Herd-level occurrence and risk factors associated with respiratory and enteric pathogens from dairy calves in Ontario: A cross-sectional study.

This cross-sectional herd-level study aimed to determine the occurrence of and risk factors for pathogens associated with neonatal calf diarrhea and bovine respiratory disease on Ontario dairy farms. From April to August 2022, a convenience sample of 100 dairy farms were visited once. A questionnaire covering farm biosecurity, calving and colostrum management, preweaning nutrition, and housing was administered on-farm. At each farm visit, approximately 5 calves between 2 and 35 d old were randomly selected for fecal sampling. Furthermore, approximately 5 calves between 21 to 122 d old were randomly selected for nasopharyngeal sampling. In total, 363 fecal samples (from 83 dairy farms) and 390 nasopharyngeal swab samples (from 80 dairy farms) were collected. Fecal samples were analyzed individually using a multiplex PCR to identify bacterial and parasitic enteric pathogens. Nasopharyngeal swabs were analyzed as one pooled sample per farm using bacterial culture and real-time PCR. The most common enteric pathogens detected at herd-level were Cryptosporidium parvum (67.4%) and Escherichia coli K99+ (13.2%). The most common respiratory pathogens detected at herd-level were Pasteurella multocida (62.5%), bovine coronavirus (42.5%), and Mycoplasma bovis (21.2%). Multivariable logistic models were built to explore associations between the most common pathogens and herd-level predictors selected from the questionnaire. Herd positivity for C. parvum was positively associated with having more than 61 preweaned calves per year and feeding mainly whole milk to calves. The presence of M. bovis was positively associated with herds that combined manual and automatic milk-feeding systems, and the presence of bovine coronavirus was positively associated with having more than 98 preweaned calves during the year. Univariable Poisson regression models were built to explore the association between the most common pathogens and preweaning calf mortality. Herds that were positive for C. parvum, M. bovis, or bovine coronavirus had a greater risk of preweaning calf mortality. These results provide insights for future research on pathogens associated with NCD and BRD and offer guidance for veterinarians and dairy farmers in implementing disease control measures in dairy calf herds.

Using an automated tail movement sensor device to predict calving time in dairy cows.

This study aimed to evaluate the effectiveness of an automated tail movement sensor device (Moocall; Bluebell, Dublin, Ireland) to predict time of calving in dairy cows. At a commercial dairy farm in southern Ontario, Moocall (MC) devices were attached with the device's strap, and an additional elastic wrap, to the tail of cows approximately 3 d before their expected calving date. The MC has 2 types of alarm, a high activity alarm in the previous hour (1HA), and a high activity alarm in the previous 2 h (2HA); these alarms were sent and registered to the MC software. The calving and close-up pens were video monitored to determine the exact time of the onset of stage II of calving (amniotic sac visible at the vulva) and the end of stage II of calving (total expulsion of the calf). A total of 49 cows were enrolled, but we excluded 13 animals from the analysis as they had 3 or more MC drops from the tail (n = 6), a swollen tail (n = 3), or the MC device was lost (n = 4); this left 36 cows. In total, the device dropped off 21 (42%) cows. The average number of alarms (1HA and 2HA) per cow before stage II of calving was 2.7 ± 2.3 (± standard error). The first alarm after fitting the device on the tail was used to determine the device's sensitivity and specificity. Depending on the interval before the onset of parturition (i.e., 2, 4, 8, 12 h) in which the alarm was triggered, sensitivity varied from 5% to 72% and specificity from 50% to 93%. The false positive rate varied between 6% and 50% depending on the interval from the alarm to the onset of parturition. The high false positive and device drop rates (despite the addition of the elastic wrap) may compromise the applicability of this sensor device in a commercial setting.

Herd-level risk factors associated with preweaning mortality on Ontario dairy farms.

The objective of this cross-sectional, herd-level study was to identify associations between calf management practices and herd-level preweaning mortality on Ontario dairy farms. From April to August 2022, a convenience sample of 100 dairy farms from Ontario, Canada, were visited once. A questionnaire, covering farm biosecurity, calving, colostrum management, preweaning nutrition, and housing, was verbally administered at each farm visit. Furthermore, data regarding preweaning calf mortality were retrieved from each farm's herd management software or records, including the total count of calves that survived, were culled, or died during the preweaning period for the 365 d before the farm visit. Preweaning mortality was defined as the proportion of calves that died between ≥48 h and 60 d of age divided by the total number of calves alive after 48 h of age. The herd-level preweaning mortality risk on sampled farms ranged from 0 to 15.9%, with an average of 2.8% (SD = 3.8%). A multivariable Poisson regression model was used to evaluate associations between 22 explanatory variables and preweaning mortality. Factors associated with greater herd-level preweaning mortality were larger herd size; having treatment protocols for diarrhea, pneumonia or navel infection written in collaboration with a veterinarian (compared with farms with the same treatment protocols developed without a veterinarian), and the herd-veterinarian never inquiring about calf health (compared with farms where the herd veterinarian inquired sometimes). Factors associated with lower herd-level preweaning mortality were using the calving pen for sick cows, having more than 4 people working with calves, offering calves a minimum volume of ≥9 L of milk/day, and farmers with a level of formal education higher than secondary school. These results indicate that producers may be able to reduce preweaning calf mortality by providing adequate labor for calf care, offering calves sufficient volumes of milk, being proactive in communicating with their veterinary practitioners about calf health, and potentially by engaging in continuous education.

Exploring herd-level perinatal calf mortality risk factors in eastern Canadian dairy farms.

This closed cohort study aimed to identify the associations between dairy calf management practices and herd-level perinatal calf mortality risk. From February 2020 to June 2021, predominantly Holstein dairy farms in Québec (n = 1,832) and New Brunswick (n = 52), Canada, that were registered in the dairy herd improvement program were visited once. A questionnaire covering all aspects of precalving, calving, and colostrum management was administered. Data regarding perinatal mortality were retrieved from the dairy herd improvement program database for each farm for 2021. Perinatal mortality was calculated for each farm as the proportion of calves dead at birth or dying within 24 h after birth. A multivariable negative binomial model was used to assess herd-level factors associated with the risk of perinatal mortality. The final model included the lying surface in the calving area, the typical time to first colostrum intake, typical cow-calf contact time, the proportion of males born, the proportion of assisted calvings, and herd size. Herd-level perinatal mortality risk ranged from 0% to 38.1% (mean ± SE = 7.6% ± 0.1%). A greater proportion of males born, a higher proportion of assisted calvings, and delayed colostrum feeding were associated with increased herd-level perinatal mortality. Factors associated with a decreased herd-level perinatal mortality risk were having a typical cow-calf contact time between 7 and 12 h after calving compared with reduced cow-calf contact time, soft lying surfaces in the calving area compared with concrete and mat-lying surfaces, and an increased number of calvings per year. Our results show that although some of the significant risk factors are not well understood (i.e., calving area lying surface, typical cow-calf contact time), Canadian farmers could focus on the factors under their control (i.e., time to first colostrum feeding, proportion of difficult calvings, males born, and calvings per year) to reduce the risk of perinatal mortality. Future work should focus on qualitative research to understand the dairy farmer motivations and limitations to implementing practices identified in this and other studies to reduce perinatal mortality.

Bovine congenital defects recorded by veterinary practitioners.

A mobile phone app was used by 59 veterinary practitioners to collect case histories and images of 191 cattle with congenital defects distributed nationally over a 3-year period. The majority of cases were recorded during the spring calving season (57.6%) in pluriparous dairy dams. The majority of calves were recorded at birth or within the first week (66.5%) in singletons born at full-term. On the majority of farms (75.9%), this was the only congenitally deformed bovine recorded up to that point in the year and on the majority of farms, there were no congenitally deformed cattle recorded in the previous 5 years. The majority of congenital defects (83.5%) were recorded in the musculoskeletal or digestive systems. The three most commonly recorded individual defects were intestinal atresia (24.1%), schistosomus reflexus (20.4%) and ankylosis (6.8%); multiple defects were recorded in 13.1% of cases. These findings highlight the relatively high prevalence of intestinal atresia and schistosomus reflexus in calves attended by veterinary practitioners, which warrants implementation of preventive measures. The project highlights the potential benefits of veterinary-practitioner apps to detect changing trends in endemic, or the emergence of novel, congenital or other conditions.

A systematic review of the relationship between housing environmental factors and bovine respiratory disease in preweaned calves - Part 1: Ammonia, air microbial count, particulate matter and endotoxins.

Bovine respiratory disease (BRD) is one of the leading causes of mortality and morbidity in calves across diverse management systems. Despite expert opinion often citing the influence of housing environment on the level of respiratory disease in calf groups, there have been few reviews of environmental factors that predispose to BRD. This systematic review was undertaken to identify the measurable environmental variables associated with respiratory disease in housed preweaned calves. To achieve this Pubmed, CAB Direct and Scopus databases were searched. To be considered for inclusion, publications had to be fully published in English, published before 24 November, 2022 and include at least one measurable/ manipulated environmental variable and a standardized method of BRD detection. Twelve publications were included in this review. These examined a wide range of risk factors including air microbial count (four publications), air particulate matter (one publication); air endotoxins (one publication) and air ammonia (four publications). From the included publications, a statistically significant relationship to BRD was identified in 2/4 examining air microbial count, 1/1 examining air particulate matter, 1/1 examining air endotoxins and 2/4 examining air ammonia. This review indicated a paucity of evidence from the peer-review literature demonstrating a significant association between the many investigated exposure factors and BRD occurrence. An optimal environment for housed calves could not be clearly identified in this review.

A systematic review of the relationship between housing environmental factors and bovine respiratory disease in preweaned calves - Part 2: Temperature, relative humidity and bedding.

Bovine respiratory disease (BRD) is a challenge in all housed farming systems that raise calves. Farm to farm variation in BRD prevalence can be partially attributed to variation in host immunity, pathogens and housing environment. Unlike host immunity and BRD pathogens, housing environment has not been well investigated. The objective of this systematic review was to identify the measurable environmental variables associated with BRD in housed preweaned calves. Pubmed, CAB Direct and Scopus databases were searched. To be considered for inclusion publications had to be published in English, before 24 November, 2022 and include at least one measurable/ manipulated environmental variable and a standardized method of BRD detection. In total 12 publications were included in this review. In this second part of the systematic review the environmental variables identified were; temperature (9 publications); relative humidity (8 publications); bedding (5 publications); ventilation (1 publication); air CO2 concentration (1 publication) and air velocity (4 publications). Of the publications that were examined a statistically significant relationship to BRD was identified in 4/9 publications examining temperature, 3/8 examining relative humidity, 2/4 examining air velocity, 2/5 examining bedding, 0/1 examining ventilation rates and 0/1 examining CO2 concentration. From this review it is clear high airspeed at calf level should be avoided as should deep, wet pack bedding. The relationship between BRD prevalence and both high and low temperature requires more investigation to identify temperature thresholds associated with increased risk of BRD as well as the most influential modifiers. An optimal environment for housed calves could not be clearly identified in this review.

Review: Bovine foetal mortality - risk factors, causes, immune responses and immuno-prophylaxis.

This review of bovine foetal mortality (>42 d gestation) concluded that while the majority of risk factors associated with sporadic loss operate at animal-level, e.g. foetal plurality, those that operate at herd-level, e.g. some foetopathogenic infections, are more likely to result in abortion outbreaks. While the causes of foetal mortality have traditionally been classified as infectious and non-infectious, in fact, the latter category is a diagnosis of exclusion, generally without determination of the non-infectious cause. This review has also established that the traditional dichotomisation of infectious agents into primary and secondary pathogens is based on a flawed premise and these terms should be discontinued. The delicate balance of the maternal gestational immune system between not rejecting the allograft (conceptus) but rejecting (attacking) foetopathogens is stage-of-pregnancy-dependent thus the timing of infection determines the clinical outcome which may result in persistent infection or foetal mortality. Utilisation of our knowledge of the materno-foetal immune responses to foetopathogenic infection has resulted in the development of numerous mono- and polyvalent vaccines for metaphylactic or prophylactic control of bovine foetal mortality. While some of these have been shown to significantly contribute to reducing the risk of both infection and foetal mortality, others have insufficient, or conflicting evidence, on efficacy. However, recent developments in vaccinology, in particular the development of subunit vaccines and those that stimulate local genital tract immunity, show greater promise.

Impacts of dairy cow nutrition precalving on calf health.

This mini-review focuses on the effects of gestational dairy cow nutrition on calf health as mediated through colostrogenesis and calf immunity, morbidity, and mortality. The nutritional adequacy of the forage and supplementary diet and the metabolic status and body condition score of the dam can affect calf health. The mechanism of action of such impacts include maternal nutritional imbalances or deficiencies causing dyscolostrogenesis, nutritionally mediated calf ill health, and fetal programming impacts on calf health.

Invited review: Bovine neonatal morbidity and mortality-Causes, risk factors, incidences, sequelae and prevention.

The neonatal period may be defined as the first month of the calf's life; it may be considered part of the fourth trimester. It is the most hazardous due to the immaturity of the neonate's immune system and the environmental challenge from infections, the main causes of both bovine neonatal morbidity and mortality. The five most common morbidities causing mortality in neonatal calves are, in descending order, gastrointestinal infections, respiratory infections, abomasal disorders, umbilical infections and developmental abnormalities. This review describes the aetiology, incidence, risk factors and sequelae of these common morbidities and highlights current preventive strategies both at farm and national levels.

Growth rates of contract-reared versus home-reared replacement dairy heifers.

Successful heifer rearing is dependent on achieving optimal average daily gain (ADG) targets to calve for the first time at 24 months. Whilst dairy farmers internationally have traditionally managed their heifers on-farm to achieve these targets, recent dairy herd expansion within Europe has resulted in increased demand for labour-saving heifer-rearing strategies, such as off-farm contractrearing. However, loss of direct influence on the day-to-day management decisions affecting the ability of heifers to grow adequately to achieve this age of first calving may represent a potential barrier to uptake. Hence, the aim of this longitudinal observational study was to compare the growth rates of contract- vs home-reared heifers. Approximately 6 500 heifers from 120 commercial Irish dairy farms were enrolled in a 3-year study. For 65 of these farms, heifers were reared at a contract-rearing facility. For the remaining 55 farms, heifers were reared on their home farm. Over the course of 20 months from birth until precalving, heifers were examined and weighed at four farm visits. The relationship between several independent variables (farm type, herd size, heifer breed, economic breeding index (EBI) and health events) and ADG at different time points was investigated and analysed utilising linear mixed models. Overall ADG for heifers throughout the rearing period was 0.71 kg/day. There was a significant association between farm type and ADG for all five linear ADG models; home-reared heifers grew, on average, 0.025, 0.039, 0.11 and 0.059 kg/day more than contract-reared heifers between visit 1 and visit 4 (overall ADG), visit 1 and visit 2, visit 2 and visit 3 and visit 1 and visit 3, respectively. The occurrence of diarrhoea during farm visit 2 (median age 8.5 months) was associated with a significant reduction in ADG between visit 3 and visit 4. Calf-hood disease (diarrhoea, respiratory disease or navel ill) was not associated with the growth rate during any of the subsequent visit periods. While home-reared heifers had greater ADG during four of the five periods studied, median heifer ADG in both cohorts exceeded the minimum published target weight gains at each developmental stage required for heifers to reach puberty, conceive at 15 months and calve for the first time at 24 months. Importantly, there was wide variation both within enterprises and between farms. It was concluded that while the absolute difference in daily growth rates of home- and contract-reared heifers was minimal, when considered in the context of the entire heifer-rearing period, these growth rate differences have the potential to impact the future reproductive and milk production performance of heifers.

Characteristics (sensitivity and specificity) of herd-level diagnostic tests for Mycobacterium avium subspecies paratuberculosis in cattle - A systematic review.

Diagnostic tests for Johne's disease in cattle are characterised by poor sensitivity and often imperfect specificity at the animal level. Because farmers and veterinarians have limited assurance or confidence from results of testing individual animals for Mycobacterium avium subsp. paratuberculosis (MAP), control programmes based on whole herd-level testing provide the best framework for classifying herds. At the herd level, there is a diverse range of testing options for MAP based on both direct and indirect testing of individual and pooled samples. The most common measures of herd test performance, herd sensitivity (HSe) and herd specificity (HSp), are important for decision-making in herd test selection, for estimating prevalence and as inputs for simulation studies. This systematic review investigated the results of herd test evaluations for MAP in cattle, through a comprehensive search of the literature and a systematic four-stage screening process to identify relevant publications. Forty-six publications with relevant results were eligible for inclusion in the final review, containing evaluations of whole-herd ELISA serological testing, bulk milk tank ELISA, culture, PCR and phage testing, pooled faecal testing and environmental sample testing. Data extracted from each publication included sample populations, methods of analysis, reference tests, cut-off points, HSe and HSp. Direct comparisons between the reported HSe and HSp estimates of different studies is challenging due to the variations in herd prevalence and test protocols used. The data in this systematic review will benefit decision-makers and researchers and highlights knowledge gaps requiring further research.

A survey of biosecurity and health management practices on Irish dairy farms engaged in contract-rearing.

A survey was conducted to investigate potential differences in biosecurity and health management practices on Irish dairy farms that sent their heifers for contract-rearing (source dairy farms, SDF; n = 62) and those rearing their own heifers (control farms, CF; n = 50). Participating farmers were surveyed by postal questionnaire between September and November 2018. The overall response rate was 93%. Results show that structurally, SDF were larger, less fragmented, and more specialized than CF. Outsourcing of labor-intensive activities to external contractors was more common among SDF than CF, exposing them to potentially increased biosecurity risks associated with animal movements, use of shared equipment, and increased frequency of farm visitors. The majority of SDF sent heifers to a single-origin rearing facility (70%), with heifers most commonly arriving at the rearing unit between 2 and 4 mo (53%) and returning to the dairy farm between 18 and 21 mo of age (56%). Despite the increased biosecurity risk associated with contract-rearing, implementation of disease prevention measures was not superior on SDF compared with CF. For both farm types, there was scope for improvement to visitor biosecurity protocols, quarantine procedures, colostrum feeding practices, and hygiene of calving areas. This research provides an overview of the demographics and farm management practices implemented by dairy farmers engaged in contract-rearing of replacement heifers, and will serve to inform farmers, veterinary advisors, and policy makers.

The effect of dairy heifer pre-breeding growth rate on first lactation milk yield in spring-calving, pasture-based herds.

Optimising heifer growth rate may offer an opportunity to improve lifetime milk yield per cow, enhancing the environmental and economic efficiency of dairy farming operations. The effect of dairy heifer pre-breeding average daily weight gain (ADGPB) on first lactation milk yield was investigated. This observational study employed a data set comprising 265 Holstein-Friesian, or Holstein-Friesian-cross-Jersey heifers from seven commercial, spring-calving, pasture-based dairy herds, where the major component of the diet was grazed grass. These were weighed at birth and prior to breeding and ADGPB was calculated. Milk recordings were performed throughout the heifers' first lactation and 305-day yield figures calculated from these records. Yields were corrected to 4% fat and 3.1% protein to create standardised 305-day milk yield (SMY), which was the outcome of interest. Median ADGPB was 0.72 kg/day. Median 305-day yield was 5 967 kg. Linear regression was used to investigate the effect of weight and genetic, age and first calving factors on SMY. Pre-breeding average daily weight gain, age at first calving and predicted transmitting abilities for milk protein production and calving interval were all significant in the final model, which also included the random effects of farm and month of calving within year. ADGPB was quadratically related to first lactation SMY, with an ADGPB of 0.82 kg/day corresponding to the maximum predicted SMY. The model predicted that a heifer growing at 0.82 kg/day would produce 1 120 kg more SMY than a heifer growing at 0.55 kg/day, 218 kg more than a heifer growing at 0.7 kg/day and 103 kg more than a heifer growing at 0.90 kg/day. Manipulation of heifer growth rate may offer a viable method of increasing first lactation milk yield.

Assessing whether dairy cow welfare is "better" in pasture-based than in confinement-based management systems.

Consumers perceive pasture-based systems of milk production as natural and therefore better for cow welfare than confinement systems. However both systems are heterogeneous and continually evolving, varying from total confinement to total pasture with many hybrid intermediaries. To compare the welfare of dairy cows in these various systems, we use the three spheres framework, comprising biological functioning, natural behaviour and affective states. Considering biological functioning, pasture-based cows are less at risk of subclinical and clinical mastitis, claw lesions, lameness, metritis, early embryonic mortality, culling and mortality, but at more risk of internal parasitism, malnutrition and delayed onset of oestrous activity postpartum than confined cows. Regarding natural behaviours, pasture-based cows exhibit less agonistic behaviour, better lying behaviour, more normal oestrous behaviours and better synchronicity of behaviours than confined cows. They also have the opportunity to graze, which is one of the main features of the behavioural repertoire of dairy cows, but, they may also experience long periods away from pasture in larger herds, and severe climatic stresses which will become increasingly important as the climate changes. Our current ability to assess the affective state of dairy cows is poor. For example, hunger is an important subjective state that cannot be measured directly. The growing focus on ensuring that animals have lives worth living, means that dairy cows should garner some positive emotions from their lives, and it seems clear that pasture access is essential for this. Clearly measurement of affective state is an important challenge for future dairy cow welfare research. At the extremes of management systems, there can be major differences in animal welfare but in hybrid systems, dairy cows experience elements of both confinement and pasture which may ameliorate the negative effects of each on cow welfare. Ultimately, the optimal system gives cows an element of choice between both environments. Moreover management of the system, whether it is confinement or pastured-based, may be as important as the system of management in ensuring good dairy cow welfare and addressing societal concerns.Abbreviations: BCS: Body condition score; TMR: Total mixed ration.

The effect of growth rate on reproductive outcomes in replacement dairy heifers in seasonally calving, pasture-based systems.

The effect of average daily gain (ADG) on reproductive outcomes in replacement dairy heifers was investigated. All heifers were managed in the typical Irish spring calving, pasture-based system, where the herd calves in 1 block between January and April and the majority of the diet comprises grazed grass. Heifer calves (n = 399) from 7 herds were weighed at birth and at the beginning of the breeding season, and ADG was calculated. Service dates and pregnancy diagnosis results were recorded, and conception dates were calculated. Days open (DO) was defined as the number of days between the beginning of the breeding season and conception. Genetic data were retrieved from the Irish Cattle Breeding Federation database. A Cox proportional hazard model was constructed to identify variables with a significant effect on DO. An accelerated failure time model was used to predict survival curves and median survival times for different combinations of the significant variables. The ADG ranged from 0.41 to 0.91 kg/d, with a median of 0.70 kg/d. Frailty effect of farm within year, maintenance subindex of the economic breeding index, and ADG had a significant effect on DO. Derived from the final accelerated failure time model, the predicted median DO for a heifer with an ADG of 0.40, 0.70, or 0.90 kg/d aged 443 d at the beginning of the breeding season and with a maintenance subindex in the second tercile were 27, 16, and 11 d, respectively.

Sensitivity, specificity and predictive probability values of serum agglutination test titres for the diagnosis of Salmonella Dublin culture-positive bovine abortion and stillbirth.

The objective of this study was to determine the diagnostic value of maternal serology for the diagnosis of Salmonella Dublin bovine abortion and stillbirth. A retrospective, unmatched, case-control study was carried out using twenty year's data (1989-2009) from bovine foetal submissions to an Irish government veterinary laboratory. Cases (n = 214) were defined as submissions with a S. Dublin culture-positive foetus from a S. Dublin unvaccinated dam where results of maternal S. Dublin serology were available. Controls (n = 415) were defined as submissions where an alternative diagnosis other than S. Dublin was made in a foetus from an S. Dublin unvaccinated dam where the results of maternal S. Dublin serology were available. A logistic regression model was fitted to the data: the dichotomous dependent variable was the S. Dublin foetal culture result, and the independent variables were the maternal serum agglutination test (SAT) titre results. Salmonella serology correctly classified 87% of S. Dublin culture-positive foetuses at a predicted probability threshold of 0.44 (cut-off at which sensitivity and specificity are at a maximum, J = 0.67). The sensitivity of the SAT at the same threshold was 73.8% (95% CI: 67.4%-79.5%), and the specificity was 93.2% (95% CI: 90.3%-95.4%). The positive and negative predictive values were 84.9% (95% CI: 79.3%-88.6%) and 87.3% (95% CI: 83.5%-91.3%), respectively. This study illustrates that the use of predicted probability values, rather than the traditional arbitrary breakpoints of negative, inconclusive and positive, increases the diagnostic value of the maternal SAT. Veterinary laboratory diagnosticians and veterinary practitioners can recover from the test results, information previously categorized, particularly from those results declared to be inconclusive.

Significant re-emergence and recirculation of Schmallenberg virus in previously exposed dairy herds in Ireland in 2016.

Schmallenberg virus (SBV) circulation was investigated in 25 previously exposed dairy herds in Ireland in 2016. A population of 1,550 spring-2014-born animals, which had been monitored for SBV infection in 2014 and 2015 as part of a previous SBV surveillance study, were resampled for evidence of SBV infection during 2016. A total of 366 blood samples were collected in the 25 study herds (15 samples per herd) between 3 March 2017 and 10 March 2017 (before the 2017 vector-active season) and analysed for SBV antibodies using a competitive ELISA kit (IDVet). A total of 256 animals tested seropositive, an AP of 69.9% (95% CI: 65.1-74.4) and TP of 77.7% (95% CI: 72.3%-82.8%) when correcting for imperfect test characteristics. These results demonstrate that a new epidemic of SBV circulation occurred in these previously exposed herds in Ireland in 2016.

Schmallenberg virus: Predicting within-herd seroprevalence using bulk-tank milk antibody titres and exploring individual animal antibody titres using empirical distribution functions (EDF).

Schmallenberg virus (SBV) is transmitted by Culicoides spp. biting midges and can cause abortions and congenital malformations in ruminants and milk drop in dairy cattle. Estimating true within-herd seroprevalence is an essential component of efficient and cost-effective SBV surveillance programs. The objectives of this study were: (1) determine the correlation between bulk-tank milk (BTM)-ELISA results and within-herd seroprevalence, (2) evaluate the ability of BTM-ELISA results to predict within-herd seroprevalence and (3) explore the distributions of individual animal serology results using novel statistical methodology. BTM samples (n=24) and blood samples (n=4019) collected from all lactating cows contributing to the BTM in 26 Irish dairy herds (58-444 cows/herd) in 2014 located in a region exposed to SBV in 2012/2013, were analysed for SBV-specific antibodies using IDVet® ELISA kits. The correlation between BTM-ELISA results and within-herd seroprevalence was determined by calculating Pearson's correlation coefficient. Linear regression models were used to assess the ability of BTM-ELISA results to predict within-herd seroprevalence. The distributions of individual animal serology results were explored by determining the empirical distribution functions (EDF) of the individual animal serum ELISA results in each herd. EDFs were compared pairwise across herds, using the Kolmogorov-Smirnov statistical test. Herds with similar BTM-ELISA results, herds with similar within-herd seroprevalence and herds with similar mean-herd serology ELISA results were stratified in order to explore their respective paired-herd EDF comparisons. Statistical significance was set at p<0.05. Twenty-two herds were BTM-ELISA-positive (within-herd seroprevalence 30.6-100%) and two herds were BTM-ELISA-negative (within-herd seroprevalence 10.7 and 16.2%) indicating BTM-ELISA-negative herds can have seropositive animals present. BTM-ELISA results were highly correlated (r=0.807, p<0.0001) with, and predictive of (R2=0.832, p<0.0001) of within-herd seroprevalence. Predictions were most accurate for upper-range BTM-ELISA antibody titres, while they were less accurate at higher and lower antibody titres. This is likely a result of the overall high within-herd seroprevalence. In herds with similar BTM-ELISA results 82% of the paired-herd EDF comparisons were significantly different. In herds with similar within-herd seroprevalence and in herds with similar mean-herd serology ELISA results, 46% and 47% of the paired-herd EDF comparisons were significantly different, respectively. These results demonstrate that BTM antibody titres are highly predictive of within-herd seroprevalence in an SBV exposed region. Furthermore, exploring the serum EDFs revealed that the variation observed in the predicted within-herd seroprevalence in the regression models is likely a result of individual animal variation in serum antibody titres in these herds.

Post-epidemic Schmallenberg virus circulation: parallel bovine serological and Culicoides virological surveillance studies in Ireland.

BACKGROUND: Schmallenberg virus (SBV) emerged in northern-Europe in 2011 resulting in an epidemic of ruminant abortions and congenital malformations throughout the continent. In the years following the epidemic there have been reports of SBV overwintering and continued circulation in several European countries. When the population-level of immunity declines in exposed regions, re-introduction of SBV could result in further outbreaks of Schmallenberg disease. The aims of this study were to determine the SBV seroprevalence in previously exposed Irish dairy herds in 2014 and to investigate if SBV continued to circulate in these herds in the three years (2013-2015) following the Irish Schmallenberg epidemic. Whole-herd SBV serosurveillance was conducted in 26 herds before (spring) and following the 2014 vector-season (winter), and following the 2015 vector-season (winter). In spring 2014, 5,531 blood samples were collected from 4,070 cows and 1,461 heifers. In winter 2014, 2,483 blood samples were collected from 1,550 youngstock (8-10 months old) and a subsample (n = 933; 288 cows, 645 heifers) of the seronegative animals identified in the spring. Youngstock were resampled in winter 2015. Culicoides spp. were collected in 10 herds during the 2014 vector-season and analysed for SBV; a total of 138 pools (3,048 Culicoides) from 6 SBV vector species were tested for SBV RNA using real-time PCR. RESULTS: In spring 2014, animal-level seroprevalence was 62.5 % (cows = 84.7 %; heifers = 0.6 %). Within-herd seroprevalence ranged widely from 8.5 %-84.1 % in the 26 herds. In winter 2014, 22 animals (0.9 %; 10 cows, 5 heifers, 7 youngstock) originating in 17 herds (range 1-4 animals/herd) tested seropositive. In winter 2015 all youngstock, including the 7 seropositive animals in winter 2014, tested seronegative suggesting their initial positive result was due to persistence of maternal antibodies. All of the Culicoides pools examined tested negative for SBV-RNA. CONCLUSIONS: SBV appears to have recirculated at a very low level in these herds during 2013 and 2014, while there was no evidence of SBV infection in naïve youngstock during 2015. A large population of naïve animals was identified and may be at risk of infection in future years should SBV re-emerge and recirculate as it has done in continental Europe.