Environmental Survival of Mycobacterium avium subsp. paratuberculosis in Different Climatic Zones of Eastern Australia.

The duration of survival of both the S and C strains of Mycobacterium avium subsp. paratuberculosis in feces was quantified in contrasting climatic zones of New South Wales, Australia, and detailed environmental temperature data were collected. Known concentrations of S and C strains in feces placed on soil in polystyrene boxes were exposed to the environment with or without the provision of shade (70%) at Bathurst, Armidale, Condobolin, and Broken Hill, and subsamples taken every 2 weeks were cultured for the presence of M. avium subsp. paratuberculosis. The duration of survival ranged from a minimum of 1 week to a maximum of 16 weeks, and the provision of 70% shade was the most important factor in extending the survival time. The hazard of death for exposed compared to shaded samples was 20 and 9 times higher for the S and C strains, respectively. Site did not affect the survival of the C strain, but for the S strain, the hazard of death was 2.3 times higher at the two arid zone sites (Broken Hill and Condobolin) than at the two temperate zone sites (Bathurst and Armidale). Temperature measurements revealed maximum temperatures exceeding 60°C and large daily temperature ranges at the soil surface, particularly in exposed boxes.This work was supported by the NSW sheep industry through the NSW Ovine Johne's Disease Industry Fund and the NSW Rural Assistance Authority

Risk factors for ovine Johne's disease in infected sheep flocks in Australia

We conducted a cross-sectional study in 2004-05 to investigate risk factors for ovine Johne’s disease (OJD) involving 92 infected Merino sheep flocks in Australia. In each enrolled flock we collected pooled faecal-samples from 3- to 5-year-old sheep and cultured them for Mycobacterium avium subsp. paratuberculosis (MAP) to determine their OJD status. Based on pooled faecal-culture (PFC) results, three outcome variables representing different facets of disease biology were derived: pool OJD status (binomial: positive or negative), log pool MAP number (continuous) and cohort OJD prevalence level (ordinal: low (10%) prevalence). We used these outcomes in three separate multivariable analyses to identify risk factors, which were based on a questionnaire administered during a face-to-face interview with the farmer. We found higher OJD infection in sheep whose dams had been in poor condition and kept at a high stocking rate during lambing and in sheep which had experienced a longer period of growth retardation during their lifetime. Flocks that had vaccinated for >2 years (rather than only 1 to 2 years) with a killed MAP vaccine had significantly lower OJD infection. In addition, practices including culling low body-weight sheep or selling sub-flocks experiencing high losses, sharing of roads between neighbouring farms, and greater frequency of application of super-phosphate fertilizers were associated with higher OJD. Of the confounders investigated, infection was higher in flocks experiencing high mortalities; in wethers compared to ewes; and in 3-year-old sheep compared to 4-year-old sheep. Keywords: risk factors, paratuberculosis, ovine Johne’s disease, Australia, Mycobacterium, epidemiology, cross-sectional studyMeat and Livestock Australi

Association of farm soil characteristics with ovine Johne's disease in Australia

Speculation about the association of soil characteristics with the expression of ovine Johne’s disease (OJD) prompted this cross-sectional study. We enrolled 92 sheep flocks in Australia during 2004-05 and in each enrolled flock collected pooled faecal samples from an identified cohort (group of same age and sex) of sheep and soil samples from the paddocks grazed by this cohort of sheep. Faecal pools were cultured to create three outcome variables: positive or negative status of faecal pools (pool OJD status, binary); the log number of viable Mycobacterium avium subsp. paratuberculosis (MAP) organisms per gram of faeces (log pool MAP number, continuous); and the prevalence of faecal shedders (cohort OJD prevalence level, ordinal: low 10%). Separate statistical models were then developed to investigate the association between soil characteristics and each outcome variable. Sheep raised on soils with a higher percentage of organic carbon and clay had a higher OJD prevalence whereas, sheep grazing on soils with a higher content of sand and nitrogen had a lower OJD prevalence. Iron content of the soil was positively associated with OJD infection but the association between soil pH and OJD was inconclusive. Parent soil type, the only farm level factor, was not significant in any of the final models. Study results indicate a higher risk of OJD in sheep raised on soils with greater organic matter and clay content. We hypothesise that this is due to adsorption of MAP to clay and the consequent retention of the bacteria in the topsoil, thus making them available in higher numbers to grazing sheep. Keywords: risk factors, paratuberculosis, mycobacterium, cross-sectional study, organic carbon, clay, sand, pH, iron.Meat and Livestock Australi

Comparison of pre- and post-vaccination ovine Johne's disease prevalence using a Bayesian approach

postprintThis study was conducted to evaluate the effectiveness of GudairTM vaccine in decreasing the prevalence of shedding of Mycobacterium avium subsp. paratuberculosis (MAP) in flocks of varying initial prevalence. Thirty seven self-replacing Merino flocks from New South Wales and Victoria (Australia) that had been vaccinating lambs with GudairTM for at least five years were enrolled in the study. These flocks had been tested prior to or at commencement of vaccination using pooled faecal culture, agar gel immunodiffusion or both tests. These pre-vaccination test results were used to estimate pre-vaccination prevalence. Post-vaccination prevalence was estimated from culture of usually 7 pools of 50 sheep collected from the enrolled flocks in 2008-2009, approximately five or more years after commencement of vaccination. A Bayesian model was developed to estimate and compare the pre- and post-vaccination prevalences for the enrolled flocks. Apparent pre- and post-vaccination prevalences for flocks were modelled as functions of the true pre- and post-vaccination prevalences, respectively, and the sensitivities and specificities of the respective diagnostic tests. Logit-normal models were specified on pre- and post-vaccination true prevalences and were then used to make inferences about the median and 90th percentile of the prevalence distributions and their differences. Priors were mostly specified based on published literature or analysis of abattoir surveillance data for this population of flocks. The analysis found a significant decline in ovine Johne’s disease prevalence from a pre-vaccination median prevalence of 2.72% [95% probability interval (PI): 1.40; 6.86%] to a post-vaccination median prevalence of 0.72% (0.39; 1.27%). However 30 of the 37 flocks still contained sheep that were shedding MAP in their faeces. The results suggest that vaccination with Gudair™ is usually effective in reducing the prevalence of faecal shedding but the response to vaccination is variable among flocks. This approach could be implemented in similar situations to compare prevalences where information from multiple diagnostic tests with varied sensitivities and specificities is available. Keywords: Ovine Johne’s disease; Gudair; Vaccination; Abattoir surveillance; Faecal culture; Agar gel immune-diffusion test

Comparison of pre- and post-vaccination ovine Johne's disease prevalence using a Bayesian approach

This study was conducted to evaluate the effectiveness of GudairTM vaccine in decreasing the prevalence of shedding of Mycobacterium avium subsp. paratuberculosis (MAP) in flocks of varying initial prevalence. Thirty seven self-replacing Merino flocks from New South Wales and Victoria (Australia) that had been vaccinating lambs with GudairTM for at least five years were enrolled in the study. These flocks had been tested prior to or at commencement of vaccination using pooled faecal culture, agar gel immunodiffusion or both tests. These pre-vaccination test results were used to estimate pre-vaccination prevalence. Post-vaccination prevalence was estimated from culture of usually 7 pools of 50 sheep collected from the enrolled flocks in 2008-2009, approximately five or more years after commencement of vaccination. A Bayesian model was developed to estimate and compare the pre- and post-vaccination prevalences for the enrolled flocks. Apparent pre- and post-vaccination prevalences for flocks were modelled as functions of the true pre- and post-vaccination prevalences, respectively, and the sensitivities and specificities of the respective diagnostic tests. Logit-normal models were specified on pre- and post-vaccination true prevalences and were then used to make inferences about the median and 90th percentile of the prevalence distributions and their differences. Priors were mostly specified based on published literature or analysis of abattoir surveillance data for this population of flocks. The analysis found a significant decline in ovine Johne’s disease prevalence from a pre-vaccination median prevalence of 2.72% [95% probability interval (PI): 1.40; 6.86%] to a post-vaccination median prevalence of 0.72% (0.39; 1.27%). However 30 of the 37 flocks still contained sheep that were shedding MAP in their faeces. The results suggest that vaccination with Gudair™ is usually effective in reducing the prevalence of faecal shedding but the response to vaccination is variable among flocks. This approach could be implemented in similar situations to compare prevalences where information from multiple diagnostic tests with varied sensitivities and specificities is available. Keywords: Ovine Johne’s disease; Gudair; Vaccination; Abattoir surveillance; Faecal culture; Agar gel immune-diffusion test

Can early host responses to mycobacterial infection predict eventual disease outcomes?

Diagnostic tests used for Johne’s disease in sheep either have poor sensitivity and specificity or only detect disease in later stages of infection. Predicting which of the infected sheep are likely to become infectious later in life is currently not feasible and continues to be a major hindrance in disease control. We conducted this longitudinal study to investigate if a suite of diagnostic tests conducted in Mycobacterium avium subspecies paratuberculosis (MAP) exposed lambs at 4 months post infection can accurately predict their clinical status at 12 months post infection. We tracked cellular and humoral responses and quantity of MAP shedding for up to 12 months post challenge in 20 controls and 37 exposed sheep. Infection was defined at necropsy by tissue culture and disease spectrum by lesion type. Data were analysed using univariable and multivariable logistic regression models and a subset of variables from the earliest period post inoculation (4 months) was selected for predicting disease outcomes later on (12 months). Sensitivity and specificity of tests and their combinations in series and parallel were determined. Early elevation in faecal MAP DNA quantity and a lower interferon gamma (IFNγ) response were significantly associated with sheep becoming infectious as well as progressing to severe disease. Conversely, early low faecal MAP DNA and higher interleukin-10 responses were significantly associated with an exposed animal developing protective immunity. Combination of early elevated faecal MAP DNA or lower IFNγ response had the highest sensitivity (75%) and specificity (81%) for identifying sheep that would become infectious. Collectively, these results highlight the potential for combined test interpretation to aid in the early prediction of sheep susceptibility to MAP infection. KEYWORDS: Paratuberculosis; diagnostic tests; Mycobacterium; faecal DNA; Johne’s disease; interferon gamma.This work was supported by Meat and Livestock Australia and by Cattle Council of Australia, Sheepmeat Council of Australia and WoolProducers Australia through Animal Health Australia