Environmental contamination with Mycobacterium avium subsp. paratuberculosis in endemically infected dairy herds

Environmental contamination with Mycobacterium avium subsp. paratuberculosis (MAP) is thought to be one of the primary sources of infection for dairy cattle. The exact link between fecal shedding of MAP by individual cows and environmental contamination levels at the herd level was explored with a cross-sectional analysis of longitudinally collected samples on 3 dairy farms. Composite samples from multiple environmental sites in 3 commercial dairy herds in the Northeast US were cultured quarterly for MAP, providing 1131 samples (133 (11.8%) were culture-positive), and all adult animals in the herds were tested biannually by fecal culture (FC), for 6 years. Of the environmental sites sampled, manure storage areas and shared alleyways were most likely to be culture-positive. Environmental sample results were compared to FC results from either the concurrent or previous sampling date at both the herd and the pen level. At the herd level, a 1 log unit increase in average fecal shedding increased the odds of a positive non-pen environmental sample by a factor of 6 and increased the average amount of MAP in non-pen samples by 2.9 cfu/g. At the pen level, a 1 log unit increase in average fecal shedding in the pen increased the odds of a positive environment by a factor of 2.4 and the average amount of MAP was increased by 3.5 cfu/g. We were not able to model the relationship between non-pen environmental sample status and the distance between shedding animals and the sample's location, and neighboring pens did not significantly affect the results of the pen-level analysis. The amount of MAP in pen-level samples and the probability of a pen testing positive for MAP were both positively but non-significantly correlated with the number of animals in the pen shedding >30 cfu/g of MAP. At least 6 environmental samples met the criteria for the U.S. Voluntary Bovine Johne's Disease Control Program on 47 of the 72 sampling dates; of these, 19 of the 47 FC-positive sampling dates were positive by the 6-sample environmental testing method, resulting in a herd sensitivity of 0.40 (95% CI: 0.26-0.54). None of the 3 FC-negative sampling dates produced positive environmental samples. Although environmental sampling can be used as a tool in understanding the level of MAP infection in a herd or pen, it did not appear to be a sensitive diagnostic method for herd positivity in these low prevalence herds, and its use may require caution.     

Risk management of paratuberculosis in dairy herds

Risk management of paratuberculosis (Johne's disease) in a dairy herd requires an assessment of the likelihood of paratuberculosis occurring in the herd, the economic impact of paratuberculosis on the herd and an evaluation of measures that can be taken to reduce this likelihood and impact.The likelihood of paratuberculosis occurring in the herd is related to the regional herd-level prevalence of paratuberculosis and the herd management (e.g., introducing animals from other herds). The economic impact of paratuberculosis includes production losses due to subclinical and clinical cases, losses due to increased replacement of animals and costs of control measures. Furthermore, a reduction of the price of milk from infected herds might result from consumer concerns about the zoonotic potential of paratuberculosis.Measures that reduce the likelihood of paratuberculosis occurring in a herd and its impact include preventive management measures (e.g., closed herd management and an effective separation of susceptible young stock from adult cattle), test-and-cull schemes for known infected herds and quality assurance schemes for test-negative herds. Quality assurance schemes for test-negative herds, such as schemes for 'low-Map bulk milk' and 'Map-free' herds, aim at safeguarding or increasing the profitability of these herds.Keys to success of risk management of paratuberculosis include realistic expectations of the results of paratuberculosis control, quality assurance and control programmes that are appreciated by farmers and incentives for farmers to participate in such programmes.     

Identification of Mycobacterium avium ssp. paratuberculosis infected dairy herds by environmental sampling

In herds with known prevalence (P) use of environmental sampling (ES) to detect Mycobacterium avium ssp. paratuberculosis (MAP) infected cattle herds was proofed in relation to P. In 31 MAP-infected free stall dairy herds and 15 non-infected herds P was defined by annually repeated whole herd testing by fecal culture (34 877 individual samples). Eight infected herds had a very low (> 0-2%), 14 a low (> 2-5%), four a medium (> 5-10%), and five a high P (> 10%). A mean number of nine environmental samples per herd were collected from the floor of lactating cows, milking, calving and sick cow areas and the crossover to the calf area. After twelve weeks cultivation on HEYM-medium with and without mycobactin positive samples were further characterized by PCR. All non-infected herds (100%) showed negative and 22 (71%) of the infected herds positive results in ES. Nine infected herds with negative ES results had a low P (0.04-4,04%). Proportion of positive ES depended on P and on sampling areas with 53.3% positive results in lactating cow areas and 45.2% in milking areas. For P > 5%, ES in these two areas caused a positive herd status; herds with P < 5% required sampling in the other areas too. The ES method has a herd sensitivity of 87% for dairy herds with P > 2% and provides an efficient tool to determine MAP infection status or herd prevalence.     

Prevalence of Salmonella in dairy herds in Alberta

Fifty dairy herds in Alberta were tested for the presence of Salmonella. Four (8%) dairy herds had at least 1 cow shedding Salmonella. Different isolates were identified by serotyping, phage typing, and antibiotic resistance patterns. Pulsed-field gel electrophoresis patterns were determined for unique isolates.     

Mycobacterium avium subsp. paratuberculosis from free-ranging deer and rabbits surrounding Minnesota dairy herds

The objectives of this study were to estimate the prevalence of Mycobacterium avium subsp. paratuberculosis (MAP) among deer and rabbits surrounding infected and noninfected Minnesota dairy farms using fecal culture, and to describe the frequency that farm management practices were used that could potentially lead to transmission of infection between these species. Fecal samples from cows and the cow environment were collected from 108 Minnesota dairy herds, and fecal pellets from free-ranging white-tailed deer and eastern cottontail rabbits were collected from locations surrounding 114 farms; all samples were tested using bacterial culture. In addition, a questionnaire was administered to 114 herd owners. Sixty-two percent of the dairy herds had at least 1 positive fecal pool or environmental sample. A total of 218 rabbit samples were collected from 90% of the herds, and 309 deer samples were collected from 47% of the herds. On 2 (4%) of the farms sampled, 1 deer fecal sample was MAP positive. Both farms had samples from the cow fecal pool and cow environment that were positive by culture. On 2 (2%) other farms, 1 rabbit fecal sample was positive by culture to MAP, with one of these farms having positive cow fecal pools and cow environmental samples. Pasture was used on 79% of the study farms as a grazing area for cattle, mainly for dry cows (75%) and bred or prebred heifers (87%). Of the 114 farms, 88 (77%) provided access to drylot for their cattle, mainly for milking cows (77/88; 88%) and bred heifers (87%). Of all study farms, 90 (79%) used some solid manure broadcasting on their crop fields. Of all 114 farms, the estimated probability of daily physical contact between cattle manure and deer or rabbits was 20% and 25%, respectively. Possible contact between cattle manure and deer or rabbits was estimated to occur primarily from March through December. The frequency of pasture or drylot use and manure spreading on crop fields may be important risk factors for transmission of MAP among dairy cattle, deer, and rabbits. Although the MAP prevalence among rabbits and deer is low, their role as MAP reservoirs should be considered.     

Risk factors associated with Mycobacterium avium subspecies paratuberculosis seropositivity in Canadian dairy cows and herds

Our objective was to determine the risk factors associated with the seroprevalence of Mycobacterium avium subspecies paratuberculosis (MAP) in a large number of randomly selected Canadian dairy herds, controlling for important confounding variables and co-infections with bovine leukemia virus (BLV), bovine viral diarrhea virus (BVDV) and Neospora caninum (NC). Serum samples from 30 randomly selected cows, where available, in 315 herds from seven provinces were tested for antibodies against BLV, MAP and NC using commercially available enzyme-linked immunosorbant assay (ELISA) test kits, while five unvaccinated cattle >6 months old from each herd were tested for antibodies to BVDV. We used a zero-inflated negative-binomial (ZINB) multivariable model to determine simultaneously the risk factors associated with the count of MAP-seropositive cows in a herd, and the odds of herds having no MAP-seropositive cows as compared to having one or more MAP seropositive cows in a herd. The following factors were significantly positively associated with the count of MAP-seropositive cows: "more than one cow in the maternity pen", "group-housing for pre-weaned calves in winter", "open heifers purchased during the last 12 months", "beef cattle direct (nose-to-nose) contact", "BVDV-seropositive herds (> or = 1 animal with > or = 1:64 titer)" and "BVD vaccination not done properly in calves" (i.e. after 6 months old, animals were not boostered 2-4 weeks after their first killed vaccine, or not given modified live vaccine), with count ratios of 1.7, 2.0, 2.3, 1.9, 1.4 and 1.8, respectively. The variable "BVDV vaccination (modified live) done properly in calves" (i.e. received another modified live vaccination after 6 months as well) was associated with 0.4 times fewer MAP-seropositive cows.     

Herd-level prevalence of Mycobacterium avium subsp. paratuberculosis infection in United States dairy herds in 2007

Testing of composite fecal (environmental) samples from high traffic areas in dairy herds has been shown to be a cost-effective and sensitive method for classification of herd status for Mycobacterium avium subsp. paratuberculosis (MAP). In the National Animal Health Monitoring System's (NAHMS) Dairy 2007 study, the apparent herd-level prevalence of MAP was 70.4% (369/524 had ≥1 culture-positive composite fecal samples out of 6 tested). Based on these data, the true herd-level prevalence (HP) of MAP infection was estimated using Bayesian methods adjusting for the herd sensitivity (HSe) and herd specificity (HSp) of the test method. The Bayesian prior for HSe of composite fecal cultures was based on data from the NAHMS Dairy 2002 study and the prior for HSp was based on expert opinion. The posterior median HP (base model) was 91.1% (95% probability interval, 81.6 to 99.3%) and estimates were most sensitive to the prior for HSe. The HP was higher than estimated from the NAHMS Dairy 1996 and 2002 studies but estimates are not directly comparable with those of prior NAHMS studies because of the different testing methods and criteria used for herd classification.     

Bacteriological investigations about paratuberculosis in dairy herds in Switzerland

In Switzerland clinical bovine paratuberculosis is registered sporadically with on average seven outbreaks per year. Our present studies are aimed to investigate the prevalence of Mycobacterium avium ssp. paratuberculosis (MAP)-infections in the Swiss cattle population and, therefore methods to culture MAP from bovine feces as well as a commercially available ELISA to detect MAP-specific antibodies are evaluated by using fecal samples and blood sera from herds with cases of clinical paratuberculosis. A series of molecular methods i.e. PCR-coupled RFLP analysis of the IS1311-insertion element of M. avium, PCR-coupled RFLP-analysis of the mycobacterial rpoB-gene, and DNA analysis of the mycobacterial 16S rRNA gene are used to identify mycobacterial isolates grown from bovine feces. Up to now, MAP was detected by culture in 12 of 155 (7.7%) animals from herds with paratuberculosis. A rather striking result is the finding of atypical mycobacteria in feces of 75 cattle (48.3%). Among these isolates, M. avium ssp. avium, M. thermoresistibile, and M. hassiacum/M. buckleii have been identified so far.     

The prevalence of milk and serum antibodies to Mycobacterium avium subspecies paratuberculosis in dairy herds in Ontario

This study randomly surveyed 50 dairy herds in Ontario; 18% and 30% of herds had 2 or more milk or serum enzyme-linked immunosorbent assay (ELISA)-positive cows, respectively. The apparent cow level prevalence was 1.7% and 2.6% on the milk and serum ELISA, respectively. The serum and milk assays agreed moderately.     

Impact of imperfect Mycobacterium avium subsp. paratuberculosis vaccines in dairy herds: A mathematical modeling approach

The objective of this study was to investigate the potential impacts of imperfect Mycobacterium avium subsp. paratuberculosis (MAP) vaccines on the dynamics of MAP infection in US dairy herds using a mathematical modeling approach. Vaccine-based control programs have been implemented to reduce the prevalence of MAP infection in some dairy herds; however, MAP vaccines are imperfect. Vaccines can provide partial protection for susceptible calves, reduce the infectiousness of animals shedding MAP, lengthen the latent period of infected animals, slow the progression from low shedding to high shedding in infectious animals, and reduce clinical disease. To quantitatively study the impacts of imperfect MAP vaccines, we developed a deterministic multi-group vaccination model and performed global sensitivity analyses. Our results explain why MAP vaccination might have a beneficial, negligible, or detrimental effect in the reduction of prevalence and show that vaccines that are beneficial to individual animals may not be useful for a herd-level control plan. The study suggests that high efficacy vaccines that are aimed at reducing the susceptibility of the host are the most effective in controlling MAP transmission. This work indicates that MAP vaccination should be integrated into a comprehensive control program that includes test-and-cull intervention and improved calf rearing management.     

Herd characteristics and management practices associated with seroprevalence of Mycobacterium avium subsp paratuberculosis infection in dairy herds

OBJECTIVE: To investigate herd characteristics and management practices associated with a high seroprevalence of Mycobacterium avium subsp paratuberculosis (MAP) in dairy herds in central California. SAMPLE POPULATION: 60 randomly selected cows from each of 21 dairy herds. PROCEDURES: Sera of selected cows were tested for antibodies against MAP by use of an ELISA test kit. Cows with a test sample-to-positive control sample (S:P) ratio of > or = 0.25 were considered seropositive, and herds with > or = 4% seropositive cows were considered high-seroprevalence herds. Data on herd characteristics and management practices were collected via interviews with owners. Bayesian logistic regression was used to model the predictive probability of a herd having a high seroprevalence on the basis of various herd characteristics and management practices. RESULTS: 9 of 21 (43%) herds were classified as high-seroprevalence herds. Five variables (history of previous signs of paratuberculosis in the herd, herd size, exposing cattle to water from manure storage lagoons, feeding unsalable milk to calves, and exposing heifers < or = 6 months old to manure of adult cows) were included in the predictive model on the basis of statistical and biological considerations. In large herds, the predictive probability of a high seroprevalence of MAP infection decreased from 0.74 to 0.39 when management changed from poor to good practices. In small herds, a similar decrease from 0.64 to 0.29 was predicted. CONCLUSIONS AND CLINICAL RELEVANCE: The seroprevalence of MAP infection in California dairies may be reduced by improvements in herd management practices.     

Estimate of the direct production losses in Canadian dairy herds with subclinical Mycobacterium avium subspecies paratuberculosis infection

The objective of this study was to estimate the annual losses from Mycobacterium avium subspecies paratuberculosis (MAP) for an average, MAP-seropositive, Canadian dairy herd. A partial-budget simulation model was developed with 4 components of direct production losses (decreased milk production, premature voluntary culling, mortality, and reproductive losses). Input values were obtained primarily from a national seroprevalence survey of 373 Canadian dairy farms in 8 of 10 provinces. The model took into account the variability and uncertainty of the required input values; consequently, it produced probability distributions of the estimated losses. For an average Canadian dairy herd with 12.7% of 61 cows seropositive for MAP, the mean loss was $2992 (95% C.I., $143 to $9741) annually, or $49 per cow per year. Additional culling, decreased milk production, mortality, and reproductive losses accounted for 46%, 9%, 16%, and 29% of the losses, respectively. Canadian dairy producers should use best management practices to reduce these substantial annual losses.     

Decision analysis model for paratuberculosis control in commercial dairy herds

A previous economic test-and-cull decision analysis model has been strengthened and updated with current epidemiologic information. Created using Excel and PrecisionTree software, the model incorporates costs and benefits of herd management changes, diagnostic testing, and different management actions based on test results to control paratuberculosis in commercial dairy herds. This novel "JD-Tree" model includes a herd management decision node (four options), a test/no test decision node (two options), a diagnostic test choice decision node (five options), test result chance nodes (four levels of possible results), and test action decision nodes (three options; cull, manage, no action). The model culminates in a chance node for true infection status. Outcomes are measured as a net cost-benefit value to the producer. The model demonstrates that improving herd management practices to control infection spread (hygiene) is often more cost-effective than testing; not all herds should test as part of a paratuberculosis control program. For many herds, low-cost tests are more useful than more sensitive, higher cost tests. The model also indicates that test-positive cows in early stages of infection may be retained in the herd to generate farm income, provided they are managed properly to limit infection transmission. JD-Tree is a useful instructional tool, helping veterinarians understand the complex interactions affecting the economics of paratuberculosis control and to define the accuracy and cost specifications of better diagnostic tests.     

Evaluation of cost-effectiveness of targeted sampling methods for detection of Mycobacterium avium subsp paratuberculosis infection in dairy herds

OBJECTIVE: To investigate the epidemiologic and financial impacts of targeted sampling of subpopulations of cows, compared with random sampling of all cows, for classification of dairy herd infection status for paratuberculosis. ANIMALS: All cows from 4 infected herds with a low-to-moderate prevalence of paratuberculosis and from 1 noninfected herd in California. PROCEDURE: The infection status of each cow was classified on the basis of results of an ELISA or combined ELISA and fecal culture results. Thirteen sampling schemes designed to randomly sample cows on the basis of lactation number, stage of lactation, and milk production were evaluated. Sampling without replacement was used to obtain a probability of herd detection of paratuberculosis for each evaluated sampling method and for simulated sample sizes between 30 and 150 cows. Marginal cost-effectiveness analysis was used to determine the cost increase relative to the increase in detection probability. RESULTS: Sampling cows in the third or higher lactation and > or = 200 days into lactation yielded the highest detection probability in most instances, resulting in a detection probability that was 1.4 to 2.5 times that obtained by sampling 30 cows in the second or higher lactation. Costs of testing via the alternative method with a 95% detection probability were approximately dollar 300 lower in a high-prevalence herd (31%) and dollar 800 lower in a low-prevalence herd (9%), compared with use of the reference method. CONCLUSIONS AND CLINICAL RELEVANCE: Detection of herds with paratuberculosis could be improved, and costs of testing substantially reduced by sampling targeted groups of cows.     

Bayesian kriging of seroprevalence to Mycobacterium avium subspecies paratuberculosis and Neospora caninum in Alberta beef and dairy cattle

Identifying spatial patterns of risk is important in the study of diseases with ecologic causes. Furthermore, relatively complex hierarchical modeling is required to determine how factors that are organized across levels interact, such as how an ecologic cause interacts with farm management and with animal characteristics. The objective of this study was to map the risk for Mycobacterium avium subspecies paratuberculosis (MAP - the causative agent of Johne's disease) and Neospora caninum (NC - the cause of neosporosis) infections in Alberta beef and dairy cattle. This objective utilized Bayesian generalized linear kriging to partition herd effects into a portion attributable to location and a portion that was independent of location. Seropositivity to NC in beef cattle showed strong support for spatial covariance, suggesting that ecologic causes were important for beef cattle but not dairy cattle. There was little evidence of spatial covariance for MAP seropositivity in either beef or dairy cattle.     

Herd management practices associated with paratuberculosis seroprevalence in Dutch dairy herds

We describe the paratuberculosis management practices applied in dairy herds in the Netherlands. The findings from paratuberculosis seronegative and seropositive herds were compared to discover possible risk factors. In total, 370 randomly selected herds with > or =20 dairy cows were surveyed. A questionnaire was used to collect data on current and previous paratuberculosis management practices. All cattle aged > or =3 years were serologically tested for paratuberculosis using an enzyme-linked immunosorbent assay. Herds with >33 tested cattle, of which only one was seropositive, were excluded to reduce the risk of including false-positive herds in the analysis. A comparison of the management data of the seronegative herds (n = 166) and the seropositive herds (n = 143) showed that in both groups important management measures for the prevention of paratuberculosis, such as calving in a cleaned calving area, removing the calf immediately after birth, and feeding paratuberculosis non-suspect roughage to calves, were used only rarely. However, such measures should be regarded as the critical first step to control the disease and/or reduce its prevalence. Using univariable analysis, four factors were statistically different between seronegative and seropositive herds: herd size, cows with clinical signs of paratuberculosis, prompt selling of clinically diseased cattle and feeding milk replacer. Using a multivariable logistic regression model, only herd size was a significantly different factor. These results indicate that most of the paratuberculosis preventive management measures were executed on these Dutch dairy farms only to a limited extent.     

Observations on the control and eradication of paratuberculosis in dairy herds

Johne's disease is widely seen in dairy herds in Germany. Estimates based primarily on epidemiological surveys in neighbouring states assume that 5 to 15 % of German herds are infected. In the past three years several authors have reported that the causative agent of Johne's disease, Mycobacterium avium subspecies paratuberculosis (MAP), is found ubiquitously in the environment and can be isolated from a number of different animals, including non ruminants. These results imply that MAP should be considered an environmental pathogen. Based on this assumption a concept for control and eradication of Johne's disease is presented aiming at minimizing the future spread of disease and reducing environmental contamination with the pathogen at low costs. The concept includes the classification of herds based on an bulk milk ELISA followed by a robot-compatible bulk milk PCR in ELISA-positive herds only. Due to the comparatively low costs combined with the high specificity of the approach a detection of heavily infected herds ("tip of the iceberg") all over the country would be possible; based on the eradication of strong shedders in these herds the input of MAP into the environment would be reduced considerably.     

Evaluation of bacteriologic culture of individual and pooled fecal samples for detection of Mycobacterium paratuberculosis in dairy cattle herds

OBJECTIVES: 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. STUDY DESIGN: Cross-sectional study. ANIMALS: 24 dairy cattle herds. PROCEDURE: Individual and pooled fecal samples were submitted for bacteriologic culture, and results were compared between these groups. RESULTS: Ninety-four and 88% of pooled fecal samples that contained feces from at least 1 animal with high (mean, > or = 50 colonies/tube) and moderate (mean, 10 to 49 colonies/tube) concentrations of M paratuberculosis, respectively, were identified by use of bacteriologic culture of pooled fecal samples. Prevalences of paratuberculosis determined by bacteriologic culture of pooled and individual fecal samples were highly correlated. CONCLUSIONS AND CLINICAL RELEVANCE: Bacteriologic culture of pooled fecal samples provided a valid and cost-effective method for the detection of M paratuberculosis infection in dairy cattle herds and can be used to estimate prevalence of infection within a herd.     

Evaluation of microbial culture of pooled fecal samples for detection of Mycobacterium avium subsp paratuberculosis in large dairy herds

OBJECTIVE: To evaluate sensitivity of microbial culture of pooled fecal samples for detection of Mycobacterium avium subsp paratuberculosis (MAP) in large dairy herds and assess the use of the method for estimation of MAP prevalence. ANIMALS: 1,740 lactating cows from 29 dairy herds in California. PROCEDURE: Serum from each cow was tested by use of a commercial ELISA kit. Individual fecal samples were cultured and used to create pooled fecal samples (10 randomly selected fecal samples/pool; 6 pooled samples/herd). Sensitivity of MAP detection was compared between Herrold's egg yolk (HEY) agar and a new liquid culture method. Bayesian methods were used to estimate true prevalence of MAP-infected cows and herd sensitivity. RESULTS: Estimated sensitivity for pooled fecal samples among all herds was 0.69 (25 culture-positive pools/36 pools that were MAP positive). Sensitivity increased as the number of culture-positive samples in a pool increased. The HEY agar method detected more infected cows than the liquid culture method but had lower sensitivity for pooled fecal samples. Prevalence of MAP-infected cows was estimated to be 4% (95% probability interval, 2% to 6%) on the basis of culture of pooled fecal samples. Herd-level sensitivity estimate ranged from 90% to 100% and was dependent on prevalence in the population and the sensitivity for culture of pooled fecal samples. CONCLUSIONS AND CLINICAL RELEVANCE: Use of pooled fecal samples from 10 cows was a cost-effective tool for herd screening and may provide a good estimate of the percentage of MAP-infected cows in dairy herds with a low prevalence of MAP.