Increased incidence rate of undesired early heifer departure in Mycoplasma bovis-antibody positive Danish dairy cattle herds.

Mycoplasma bovis infections cause disease and production losses in cattle worldwide. The long-term consequences are not well described despite being important for management decisions during and after disease outbreaks. We investigated the association between M. bovis antibody-positivity and undesired early departure (UED, i.e. death, euthanasia or slaughter) before first calving in a cohort of 636 heifers from 36 Danish dairy herds with and without a history of M. bovis-associated disease. The herds were visited 4 times at 3-month intervals and blood samples from young stock and milk samples from lactating cows were collected. Poisson regression was performed to examine the association with UED as outcome, logarithmic transformation of risk time as offset and herd as a random effect. Individual antibody measurements and group-level variables representing the infection level among young stock and cows, age and mortality variables were included in the model. The incidence rate ratio of UED increased by 1.23 times for every 10% increase in M. bovis young stock seroprevalence, while the effect of individual antibody level was modified by age and influenced UED less. In conclusion, UED in heifers was associated with M. bovis antibody-positivity in young stock and should be controlled in dairy herds to reduce losses.

Factors associated with variation in bulk tank milk Mycoplasma bovis antibody-ELISA results in dairy herds.

The relevance and limitations for using measurements of antibodies against Mycoplasma bovis in bulk tank milk (BTM) as a potentially cost-effective diagnostic tool for herd classification has not been evaluated before. Assuming that an increasing or high seroprevalence is a result of on-going or recent spread of M. bovis in a dairy herd, we tested the hypothesis that increasing prevalence of antibody-positive cows and young stock are associated with increasing BTM antibody ELISA values against M. bovis in Danish dairy herds with different courses of M. bovis infection. Furthermore, we tested whether herd size was associated with variations in the BTM responses. Thirty-nine Danish dairy herds selected to represent 4 different herd-level infection groups [8 control herds, 14 acute outbreak herds, 7 herds with previous outbreaks, and 10 herds with elevated BTM ELISA-values directed against M. bovis (>64% optical density measurement)] were visited 4 to 5 times, approximately 3mo apart. At each visit, 65 young stock were blood sampled. At the milk recording date closest to the herd visit date, 50 milk recording samples from individual lactating cows were randomly selected. In addition, a BTM sample was collected as a representative sample directly from the bulk tank by the dairies' milk truck drivers as part of the mandatory milk quality-control scheme. Blood and milk samples were tested for antibodies against M. bovis with a commercially available ELISA test (Bio-X BIO K 302, Bio-X Diagnostics, Rochefort, Belgium). A linear mixed effects model was used to analyze the effects of the prevalence of antibody-positive lactating cows and young stock and herd size on the BTM M. bovis ELISA results. Herd was included as a random effect to account for clustering of BTM samples originating from the same herd. Increasing prevalence of antibody-positive lactating cows was the only variable associated with increasing M. bovis BTM ELISA optical density measurement. In contrast, the prevalence of antibody-positive young stock did not correlate with the BTM optical density measurement. In conclusion, some M. bovis associated herd infections are detectable by BTM ELISA-testing, but limitations exist and further investigations of the effect of different clinical disease expressions in the herds are warranted.

Assessment of the probability of introducing Mycobacterium tuberculosis into Danish cattle herds.

Tuberculosis is a zoonosis caused by Mycobacterium spp. International trade in cattle is regulated with respect to Mycobacterium bovis (M. bovis) but not Mycobacterium tuberculosis (M. tuberculosis), despite that cattle can become infected with both species. In this study we estimated the annual probability (PIntro) of introducing M. tuberculosis into the Danish cattle population, by the import of cattle and/or by immigrants working in Danish cattle herds. Data from 2013 with date, number, and origin of imported live cattle were obtained from the Danish cattle database. Information on immigrants working in Danish cattle herds was obtained through a questionnaire sent to Danish cattle farmers. The gained inputs were fed into three stochastic scenario trees to assess the PIntro for the current and alternative test-and-manage strategies, such as testing of imported animals and/or testing immigrant workers with the tuberculin skin test. We considered the population of Danish farmers and practitioners free of tuberculosis, because in Denmark, the incidence of the disease in humans is low and primarily related to immigrants and socially disadvantaged people. The median annual probability of introducing M. tuberculosis into the Danish cattle population due to imported live cattle was 0.008% (90% P.I.: 0.0007%; 0.03%), while the probability due to immigrant workers was 4.1% (90% P.I.: 0.8%; 12.1%). The median combined probability (PIntro) due to imported cattle plus workers was 4.1% (90% P.I.: 0.8%; 12.6%). Hence, on average at least one introduction each 24 (90% P.I.: 8; 125) years could be expected. Imported live cattle appeared to play a marginal role on the overall annual PIntro, because they represented only approximately 0.2% of the median annual probability. By testing immigrant workers the overall annual PIntro could be reduced to 0.2% (90% P.I.: 0.04%; 0.7%). Thus, testing of immigrant workers could be considered as a risk mitigation strategy to markedly reduce the likelihood of introducing M. tuberculosis into the Danish cattle population, if the risk is considered unacceptable by the veterinary public health authorities.

Assessment of the probability of introduction of bovine tuberculosis to Danish cattle farms via imports of live cattle from abroad and immigrant workers.

Denmark has been recognized as officially free (OTF) from bovine tuberculosis (bTB) since 1980. In this study, we estimated the annual probability (PIntro) of introducing Mycobacterium bovis into the Danish cattle population, through (a) imports of cattle and (b) foreign personnel working in Danish cattle herds. Data from 2000 to 2013 with date, number and origin of imported live cattle were obtained from the Danish Cattle Federation. Information on immigrants working in Danish cattle herds was obtained through a questionnaire sent by email to a sample of Danish cattle farmers (N=460). Inputs obtained from data analysis, expert opinion, the questionnaire and literature were fed into three stochastic scenario tree models used to simulate the effect of import trade patterns, and contact between immigrant workers and cattle. We also investigated the opportunity of testing animals imported from OTF countries by tuberculin skin test and animals from non-OTF countries by interferon-γ test (IFN-γ), exemplified by using year 2009 where the number of imported animals was higher than usual. Results showed that PIntro is driven mainly by importation of live cattle. The combined median annual probability of introducing M. bovis into the Danish cattle population by either imported live cattle or infectious immigrant workers, ranged from 0.3% (90% prediction interval (P.I.): 0.04%:1.4%) in 2001 to 4.9% (90% P.I.: 0.6%; 19.2%) in 2009. The median of the median PIntro estimates from the 14 years was 0.7% (median of 90% P.I.: 0.08%; 3.5%). Hence, on average, at least one introduction each 143 years could be expected, if the annual number of imported animals does not change remarkably in the future. If the number of imported animals increases, compared to the years we analyzed, additional testing of imported cattle might be considered. For example, in 2009, PIntro would have been reduced from 4.9% to 0.8% (90% P.I.: 0.1%; 4.7%) if animals from OTF countries had been tested with the tuberculin skin test and animals from non-OTF countries had been tested with the IFN-γ test. The presented model could be used easily in other countries with similar bTB status to Denmark, where wildlife represents a negligible probability of infection for domestic cattle and where the imported live cattle represent the main pathway of bTB introduction into the local cattle population.

Challenges for bovine viral diarrhoea virus antibody detection in bulk milk by antibody enzyme-linked immunosorbent assays due to changes in milk production levels.

BACKGROUND: Bovine viral diarrhoea (BVD) is considered eradicated from Denmark. Currently, very few (if any) Danish cattle herds could be infected with BVD virus (BVDV). The Danish antibody blocking enzyme-linked immunosorbent assay (ELISA) has been successfully used during the Danish BVD eradication program, initiated in 1994. During the last decade, the cattle herd size has increased while the prevalence of BVDV has decreased. In this study, we investigated how these changes could affect the performance of the Danish blocking ELISA and of the SVANOVIR® BVDV-Ab indirect ELISA. The latter has successfully been used to eradicate BVD in Sweden. Data (2003-2010) on changes in median herd size and milk production levels, occurrence of viremic animals and bulk milk surveillance were analysed. Additionally, the Danish blocking ELISA and the SVANOVIR ELISA were compared analyzing milk and serum samples. The prevalence of antibody positive milking cows that could be detected by each test was estimated, by diluting positive individual milk samples and making artificial milk pools. RESULTS: During the study period, the median herd size increased from 74 (2003) to 127 cows (2010), while the prevalence of BVDV infected herds decreased from 0.51 to 0.02 %. The daily milk yield contribution of a single seropositive cow to the entire daily bulk milk was reduced from 1.61 % in 2003 to 0.95 % in 2010 due to the increased herd size. It was observed that antibody levels in bulk milk decreased at national level. Moreover, we found that when testing bulk milk, the SVANOVIR® BVDV-Ab can detect a lower prevalence of seropositive lactating cows, compared to the Danish blocking ELISA (0.78 % vs. 50 %). Values in the SVANOVIR® BVDV-Ab better relate to low concentrations of antibody positive milk (R(2) = 94-98 %), than values in the blocking ELISA (R(2) = 23-75 %). For sera, the two ELISAs performed equally well. CONCLUSIONS: The SVANOVIR ELISA is recommended for analysis of bulk milk samples in the current Danish situation, since infected dairy herds e.g. due to import of infected cattle can be detected shortly after BVDV introduction, when only few lactating cows have seroconverted. In sera, the two ELISAs can be used interchangeably.

Stochastic simulation modeling to determine time to detect Bovine Viral Diarrhea antibodies in bulk tank milk.

A stochastic simulation model was developed to estimate the time from introduction of Bovine Viral Diarrhea Virus (BVDV) in a herd to detection of antibodies in bulk tank milk (BTM) samples using three ELISAs. We assumed that antibodies could be detected, after a fixed threshold prevalence of seroconverted milking cows was reached in the herd. Different thresholds were set for each ELISA, according to previous studies. For each test, antibody detection was simulated in small (70 cows), medium (150 cows) and large (320 cows) herds. The assays included were: (1) the Danish blocking ELISA, (2) the SVANOVIR(®)BVDV-Ab ELISA, and (3) the ELISA BVD/MD p80 Institute Pourquier. The validation of the model was mainly carried out by comparing the predicted incidence of persistently infected (PI) calves and the predicted detection time, with records from a BVD infected herd. Results showed that the SVANOVIR, which was the most efficient ELISA, could detect antibodies in the BTM of a large herd 280 days (95% prediction interval: 218; 568) after a transiently infected (TI) milking cow has been introduced into the herd. The estimated time to detection after introduction of one PI calf was 111 days (44; 605). With SVANOVIR ELISA the incidence of PIs and dead born calves could be limited and the impact of the disease on the animal welfare and income of farmers (before detection) could be minimized. The results from the simulation modeling can be used to improve the current Danish BVD surveillance program in detecting early infected herds.

Quantitative assessment of the risk of introduction of bovine viral diarrhea virus in Danish dairy herds.

A quantitative risk assessment was carried out to estimate the likelihood of introducing bovine viral diarrhea virus (BVDV) in Danish dairy herds per year and per trimester, respectively. The present study gives important information on the impact of risk mitigation measures and sources of uncertainty due to lack of data. As suggested in the Agreement on the Application of Sanitary and Phytosanitary Measures (SPS Agreement), the OIE Terrestrial Animal Health Code was followed for a transparent science-based risk assessment. Data from 2010 on imports of live cattle, semen, and embryos, exports of live cattle, as well as use of vaccines were analyzed. Information regarding the application of biosecurity measures, by veterinarians and hoof trimmers practicing in Denmark and in other countries, was obtained by contacting several stakeholders, public institutions and experts. Stochastic scenario trees were made to evaluate the importance of the various BVDV introduction routes. With the current surveillance system, the risk of BVDV introduction was estimated to one or more introductions within a median of nine years (3-59). However, if all imported animals were tested and hoof trimmers always disinfected the tools used abroad, the risk could be reduced to one or more introductions within 33 years (8-200). Results of this study can be used to improve measures of BVD surveillance and prophylaxis in Danish dairy herds.