Communication skills training and assessment of food animal production medicine veterinarians: A component of a voluntary Johne's disease control program.

In food animal production medicine (FAPM), the success of control programs for infectious diseases that have serious animal health and economic consequences frequently rely on the veterinarian's effective communication and producer adherence to veterinary recommendations. However, little research has been conducted on communication skills of practicing FAPM veterinarians. During this study, we developed a communication training workshop intervention to support the Atlantic Johne's Disease Initiative. Seventeen FAPM veterinarians across 10 clinics practicing within Maritime Canada participated in a pre-post intervention study design. Communication skills were evaluated utilizing 3 assessment tools; an objective structured clinical exam (OSCE), standardized client feedback, and an instrument designed for veterinary participants to assess their self-efficacy. Study results showed that before training, communication skills of participating veterinarians had limitations, including skill deficits in communication tasks strongly associated with increased adherence to veterinary recommendations. Based on the 3 assessment tools, communication skills of participating veterinarians improved with the training provided. Significant increases were detected in pre- to postintervention self-efficacy percentage scores, OSCE percentage and global scores from expert raters, and OSCE percentage and global scores from standardized client feedback. These improvements emphasize the importance of communication skills training specific to FAPM.

Dairy producer satisfaction and knowledge transfer with the veterinary-administered risk assessment and management plan in a voluntary Johne's disease control program.

The Atlantic Johne's Disease Initiative (AJDI) aims to control Mycobacterium avium ssp. paratuberculosis infection by using veterinary-administered risk assessments to identify high-risk management practices and prompt changes in management behavior. Objectives for this study were to measure producer satisfaction with the veterinary-administered risk assessment and management plan (RAMP) process in a voluntary Johne's disease (JD) control program, compare RAMP-specific satisfaction results based on herd JD status, and measure knowledge transfer from certified veterinarians to producers during the RAMP. A satisfaction questionnaire was adapted to the RAMP process in the AJDI to measure producer satisfaction. The questionnaire included 9 RAMP-specific producer satisfaction items, 1 global RAMP satisfaction item, and 16 questions to assess producer knowledge and knowledge translation about JD, bovine viral diarrhea (BVD), and bovine leukosis virus (BLV) during the RAMP (BVD and BLV used for comparison purposes). A total of 133 dairy producers in the AJDI (79.6% response rate) completed the questionnaire by telephone. The RAMP-specific satisfaction was high among the AJDI producers surveyed, and these results were not found to differ based on herd JD status. The lowest satisfaction scores and the highest number of "unable to assess" responses were for the item relating to cost. Factors that contributed to RAMP-specific producer satisfaction were not identified from the demographic and herd information available in this study. The knowledge scores indicated moderate knowledge about JD and fair knowledge about BVD and BLV. Evidence of knowledge translation from the RAMP was mixed in this study. Bovine viral diarrhea knowledge scores were not found to differ based on whether or not the certified veterinarian discussed BVD during the preceding RAMP, but BLV knowledge scores were higher among dairy producers that discussed BLV during the preceding RAMP. Strengths and gaps in producer knowledge about these 3 infectious diseases were identified. By using this producer questionnaire, interventions aimed at improving the content, delivery, and satisfaction of RAMP in JD control programs, such as the AJDI, can be developed.

Assessment of the relative sensitivity of milk ELISA for detection of Mycobacterium avium ssp. paratuberculosis infectious dairy cows.

Milk ELISA are commonly used for detection of Mycobacterium avium ssp. paratuberculosis (MAP) antibodies in dairy cows, due to low cost and quick processing for large numbers of samples. However, low sensitivity and variations from host and environmental factors can impede detection of MAP antibodies at early disease stages. The objectives of our study were to assess the sensitivity of milk ELISA in comparison with fecal tests and to evaluate how detectable antibody concentrations in milk vary with changes in fecal shedding of MAP, cow age, cow parity, days in milk, and time of year. To compare the sensitivity of a commercial milk ELISA with solid and broth fecal culture and with fecal real-time PCR, a longitudinal study was performed for the identification of MAP-infectious animals as determined by prior fecal testing for MAP shedding. In addition, associations between variation in milk MAP ELISA score and changes in fecal MAP shedding, host age, days in milk, and season were evaluated. Monthly milk and fecal samples were collected over 1 yr from 46 cows that were previously shedding MAP in their feces. Sensitivity of milk ELISA was 29.9% (95% CI: 24.8 to 35.1%), compared with 46.7% (40.7 to 52.7%) for fecal solid culture, 55.0% (49.3 to 60.7%) for fecal broth culture, and 78.4% (73.3 to 83.1%) for fecal direct real-time PCR. The effect of stage of lactation could not be separated from the effect of season, with increased milk ELISA scores at greater days in milk in winter. However, unpredictable monthly variations in results were observed among the 3 assays for individual cow testing, which highlights the importance of identifying patterns in pathogen and antibody detection over time in MAP-positive herds.

Genome-Wide Diversity and Phylogeography of Mycobacterium avium subsp. paratuberculosis in Canadian Dairy Cattle.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative bacterium of Johne's disease (JD) in ruminants. The control of JD in the dairy industry is challenging, but can be improved with a better understanding of the diversity and distribution of MAP subtypes. Previously established molecular typing techniques used to differentiate MAP have not been sufficiently discriminatory and/or reliable to accurately assess the population structure. In this study, the genetic diversity of 182 MAP isolates representing all Canadian provinces was compared to the known global diversity, using single nucleotide polymorphisms identified through whole genome sequencing. MAP isolates from Canada represented a subset of the known global diversity, as there were global isolates intermingled with Canadian isolates, as well as multiple global subtypes that were not found in Canada. One Type III and six "Bison type" isolates were found in Canada as well as one Type II subtype that represented 86% of all Canadian isolates. Rarefaction estimated larger subtype richness in Québec than in other Canadian provinces using a strict definition of MAP subtypes and lower subtype richness in the Atlantic region using a relaxed definition. Significant phylogeographic clustering was observed at the inter-provincial but not at the intra-provincial level, although most major clades were found in all provinces. The large number of shared subtypes among provinces suggests that cattle movement is a major driver of MAP transmission at the herd level, which is further supported by the lack of spatial clustering on an intra-provincial scale.

Novel Cell Preservation Technique to Extend Bovine In Vitro White Blood Cell Viability.

Although cell-mediated immunity based diagnostics can be integral assays for early detection of various diseases of dairy cows, processing of blood samples for these tests is time-sensitive, often within 24 hours of collection, to maintain white blood cell viability. Therefore, to improve utility and practicality of such assays, the objective of this study was to assess the use of a novel white blood cell preservation technology in whole bovine blood. Blood samples from ten healthy cows were each divided into an unpreserved control sample and a test sample preserved with commercially-available cell transport medium. Samples were maintained at room temperature and stimulated with the mitogens pokeweed and concanavalinA, as well as with interleukin-12 p40. Stimulation was completed on days 1, 5, and 8 post-sampling. Viability of white blood cells was assessed through interferon gamma production determined with a commercial enzyme linked immunosorbent assay. In addition, mononuclear cell viability was assessed with propidium iodide flow cytometry. Greater interferon gamma production was observed on days 5 and 8 post-collection in preserved samples, with both pokeweed and concanavalinA stimulating positive interferon gamma production on day 5 post-collection. A greater proportion of the amount of interferon gamma produced on day 1 continued to be produced on days 5 and 8 post-collection with concanavalinA stimulation (with or without interleukin 12) as compared to pokeweed stimulation. Additionally, viable mononuclear cells were still present at eight days post-collection, with a higher mean proportion detected at days 5 and 8 in all stimulated preserved samples. This practical and simple method to extend in vitro white blood cell viability could benefit the efficient utilization of cell-based blood tests in ruminants.

Limitations of variable number of tandem repeat typing identified through whole genome sequencing of Mycobacterium avium subsp. paratuberculosis on a national and herd level.

BACKGROUND: Mycobacterium avium subsp. paratuberculosis (MAP), the causative bacterium of Johne's disease in dairy cattle, is widespread in the Canadian dairy industry and has significant economic and animal welfare implications. An understanding of the population dynamics of MAP can be used to identify introduction events, improve control efforts and target transmission pathways, although this requires an adequate understanding of MAP diversity and distribution between herds and across the country. Whole genome sequencing (WGS) offers a detailed assessment of the SNP-level diversity and genetic relationship of isolates, whereas several molecular typing techniques used to investigate the molecular epidemiology of MAP, such as variable number of tandem repeat (VNTR) typing, target relatively unstable repetitive elements in the genome that may be too unpredictable to draw accurate conclusions. The objective of this study was to evaluate the diversity of bovine MAP isolates in Canadian dairy herds using WGS and then determine if VNTR typing can distinguish truly related and unrelated isolates. RESULTS: Phylogenetic analysis based on 3,039 SNPs identified through WGS of 124 MAP isolates identified eight genetically distinct subtypes in dairy herds from seven Canadian provinces, with the dominant type including over 80% of MAP isolates. VNTR typing of 527 MAP isolates identified 12 types, including "bison type" isolates, from seven different herds. At a national level, MAP isolates differed from each other by 1-2 to 239-240 SNPs, regardless of whether they belonged to the same or different VNTR types. A herd-level analysis of MAP isolates demonstrated that VNTR typing may both over-estimate and under-estimate the relatedness of MAP isolates found within a single herd. CONCLUSIONS: The presence of multiple MAP subtypes in Canada suggests multiple introductions into the country including what has now become one dominant type, an important finding for Johne's disease control. VNTR typing often failed to identify closely and distantly related isolates, limiting the applicability of using this typing scheme to study the molecular epidemiology of MAP at a national and herd-level.

Sensitivity and specificity of repeated test results from a commercial milk enzyme-linked immunosorbent assay for detection of Mycobacterium avium subspecies paratuberculosis in dairy cattle.

OBJECTIVE: To investigate the sensitivity and specificity of results of initial and repeated milk ELISAs (at 6- or 12-month intervals) to detect cows that were shedding Mycobacterium avium subsp paratuberculosis (ie, were infectious) and to evaluate factors influencing the probability that the results of a repeated milk ELISA would be positive for an infectious cow if the results of the initial milk ELISA were negative. DESIGN: Prospective cohort study. ANIMALS: 3,145 dairy cows from 32 herds. PROCEDURES: Herds from the 3 Maritime provinces in Canada (Prince Edward Island, New Brunswick, and Nova Scotia), participating in a Dairy Herd Improvement program, and that had undergone a prior Mycobacterium avium subsp paratuberculosis awareness project were selected for the study. Sample collection occurred between April 2009 and March 2011 with milk and fecal samples collected from all lactating cows in study herds every 6 months. Herds completing < 3 herd visits with collection of individual cow fecal or milk samples, within this sampling timeframe, were excluded from analyses. Fecal samples were cultured in liquid medium and a cow was defined as infectious if ≥ 1 sample was culture positive (reference test). A milk ELISA (index test) was completed with a commercial kit, following manufacturer's instructions. RESULTS: For a 6-month test interval, sensitivities of the milk ELISA to detect infectious cows were 22.0% and 32.6% for initial and combined initial and repeated tests (parallel interpretation), respectively. Specificity of the initial ELISA was 99.6% and was 99.2% for combined tests. For a 12-month test interval, sensitivities of the milk ELISA to detect infectious cows were 25.6% and 45.3% for initial and combined initial and repeated tests (parallel interpretation), respectively. Specificity of the initial ELISA was 99.6% and was 98.9% for combined tests. In infectious cows, magnitude of the initial negative ELISA result was a positive predictor for a positive repeated ELISA result. CONCLUSIONS AND CLINICAL RELEVANCE: Results of a repeated milk ELISA improved detection of Mycobacterium avium subsp paratuberculosis infectious cows, with minimal loss of specificity. A 12-month test interval provided a greater increase in sensitivity, relative to an initial test, than did a 6-month interval. Infectious cows with an initial negative milk ELISA result close to the cutoff for a positive test were more likely to have positive results on a repeated ELISA. Repeated testing improved detection of infectious cows and reduced risk of misclassification compared with a single ELISA result.

The association of detection method, season, and lactation stage on identification of fecal shedding in Mycobacterium avium ssp. paratuberculosis infectious dairy cows.

Mycobacterium avium ssp. paratuberculosis (MAP) is the causative organism of Johne's disease. Although fecal culture is considered the standard diagnostic test, the long incubation times, costs, and intermittent shedding of MAP hinder efficient screening programs based on culture results. The primary objectives of this study were to determine the detection ability of solid culture, broth culture, and real-time PCR (qPCR) for MAP in fecal samples and to assess how shedding patterns of MAP in feces vary with lactation stage and season. This knowledge could improve the use of these diagnostic assays in Johne's management programs. For this study, 51 MAP-infectious cows from 7 Atlantic Canadian dairy farms had fecal samples collected monthly over a 12-mo period. Samples were analyzed for MAP bacterial load via solid culture, broth culture, and qPCR. For all fecal samples, 46% [95% confidence interval (CI): 40 to 51%] were positive by solid culture, 55% (95% CI: 50 to 60%) by broth culture, and 78% (95% CI: 73 to 82%) by qPCR. Sensitivity of qPCR was numerically higher in the dry and postpartum lactation periods, and qPCR detection in summer and fall was 85% of that in winter and spring. Furthermore, culture-determined moderate or light shedding categories generally corresponded to qPCR cycle threshold values <35, but heavy shedding categories corresponded to qPCR values <29. Direct fecal qPCR is a MAP detection method that is quick and less costly than culture techniques, and it avoids the use of decontamination steps that could decrease numbers of bacteria in a sample below the detection limit. This study indicates that, for known MAP-positive cows, fecal qPCR had high sensitivity of MAP detection, thereby supporting the use of direct fecal qPCR as part of a Johne's herd control program.

Evaluation of environmental fecal culture for Mycobacterium avium subspecies paratuberculosis detection in dairy herds and association with apparent within-herd prevalence.

This study evaluated test characteristics of environmental culture (EC) for the detection of Mycobacterium avium subspecies paratuberculosis (MAP) in 32 herds over a 2-year period. Individual fecal samples were collected every 6 mo and environmental samples every 3 mo. Individual fecal culture was performed on samples from positive pools. Samples were cultured in broth, with confirmatory polymerase chain reaction performed on positive fecal samples. Repeated measures were accounted for using GEE logistic models. Relative to a MAP herd-status based on all pooled fecal culture results collected during the study, sensitivity of a set of 6 EC-samples collected from prescribed locations within the herd environment (EC-6) was 71% [95% confidence interval (CI): 49% to 86%] and specificity was 99% (95% CI: 95% to 100%). Sensitivity of EC increased as apparent within-herd fecal culture prevalence (aWHP) increased. The estimated aWHP increased as the proportion of positive EC-samples within an EC-6 set increased. Environmental culture is an acceptable tool for herd diagnosis of MAP in low-prevalence herds.

Évaluation de la culture fécale environnementale pour la détection deMycobacterium aviumsous-espèceparatuberculosisdans les troupeaux laitiers et l'association avec la prévalence apparente dans les troupeaux. Cette étude a évalué les caractéristiques des tests de cultures environnementales (CE) pour la détection de Mycobacterium avium sous-espèce paratuberculosis (MAP) dans 32 troupeaux pendant une période de deux ans. Des échantillons fécaux individuels ont été prélevés tous les 6 mois et des échantillons environnementaux tous les 3 mois. La culture fécale individuelle a été réalisée sur des échantillons provenant des échantillons regroupés positifs. Les échantillons ont été cultivés dans du bouillon et l'amplification en chaîne par la polymérase de confirmation a été réalisée sur des échantillons fécaux positifs. Des mesures répétées ont été enregistrées à l'aide de modèles logistiques d'équations généralisées d'estimation. En rapport avec un statut de troupeau pour MAP fondé sur tous les résultats des cultures fécales regroupées prélevées durant l'étude, la sensibilité d'un groupe de 6 échantillons-CE prélevés dans les lieux prescrits au sein de l'environnement du troupeau (CE-6) était de 71 % [intervalle de confiance (IC) de 95 %: de 49 % à 86 %] et la spécificité était de 99 % (IC de 95 %: de 95 % à 100 %). La sensibilité de CE a augmenté au fur et à mesure que la prévalence apparente de la culture fécale dans le troupeau (PaCF) montait. La PaCF estimée augmentait tandis que la proportion d'échantillons CE positifs dans le groupe CE-6 montait. La culture environnementale est un outil acceptable pour le diagnostic de MAP chez le troupeau pour les troupeaux avec une faible prévalence.(Traduit par Isabelle Vallières).

Comparison of bacterial culture, histopathology, and immunohistochemistry for the diagnosis of Johne's disease in culled dairy cows.

Paired samples of formalin-fixed, paraffin-embedded ileum and lymph node from 204 culled dairy cows were investigated for evidence of infection by Mycobacterium avium subsp. paratuberculosis. Of the samples, 151 were from animals that were tissue-culture positive for M. avium subsp. paratuberculosis, and 53 were from animals that were tissue and fecal culture negative. From the culture-positive animals, M. avium subsp. paratuberculosis was isolated from 78 samples of ileum and from 107 samples of lymph node. Ziehl-Neelsen acid-fast and immunoperoxidase stained slides were examined for 15 minutes each. Acid-fast organisms were identified in 7 of 78 (8.97%) and 6 of 106 (5.61%) culture-positive ileum and lymph node samples, respectively. Immunohistochemical (IHC) analysis of the same tissues identified infection in the ileum of 9 of 78 (11.54%) and in the lymph node of 5 of 106 (4.67%) culture-positive tissues. All tissues from culture-negative animals tested negative when using acid-fast and IHC staining. The sensitivity of these 2 tests in detecting M. avium subsp. paratuberculosis in culled dairy cows was not significantly different, and the tests exhibited substantial to almost perfect agreement. Both tests were much less sensitive than bacterial culture, detecting less than 6% of tissues positive compared with culture.

Johne's disease in Canada part II: disease impacts, risk factors, and control programs for dairy producers.

Part I of this 2-part review examined the clinical stages, pathophysiology, diagnosis, and epidemiology of Johne's disease, providing information relevant to Canada, where available. In Part II, a critical review of the economic impacts of the disease, risk factors, and important control measures are presented to enable Canadian bovine practitioners to successfully implement control strategies and participate in control programs. In cattle positive by enzyme-linked immunosorbant assay, there is a 2.4 times increase in the risk of their being culled, and their lactational 305-day milk production is decreased by at least 370 kg. Reduced slaughter value and premature culling account for losses of CDN dollars 1330 per year per infected 50-cow herd. Research has failed to show a consistent association between Mycobacterium avium subsp. paratuberculosis test status and reduced fertility or risk of clinical or subclinical mastitis. Host level factors include age and level of exposure, along with source of exposure, such as manure, colostrum, or milk. Agent factors involve the dose of infectious agent and strains of bacteria. Environmental management factors influence the persistence of the bacteria and the level of contamination in the environment. Emphasizing a risk factor approach, various control strategies are reviewed, including a number of national control programs currently in place throughout the world, specifically Australia, The Netherlands, and the United States. By reviewing the scientific literature about Johne's disease, control of the disease could be pursued through informed implementation of rational biosecurity efforts and the strategic use of testing and culling.

Johne's disease in Canada Part I: clinical symptoms, pathophysiology, diagnosis, and prevalence in dairy herds.

Recent international developments in the area of infectious disease control and nontariff trade barriers, along with possible zoonotic concerns, have provoked a revival of interest in Johne's disease in Canada and elsewhere. The bacterium causing Johne's disease, Mycobacterium avium subspecies paratuberculosis, is distributed worldwide and causes chronic granulomatous enteritis, also known as paratuberculosis, in domestic and exotic ruminants, including cattle. The subclinical form of this disease results in progressive weight loss, reduced milk production, lower slaughter value, and premature culling, with possible impacts on fertility and udder health. Eventually, infection can lead to the clinical form that manifests as chronic diarrhea, emaciation, debilitation, and eventual death. Currently, available tests to detect infected animals produce many false-negative results and some false-positives, particularly in subclinically infected animals, thus making their interpretation and utilization challenging in control programs. The objective of this 2-part review is to critically review the literature about Johne's disease in dairy cattle for bovine practitioners in Canada. Part I covers the clinical stages, pathophysiology, diagnosis, and prevalence of infection in Canada, while Part II discusses impacts, risk factors, and control programs relevant to Canadian dairy farms. By reviewing the scientific literature about Johne's disease, control of the disease could be pursued through informed implementation of rational biosecurity efforts and the strategic use of testing and culling.

Using and interpreting diagnostic tests.

Diagnostic tests are invaluable to the practice of veterinary medicine. Using them correctly and interpreting the results appropriately depend on having a good understanding of the basic principles outlined in this article. Topics covered include sensitivity and specificity, agreement among tests, using multiple tests, and other issues related to the use and interpretation of diagnostic tests. The most important principle is recognition that the interpretation of test results varies across populations and requires an estimate of the prevalence of the infection (or disease) in the population being studied.

Tetralogy of Fallot in a 2-year-old Holstein heifer.

A 2-year-old, purebred Holstein heifer with exercise intolerance and cardiovascular compromise was diagnosed at postmortem with tetralogy of Fallot, which typically results in death within a few months of life. Survival past the age of 2 was unexpected. The concurrent endocarditis of the pulmonic valve is discussed.