paratuberculosis and milk production, milk components, and somatic cell counts in dairy herds

Objective To determine associations between subclinical Mycobacterium paratuberculosis infection and milk production, milk components, and somatic cell counts of dairy cattle. Design Cross-sectional epidemiologic survey. Animals 23 dairy herds in Wisconsin containing 1,653 adult cows were studied. The herds had above average milk production and a history of bovine paratuberculosis in the herd within the previous 12 months. Procedure All adult cows in the herds were tested for paratuberculosis by use of an absorbed ELISA. Milk yield, fat, protein, and somatic cell count data were retrieved electronically from Dairy Herd Improvement Association records. Results 147 ELISA-positive and 1,506 ELISA-negative cows were identified. ELISA-positive cows had a mature-equivalent milk production of 376 kg (829 lb)/lactation less than that for ELISA-negative herdmates. Significant difference was not found in lactation average percentages of fat and protein, or somatic cell count linear score. When comparing ELISA-positive and -negative cow's current mature equivalent milk with all previous lactations, significant difference was found only from the immediate-preceding lactation. When this difference was examined by parity group, significant difference was confined to cows in the second lactation. Clinical Implications Subclinical paratuberculosis infections, as determined by ELISA, are associated with a 4% reduction in milk yield and add to the already substantial costs of clinical M paratuberculosis infection in the dairy industry. (J Am Vet Med Assoc 1996;208:1872-1876)

Low calcium diet and 1,25-dihydroxyvitamin D 3 infusion modulate immune responses during Mycobacterium paratuberculosis infection in geige mice

A 12-month study was conducted to evaluate the effects of feeding a low calcium (Ca) diet or 1,25-dihydroxyvitamin D 3 (1,25(OH) 2D 3) infusion on the persistence of Mycobacterium paratuberculosis infection using a mouse model. Male beige mice 6–8 weeks of age were assigned to one of the following treatments: (1) non-infected, (2) infected, (3) non-infected/1,25(OH) 2D 3, (4) infected/1,25(OH) 2D 3, and (5) infected/low Ca (0.15%) diet. Infected mice were inoculated intravenously with live M. paratuberculosis. At 1, 6 and 12 months postinfection, mice in Treatments 3 and 4 were implanted subcutaneously with mini-osmotic pumps to deliver 1,25(OH) 2D 3. Infusion with 1,25(OH) 2D 3 exacerbated M. paratuberculosis infection in most tissues at all time points. Mice infused with 1,25(OH) 2D 3 had higher bacterial counts in spleen, liver, and ileum compared with control infected mice after 1 month of infection. In contrast, feeding a low Ca diet reduced the number of viable organisms cultured from the liver and ileum of infected mice. Plasma Ca and 1,25(OH) 2D 3 were increased in mice infused with 1,25(OH) 2D 3 at all time points but values for low Ca mice were not different than for non-infused mice. Splenocyte production of TNF, IL-1 and IL-6 was higher for mice fed the low Ca diet compared with control infected mice after 1 month of infection. Inducible IL-6 activity remained higher for this treatment at 6 months postinfection. These results suggest that feeding a low Ca diet to mice chronically infected with M. paratuberculosis appears to enhance their ability to clear the infection in a manner distinct from any effect of 1,25(OH) 2D 3.

Isolation of Mycobacterium paratuberculosis from colostrum and milk of subclinically infected cows

SUMMARY Mycobacterial culture was performed on colostrum, milk, and feces from 126 clinically normal cows of a single herd with high prevalence of Mycobacterium paratuberculosis infection. Thirty-six (28.6%) cows were determined to be shedding the organism in the feces. Of the 36 fecal culture-positive cows, M paratuberculosis was isolated from the colostrum of 8 (22.2%) and from the milk of 3 (8.3%). Cows that were heavy fecal shedders were more likely to shed the organism in the colostrum than were light fecal shedders.

Prolonged remission in Crohn's disease following therapy for mycobacterium paratuberculosis infection : [formula omitted], M.T. Al-Assi, M. Robinson. Depts. of Medicine, VAMC and Baylor College of Medicine, Houston, TX and University of Oklahoma College of Medicine, Oklahoma City, OK

Prolonged remission in Crohn's disease following therapy for mycobacterium paratuberculosis infection : [formula omitted], M.T. Al-Assi, M. Robinson. Depts. of Medicine, VAMC and Baylor College of Medicine, Houston, TX and University of Oklahoma College of Medicine, Oklahoma City, OK