Using registry data to identify individual dairy cows with abnormal patterns in routinely recorded somatic cell counts.

Data from the Danish milk recording system routinely enter the Danish Cattle Database, including somatic cell counts (SCC) for individual animals. Elevated SCC can signal intramammary inflammation, suggesting subclinical mastitis. Detecting mastitis is pivotal to limit severity, prevent pathogen spread, and target treatment or culling. This study aimed to differentiate normal and abnormal SCC patterns using recorded registry data. We used registry data from 2010 to 2020 for dairy cows in herds with 11 annual milk recordings. To create consistency across herds, we used data from 13,996 unique animals and eight different herds, selected based on the amount of data available, only selecting Holstein animals and conventional herds. We fitted log10-transformed SCC to days in milk (DIM) using the Wilmink and Wood's curve functions, originally developed for milk yield over the lactation. We used Nonlinear Least Square and Nonlinear Mixed Effect models to fit the log10-transformed SCC observations to DIM at animal level. Using mean squared residuals (MSR), we found a consistently better fit using a Wood's style function. Detection of MSR outliers in the model fitting process was used to identify animals with log10(SCC) curves deviating from the expected "normal" curve for that same animal. With this study, we propose a method to identify single animals with SCC patterns that indicate abnormalities, such as mastitis, based on registry data. This method could potentially lead to a registry data-based detection of mastitis cases in larger dairy herds.

Use of Danish National Somatic Cell Count Data to Assess the Need for Dry-Off Treatment in Holstein Dairy Cattle.

In Denmark, PCR testing of dairy cattle is commonly used to select animals for the antibacterial treatment of intramammary infection (IMI) during the dry-off period. IMI is associated with a high somatic cell count (SCC), routinely recorded for milk quality control for most commercial dairy herds. This study aimed to compare SCC curves over the lactation among dairy cows with positive vs. negative PCR test results for four major IMI pathogens. Data from 133,877 PCR-tested Holstein cows from 1364 Danish conventional dairy herds were used to fit a nonlinear mixed-effects model using a modified four-parameter Wilmink function. We stratified the data into first, second, third or fourth and later parity and fitted Wilmink curves to all SCC observations between 6 and 305 days in milk. The PCR tests were taken before dry-off at the end of the lactation to investigate which animals qualified for selective dry cow therapy. A PCR Ct-value of 37 and below was used to determine if an animal was PCR positive for any of the following IMI pathogens: Staphylococcus aureus, Streptococcus agalactiae, Str. dysgalactiae and Str. uberis. Our findings showed that mean SCC curve fits were higher for PCR-positive animals in all four parity groups and across lactations. The use of SCC data fitted to the entire lactation for multiple lactations enabled quantification of overall differences in SCC curves between cattle with and without detected IMI, adjusted for parity group and stage of lactation. These findings are relevant to the use of SCC to support treatment decisions.

Estimating true prevalence through questionnaire data.

We present a general analytical method for obtaining unbiased prevalence estimates based on data from regional or national testing programs, where individual participation in the testing program is voluntary but where additional questionnaire data is collected regarding the individual-level reason/motivation for being tested. The approach is based on re-writing the conditional probabilities for being tested, being infected, and having symptoms, so that a series of equations can be defined that relate estimable quantities (from test data and questionnaire data) to the result of interest (an unbiased estimate of prevalence). The final estimates appear to be robust based on prima-facie examination of the temporal dynamics estimated, as well as agreement with an independent estimate of prevalence. Our approach demonstrates the potential strength of incorporating questionnaires when testing a population during an outbreak, and can be used to help obtain unbiased estimates of prevalence in similar settings.

Effect of vaccination on household transmission of SARS-CoV-2 Delta variant of concern.

Effective vaccines protect individuals by not only reducing the susceptibility to infection, but also reducing the infectiousness of breakthrough infections in vaccinated cases. To disentangle the vaccine effectiveness against susceptibility to infection (VES) and vaccine effectiveness against infectiousness (VEI), we took advantage of Danish national data comprising 24,693 households with a primary case of SARS-CoV-2 infection (Delta Variant of Concern, 2021) including 53,584 household contacts. In this setting, we estimated VES as 61% (95%-CI: 59-63), when the primary case was unvaccinated, and VEI as 31% (95%-CI: 26-36), when the household contact was unvaccinated. Furthermore, unvaccinated secondary cases with an infection exhibited a three-fold higher viral load compared to fully vaccinated secondary cases with a breakthrough infection. Our results demonstrate that vaccinations reduce susceptibility to infection as well as infectiousness, which should be considered by policy makers when seeking to understand the public health impact of vaccination against transmission of SARS-CoV-2.

Electronic reporting of diagnostic laboratory test results from all healthcare sectors is a cornerstone of national preparedness and control of COVID-19 in Denmark.

The COVID-19 pandemic has led to an unprecedented demand for real-time surveillance data in order to inform critical decision makers regarding the management of the pandemic. The aim of this review was to describe how the Danish national microbiology database, MiBa, served as a cornerstone for providing data to the real-time surveillance system by linkage to other nationwide health registries. The surveillance system was established on an existing IT health infrastructure and a close network between clinical microbiologists, information technology experts, and public health officials. In 2020, testing capacity for SARS-CoV-2 was ramped up from none to over 10,000 weekly PCR tests per 100,000 population. The crude incidence data mirrored this increase in testing. Real-time access to denominator data and patient registries enabled adjustments for fluctuations testing activity, providing robust data on crude SARS-CoV-2 incidence during the changing diagnostic and management strategies. The use of the same data for different purposes, for example, final laboratory reports, information to the public, contact tracing, public health, and science, has been a critical asset for the pandemic response. It has also raised issues concerning data protection and critical capacity of the underlying technical systems and key resources. However, even with these limitations, the setup has enabled decision makers to adopt timely interventions. The experiences from COVID-19 may motivate a transformation from traditional indicator-based public health surveillance to an all-encompassing information system based on access to a comprehensive set of data sources, including diagnostic and reference microbiology.

Livestock-associated MRSA survival on house flies (Musca domestica) and stable flies (Stomoxys calcitrans) after removal from a Danish pig farm.

We caught stable- and house flies on a Danish LA-MRSA positive pig farm. Stable- and house flies were housed together and culled over time to test for the presence of live LA-MRSA bacteria at 24 h intervals to establish the length of time for which LA-MRSA can persist on flies. On average, 7% of stable flies and 27% of house flies tested positive for LA-MRSA immediately upon removal from the farm. LA-MRSA prevalence decreased over time and estimates based on a Kaplan-Meier time-to-event analysis indicated that the probability of a stable- or house fly testing positive for LA-MRSA was 5.4% and 7.8% after 24 h, 3.5% and 4.3% after 48 h, 3.1% and 2.2% after 72 h and 0.4% and 0% after 96 h of removal from the pig farm, respectively. Simultaneously, we found that caged cultivated house flies became carriers of LA-MRSA, without direct contact with pigs, in the same proportions as wild flies inside the farm. We provide distance distributions of Danish pig farms and residential addresses as well as the calculated maximum dispersal potentials of stable- and house flies, which suggest that there is a potential for stable- and house flies dispersing live LA-MRSA bacteria into the surrounding environment of a pig farm. This potential should therefore be considered when modelling the spread between farms or the risk posed to humans living in close proximity to LA-MRSA pig farm sources.

Dynamics of somatic cell count (SCC) and differential SCC during and following intramammary infections.

Somatic cell count is frequently used as an indicator of intramammary infections (IMI) in dairy cattle worldwide. The newly introduced differential SCC (DSCC) can potentially contribute to detection of IMI. The purpose of this study was to investigate the dynamics of SCC and DSCC after IMI. We used a data set with monthly samples from 2 Danish dairy herds through 1 yr, using bacterial culture to identify IMI. The dynamics of SCC and DSCC with regard to IMI were assessed at quarter level following new IMI with each of 3 defined pathogen groups, major, minor, or "other" pathogens, using general additive models. Both SCC and DSCC increased after IMI, with a more pronounced increase if major or other pathogens were detected compared with minor pathogens. We found that DSCC increased after IMI with other pathogens in both herds and, in herd 2, after IMI caused by major and minor pathogens. We also estimated the duration of increased SCC and DSCC when they exceeded a threshold, done separately for each pathogen group. Major pathogens had the longest-lasting effect in both herds for both SCC and DSCC. We conclude that the magnitude and duration of response of SCC and DSCC to IMI differs between herds and causative pathogens.

Seroprevalence of Toxoplasma gondii infection in sows and finishers from conventional and organic herds in Denmark: Implications for potential future serological surveillance.

Pigs are one of several host species for Toxoplasma gondii parasites, and consumption of infected pork may lead to toxoplasmosis in humans. We estimated seroprevalence in sows and finishers from conventional and organic herds in Denmark and discussed the strategies for reducing the risk from pork. We collected 447 blood samples from 59 herds, and additional meat-juice samples from 212 of the same pigs. Using a T. gondii IgG commercial ELISA test, we found 2% (95% CI = 0.4%-5%) apparent seroprevalence of T. gondii in conventional finishers, 11% (95% CI = 6%-17%) in organic finishers, 19% (95% CI = 11%-30%) in conventional sows and 60% (95% CI = 47%-72%) in organic sows. The odds of an animal testing positive for T. gondii was 16 times higher (95% CI = 4.6-74.3) in organic compared to conventional herds. The odds were 22 times higher (95% CI = 6.5-88.3) if the animal was a sow compared to a finisher. Meat-juice ELISA values were significantly correlated with plasma results (P < 0.001), but on average 64% of the blood-plasma ELISA values. Lowering the recommended cut-off from 20 to 13 percent positive values of the positive control for meat-juice ELISA, resulted in the meat-juice ELISA identifying 93% of the plasma positives as positive and 99% of the plasma negatives as negative. The time taken to detect one or more infected pigs from a T. gondii positive herd at slaughter was estimated using abattoir data on pigs (17,195,996) and batches (165,569) delivered to Danish abattoirs in 2018. The time to detection was affected by the seroprevalence, frequency at which the pigs were delivered, the number of samples tested per batch delivery and the batch sizes. Time to detection was long in conventional finisher herds due to low prevalence, and in sow herds because of intermittent delivery of a low number of sows. In organic finisher herds, time to detection was short due to medium prevalence and frequent delivery of a high number of finishers. Conventional finisher herds may be classified as low-risk, organic finisher herds as medium-risk, and conventional and organic sow herds as high-risk herds. Risk-mitigation strategies at processing plants (freezing or curing) or at the consumer level (heat treatment) for meat originating from high-risk herds, surveillance of medium-risk herds, and auditing for controlled housing (high biosecurity) in low-risk herds may be cost-effective alternatives to serological surveillance of all Danish pig herds.

Practical and effective diagnosis of animal anthrax in endemic low-resource settings.

Anthrax threatens human and animal health, and people's livelihoods in many rural communities in Africa and Asia. In these areas, anthrax surveillance is challenged by a lack of tools for on-site detection. Furthermore, cultural practices and infrastructure may affect sample availability and quality. Practical yet accurate diagnostic solutions are greatly needed to quantify anthrax impacts. We validated microscopic and molecular methods for the detection of Bacillus anthracis in field-collected blood smears and identified alternative samples suitable for anthrax confirmation in the absence of blood smears. We investigated livestock mortalities suspected to be caused by anthrax in northern Tanzania. Field-prepared blood smears (n = 152) were tested by microscopy using four staining techniques as well as polymerase chain reaction (PCR) followed by Bayesian latent class analysis. Median sensitivity (91%, CI 95% [84-96%]) and specificity (99%, CI 95% [96-100%]) of microscopy using azure B were comparable to those of the recommended standard, polychrome methylene blue, PMB (92%, CI 95% [84-97%] and 98%, CI 95% [95-100%], respectively), but azure B is more available and convenient. Other commonly-used stains performed poorly. Blood smears could be obtained for <50% of suspected anthrax cases due to local customs and conditions. However, PCR on DNA extracts from skin, which was almost always available, had high sensitivity and specificity (95%, CI 95% [90-98%] and 95%, CI 95% [87-99%], respectively), even after extended storage at ambient temperature. Azure B microscopy represents an accurate diagnostic test for animal anthrax that can be performed with basic laboratory infrastructure and in the field. When blood smears are unavailable, PCR using skin tissues provides a valuable alternative for confirmation. Our findings lead to a practical diagnostic approach for anthrax in low-resource settings that can support surveillance and control efforts for anthrax-endemic countries globally.

Effects of intra-articular administration of lidocaine, mepivacaine, and the preservative methyl parahydroxybenzoate on synovial fluid biomarkers of horses.

OBJECTIVE: To compare the extent of inflammation and catabolic collagen response in the middle carpal joints (MCJs) of healthy horses following intra-articular injection of 2% lidocaine, 2% mepivacaine, lactated Ringer solution (LRS), or 0.1% methyl parahydroxybenzoate. ANIMALS: 17 adult horses. PROCEDURES: In the first of 2 experiments, the left middle carpal joint (MCJ) of each of 12 horses was injected with 10 mL of 2% lidocaine (n = 3), 2% mepivacaine (3), or LRS (control; 6). After a 4-week washout period, the right MCJ of the horses that received lidocaine or mepivacaine was injected with 10 mL of LRS, and the right MCJ of horses that received LRS was injected with 10 mL of 2% lidocaine (n = 3) or 2% mepivacaine (3). In experiment 2, the left MCJ of each of 5 horses was injected with 10 mL of 0.1% methyl parahydroxybenzoate. After a 48-hour washout period, the right MCJ of each horse was injected with 10 mL of LRS. Synovial fluid (SF) samples were aseptically collected before and at predetermined times after each injection. Synovial fluid WBC count, neutrophil percentage, and total protein, neutrophil myeloperoxidase, neutrophil elastase, and Coll2-1 concentrations were compared among treatments. RESULTS: Both lidocaine and mepivacaine induced SF changes indicative of inflammation and a catabolic collagen response, but the magnitude of those changes was more pronounced for lidocaine. Methyl parahydroxybenzoate did not cause any SF changes indicative of inflammation. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that mepivacaine was safer than lidocaine for intra-articular injection in horses.

Contrasting effects of high-starch and high-sugar diets on ruminal function in cattle.

The experiment reported in this research paper aimed to determine whether clinical and subclinical effects on cattle were similar if provided with isoenergetic and isonitrogenous challenge diets in which carbohydrate sources were predominantly starch or sugar. The study was a 3 × 3 Latin square using six adult Jersey cows with rumen cannulae, over 9 weeks. In the first 2 weeks of each 3 week experimental period cows were fed with a maintenance diet and, in the last week, each animal was assigned to one of three diets: a control diet (CON), being a continuation of the maintenance diet; a high starch (HSt) or a high sugar (HSu) diet. Reticuloruminal pH and motility were recorded throughout the study period. Blood and ruminal samples were taken on day-1 (TP-1), day-2 (TP-2) and day-7 (TP-7) of each challenge week. Four clinical variables were recorded daily: diarrhoea, inappetence, depression and ruminal tympany. The effects of treatment, hour of day and day after treatment on clinical parameters were analysed using linear mixed effects (LME) models. Although both challenge diets resulted in a decline in pH, an increase in the absolute pH residuals and an increase in the number of minutes per day under pH 5.8, systemic inflammation was only detected with the HSt diet. The challenge diets differentially modified amplitude and period of reticuloruminal contractions compared with CON diet and both were associated with an increased probability of diarrhoea. The HSu diet reduced the probability of an animal consuming its complete allocation. Because the challenge diets were derived from complex natural materials (barley and molasses respectively), it is not possible to assign all the differential effects to the difference in starch and sugar concentration: non-starch components of barley or non-sugar components of molasses might have contributed to some of the observations. In conclusion, substituting much of the starch with sugar caused no substantial reduction in the acidosis load, but inflammatory response was reduced while feed rejection was increased.

Associations between udder health and culling in dairy cows.

Culling is an important management tool in dairy herds, as it affects herd economics and animal welfare. In relation to health, culling is usually studied as a consequence of health disorders, but it can also be regarded as a tool to manage health in the herd by making strategic culling decisions. In this study, we used data from the Danish Cattle Database in herd-wise survival analyses to investigate factors associated with culling, in relation to udder health, in 1,452 dairy herds. The data included milk yield, somatic cell counts (SCC), parity, and different disease related factors with a special focus on udder health. In each herd, observations and survival analyses were divided into five groups: mid lactation heifers, late lactation heifers, early lactation cows, mid lactation cows, and late lactation cows. The results showed that a high average milk yield reduced the culling hazard, and a number of risk factors (e.g., parity, a high SCC or treatment of mastitis) were associated with an increased hazard for culling. Importantly, the strength and direction of many of these associations was dependent on the lactation stage. The resulting coefficients were further analysed by principal component analysis and clustering to explore variations in culling risk factors amongst herds. In some herds, parity was an important factor for culling, while in other herds, average milk yield, SCC, or udder health were more important factors. However, clusters were substantially overlapping, indicating that the decision making process underlying culling is complex and multifactorial.

Should the Contribution of One Additional Lame Cow Depend on How Many Other Cows on the Farm Are Lame?

Welfare Quality® proposes a system for aggregation according to which the total welfare score for a group of animals is a non-linear effect of the prevalence of welfare scores across the individuals within the group. Three assumptions serve to justify this: (1) experts do not follow a linear reasoning when they assess a welfare problem; (2) it serves to prevent compensation (severe welfare problems hidden by scoring well on other aspects of welfare); (3) experts agree on the weight of different welfare measures. We use two sources of data to examine these assumptions: animal welfare data from 44 Danish dairy farms with Danish Holstein Friesian cows, and data from a questionnaire study with a convenience sample of 307 experts in animal welfare, of which we received responses from over 50%. Our main results were: (1) the total group-level welfare score as assigned by experts is a non-linear function of the individual animal welfare states within the group; (2) the WQ system does not prevent what experts perceive as unacceptable compensation; (3) the level of agreement among experts appears to vary across measures. Our findings give rise to concerns about the proposed aggregation system offered by WQ.

Associations between Antibacterial Treatment and the Prevalence of Tail-Biting-Related Sequelae in Danish Finishers at Slaughter.

Secondary infections as a result of tail biting cause substantial economic losses in pig production and are a subject of concern for animal welfare. The use of first-choice antibacterial agents in the treatment of tail biting in finishing pigs is hypothesized to be negatively correlated with the development of systemic infection. This would be expected to reduce the prevalence of post-mortem pyemic sequelae (such as osteomyelitis and abscesses) in finishers with tail-bite lesions. We performed a register-based study that included three Danish databases, holding information on the purchase of antibacterials at herd level (VetStat), herd demographics (Central Husbandry Register), and relevant observations at slaughter (meat inspection data). We included all finishers from indoor production finisher herds that met the inclusion criterion of at least one slaughtered finisher with a recorded tail-bite observation during 2015 at the single largest Danish abattoir. The final dataset held 1,070 herds with one or more tail-bite observations, from which 14,411 of 2,906,626 finishers (0.50%) had an individual record of a tail bite. Within this group of finishers with tail-bite observations, the recorded tail-biting-related sequelae included osteomyelitis (8.1%), abscesses in the hindquarters (10.5%), abscesses in the forequarters (2.3%), abscesses in the mid-section of the carcass (2.9%), abscesses in the limbs (2.4%), and chronic arthritis (0.5%). Due to a high-herd prevalence (>25%), osteomyelitis and abscesses in the hindquarters were selected for further analysis. The occurrence of osteomyelitis and hindquarter abscesses in individual finishers with tail-bite observations was described using a generalized linear mixed effects model with binomial response and logit link. Herd was included as a random effect, while herd size and various antibacterial treatments were tested for inclusion in the model as fixed effects. The final models indicated a significant association between herd size and both osteomyelitis (p = 0.014) and hindquarter abscesses (p < 0.001), with larger herds (2,001-12,000 registered finisher pigs) showing a reduced risk. Further, a negative association was found between the occurrence of hindquarter abscesses and the use of oral pleuromutilin (p = 0.022). The significant association with herd size highlights the potential importance of management factors in reducing the occurrence of tail-bite lesions in finishing pigs.

An Invasive Mammal (the Gray Squirrel, Sciurus carolinensis) Commonly Hosts Diverse and Atypical Genotypes of the Zoonotic Pathogen Borrelia burgdorferi Sensu Lato.

Invasive vertebrate species can act as hosts for endemic pathogens and may alter pathogen community composition and dynamics. For the zoonotic pathogen Borrelia burgdorferi sensu lato, the agent of Lyme borreliosis, recent work shows invasive rodent species can be of high epidemiological importance and may support host-specific strains. This study examined the role of gray squirrels (Sciurus carolinensis) (n = 679), an invasive species in the United Kingdom, as B. burgdorferi sensu lato hosts. We found that gray squirrels were frequently infested with Ixodes ricinus, the main vector of B. burgdorferi sensu lato in the United Kingdom, and 11.9% were infected with B. burgdorferi sensu lato. All four genospecies that occur in the United Kingdom were detected in gray squirrels, and unexpectedly, the bird-associated genospecies Borrelia garinii was most common. The second most frequent infection was with Borrelia afzelii. Genotyping of B. garinii and B. afzelii produced no evidence for strains associated with gray squirrels. Generalized linear mixed models (GLMM) identified tick infestation and date of capture as significant factors associated with B. burgdorferi sensu lato infection in gray squirrels, with infection elevated in early summer in squirrels infested with ticks. Invasive gray squirrels appear to become infected with locally circulating strains of B. burgdorferi sensu lato, and further studies are required to determine their role in community disease dynamics. Our findings highlight the fact that the role of introduced host species in B. burgdorferi sensu lato epidemiology can be highly variable and thus difficult to predict.