Predicting Positive ELISA Results in Dairy Herds with a Preferred Status in a Paratuberculosis Control Program.

Dairy herds participating in the Dutch milk quality assurance program for paratuberculosis are assigned a herd status on the basis of herd examinations by ELISA of individual serum or milk samples, followed by an optional confirmatory fecal PCR. Test-negative herds are assigned Status A; the surveillance of these herds consists of biennial herd examinations. Farmers falsely believing that their Status A herds are Map-free may inadvertently refrain from preventive measures. Therefore, we aimed to develop a predictive model to alert Status A farmers at increased risk of future positive ELISA results. Using data of 8566 dairy herds with Status A in January 2016, two logistic regression models were built, with the probabilities of ≥1 or ≥2 positive samples from January 2017-June 2019 as dependent variables, and province, soil type, herd size, proportion of cattle born elsewhere, time since previous positive ELISA results, and the 95th percentile of the S/P ratios in 2015-2016, as explanatory variables. As internal validation, both models were applied to predict positive ELISA results from January 2019-June 2021, in 8026 herds with Status A in January 2019. The model predicting ≥1 positive sample had an area under the receiver operating characteristics curve of 0.76 (95% CI: 0.75, 0.77). At a cut-off predicted probability πc = 0.40, 25% of Status A herds would be alerted with positive and negative predictive values of 0.52 and 0.83, respectively. The model predicting ≥2 positive samples had lower positive, but higher negative, predictive values. This study indicates that discrimination of Status A herds with high and low risks of future positive ELISA results is feasible. This might stimulate farmers with the highest risks to take additional measures to control any undetected Map infections.

Quantification of vertical and horizontal transmission of Neospora caninum infection in Dutch dairy herds.

Ninety-six of 108 randomly selected Dutch dairy herds had one or more cows with a positive serostatus for N. caninum. In these 96 herds, we have quantified the probabilities of vertical transmission (VT) and horizontal transmission (HT) of N. caninum infection by combining serostatus and pedigree data in 4091 dam-daughter pairs. The probability of animals infected by vertical transmission during pregnancy (Prob(VT)) was calculated as the proportion of seropositive daughters among daughters of seropositive dams. The probability of animals infected by horizontal transmission (Prob(HT)) was the proportion of seropositive daughters among daughters of seronegative dams. These probabilities were calculated after the frequencies of observed dam-daughter combinations were corrected for (1) imperfect test-characteristics, (2) underestimation of horizontal transmission in situations that seronegative dams were horizontally infected after the birth of their daughters and (3) overestimation of vertical transmission in situations that seronegative daughters born from seropositive dams were horizontally infected. The incidence rate for horizontal transmission was calculated based on Prob(HT) and the average age of the animals in these herds. Based on the analysis of dam-daughter serology, Prob(VT) was 61.8% (95% CI: 57.5-66.0%) and Prob(HT) was 3.3% (95% CI: 2.7-3.9%). After adjusting the observed frequencies for imperfect test-characteristics, underestimation of horizontal transmission and overestimation of vertical transmission, Prob(VT) decreased to 44.9% (95% CI: 40.0-49.9%) while Prob(HT) increased to 4.5% (95% CI: 3.9-5.2%). Prob(HT) corresponded with an incidence rate for horizontal transmission of 1.4 (95% CI: 1.2-1.7) infections per 100 cow-years at risk. When stratifying herds for the presence of farm dogs, Prob(HT) was higher (5.5% (95% CI: 4.6-6.4%)) in herds with farm dogs than in herds without farm dogs (2.3% (95% CI: 1.5-3.4%)). When stratifying for within-herd seroprevalence, Prob(HT) was higher (10.3% (95% CI: 8.6-12.2%)) in herds with high (> or =10%) within-herd seroprevalence compared with herds with low (<10%) within-herd seroprevalence (2.0% (95% CI: 1.5-2.6%)). Although there was this relation between Prob(HT) and within-herd seroprevalence (crude OR(PREV) = 5.7 (95% CI: 4.0-7.9)), in herds without farm dogs, this relationship was no longer statistical significant (OR(PREV|DOG-) = 1.9 (95% CI: 0.7-5.5)). It indicated that the association between seroprevalence and Prob(HT) depended largely on the presence of farm dogs. In addition, when looking for the presence of specific age-groups with significantly higher seroprevalence compared with the rest of the herd, there were 7 herds in which two or more horizontally-infected animals were present in specific age-groups. This was an indication of a recent point-source exposure to N. caninum. These results reiterate the current control strategies to apply strict dog-management measures as well as to minimize within-herd seroprevalence by monitoring serostatus of animals.

Factors associated with variation in Neospora caninum bulk-milk S/P ratios in initially bulk-milk negative testing Dutch dairy herds.

We conducted a study on 81 initially bulk-milk ELISA negative dairy herds taken from a random sample of Dutch dairy herds to evaluate variation in bulk-milk S/P ratios and to study reasons for bulk-milk conversion. These herds were repeatedly (3-month intervals) tested between April 2004 and August 2005 and serostatus of all animals had previously been established as negative (N), low-positive (LP) or high-positive (HP). Of these herds, herd- and test-related factors associated with variation in sample over positive (S/P) ratios were analysed using a multivariable linear-mixed model with 'herd' as random effect. In addition, changes of animal serostatus in converting herds were described. S/P ratios were calculated as the optical density of the bulk-milk sample minus the optical density of the negative serum control divided by the difference in optical density between the positive and negative serum control. Sixteen bulk-milk conversions in 12 dairy herds were seen with few indications of serological conversion in lactating cattle except for one herd in which recrudescence of infection appeared likely in nine cows. The effect of HP serostatus on bulk-milk S/P ratio was 2-3 times stronger compared with LP serostatus. In addition, bulk-milk S/P-ratio increased when the proportion of HP animals between 1 and 60 days in milk increased and decreased when the average milk-production level of the herd increased. Besides these herd-related factors, the use of different ELISA-testkits between test rounds had a significant effect on the S/P-ratio in bulk-milk samples.

Effect of Neospora caninum-serostatus on culling, reproductive performance and milk production in Dutch dairy herds with and without a history of Neospora caninum-associated abortion epidemics.

We quantified the effect of Neospora caninum (NC)-serostatus on culling and (re)production in 83 herds randomly selected from the Dutch dairy herd population (random group) and in 17 herds that had experienced an abortion epidemic associated with NC infection (epidemic-abortion group). In the random group, a single whole-herd blood sampling was done during the spring of 2003, while in the epidemic-abortion group whole-herd blood sampling was done repeatedly at least once a year starting after the abortion epidemic during the period 1997-2000 until the summer of 2004. Serological test-results for NC were given as 'negative' (N), 'low-positive' (LP) and 'high-positive' (HP). For analysing the time to culling, calving interval and age of first calving, survival analysis was used. For categorical reproduction parameters either a logistic-regression model (abortion, non-return after 1st insemination) or a Poisson-regression model (number of inseminations per pregnancy) was used. For milk production a linear-mixed model was used. All models were controlled, if applicable, for confounding variables like parity, production, season, year and abortion and adjusted for within-herd clustering. In random herds, HP serostatus increased the hazard for culling 1.73-fold (95% CI: 1.37-2.19) compared to N and LP serostatus. Compared to N serostatus, LP and HP serostatus in epidemic-abortion herds increased the odds for abortion 1.88-fold (95% CI: 1.41-2.52) and 1.72-fold (95% CI: 1.38-2.14), respectively. No other reproduction parameters were associated with NC-serostatus in the random or epidemic-abortion herds. We found no effect of serostatus on milk production in the random group. In contrast, milk production of LP and HP serostatus in the epidemic-abortion group was respectively, 0.72kgmilk/day (95% CI: 0.15-1.03) and 0.59kgmilk/day (95% CI: 0.13-1.30), less during the first 100 days of lactation in the first year after the abortion epidemic compared with N serostatus.