Comparison and interobserver reliability between a visual analog scale and the Wisconsin Calf Health Scoring Chart for detection of respiratory disease in dairy calves.

Respiratory disease is an ongoing challenge for calves in the dairy sector with a relatively high prevalence and impact on welfare and economics. Applying scoring protocols for detecting respiratory disease requires that they are easily implemented, consistent between observers and fast to use in daily management. This study was conducted in one Danish dairy farm from September 2020 through January 2021. The study included 126 heifer calves enrolled in the age of 17 to 24 d. All calves were observed every second day for a period of 46 d. At each visit all calves were scored with a new visual analog scale (VAS) and the Wisconsin Calf Health Scoring Chart (WCHSC). We calculated agreement between the 2 scoring systems based on conditional probability to score higher or lower than a cutoff in the VAS compared with a specified cutoff in WCHSC used as reference test. A generalized mixed effects regression model was developed to estimate the prevalence of respiratory disease and the overall agreement between the 2 scoring systems. The overall agreement between the VAS and WCHSC was 89.6%. The second part of the study assessed interobserver reliability between 2 experienced observers and between an experienced observer and veterinary students. The interobserver reliability was calculated by intraclass correlation coefficient and was 0.58 between experienced observers and was 0.34 between an experienced observer and veterinary students indicating a moderate to poor reliability between the observers. It was possible to use VAS as an alternative clinical scoring method, which primarily focuses on the general condition of the individual calf rather than specific categories of clinical signs. Our study set up lacked a comparison to other diagnostic tools i.e., thoracic ultrasound to confirm the findings which should be considered in future studies when exploring VAS as a screening tool for detection of respiratory disease in dairy calves.

Assessment of Evaluation Tools for Integrated Surveillance of Antimicrobial Use and Resistance Based on Selected Case Studies.

Regular evaluation of integrated surveillance for antimicrobial use (AMU) and resistance (AMR) in animals, humans, and the environment is needed to ensure system effectiveness, but the question is how. In this study, six different evaluation tools were assessed after being applied to AMU and AMR surveillance in eight countries: (1) ATLASS: the Assessment Tool for Laboratories and AMR Surveillance Systems developed by the Food and Agriculture Organization (FAO) of the United Nations, (2) ECoSur: Evaluation of Collaboration for Surveillance tool, (3) ISSEP: Integrated Surveillance System Evaluation Project, (4) NEOH: developed by the EU COST Action "Network for Evaluation of One Health," (5) PMP-AMR: The Progressive Management Pathway tool on AMR developed by the FAO, and (6) SURVTOOLS: developed in the FP7-EU project "RISKSUR." Each tool was scored using (i) 11 pre-defined functional aspects (e.g., workability concerning the need for data, time, and people); (ii) a strengths, weaknesses, opportunities, and threats (SWOT)-like approach of user experiences (e.g., things that I liked or that the tool covered well); and (iii) eight predefined content themes related to scope (e.g., development purpose and collaboration). PMP-AMR, ATLASS, ECoSur, and NEOH are evaluation tools that provide a scoring system to obtain semi-quantitative results, whereas ISSEP and SURVTOOLS will result in a plan for how to conduct evaluation(s). ISSEP, ECoSur, NEOH, and SURVTOOLS allow for in-depth analyses and therefore require more complex data, information, and specific training of evaluator(s). PMP-AMR, ATLASS, and ISSEP were developed specifically for AMR-related activities-only ISSEP included production of a direct measure for "integration" and "impact on decision making." NEOH and ISSEP were perceived as the best tools for evaluation of One Health (OH) aspects, and ECoSur as best for evaluation of the quality of collaboration. PMP-AMR and ATLASS seemed to be the most user-friendly tools, particularly designed for risk managers. ATLASS was the only tool focusing specifically on laboratory activities. Our experience is that adequate resources are needed to perform evaluation(s). In most cases, evaluation would require involvement of several assessors and/or stakeholders, taking from weeks to months to complete. This study can help direct future evaluators of integrated AMU and AMR surveillance toward the most adequate tool for their specific evaluation purpose.

Estimating Clinically Relevant Cut-Off Values for a High-Throughput Quantitative Real-Time PCR Detecting Bacterial Respiratory Pathogens in Cattle.

Bovine respiratory disease (BRD) results from interactions between pathogens, environmental stressors, and host factors. Obtaining a diagnosis of the causal pathogens is challenging but the use of high-throughput real-time PCR (rtPCR) may help target preventive and therapeutic interventions. The aim of this study was to improve the interpretation of rtPCR results by analysing their associations with clinical observations. The objective was to develop and illustrate a field-data driven statistical method to guide the selection of relevant quantification cycle cut-off values for pathogens associated with BRD for the high-throughput rtPCR system "Fluidigm BioMark HD" based on nasal swabs from calves. We used data from 36 herds enrolled in a Danish field study where 340 calves within pre-determined age-groups were subject to clinical examination and nasal swabs up to four times. The samples were analysed with the rtPCR system. Each of the 1,025 observation units were classified as sick with BRD or healthy, based on clinical scores. The optimal rtPCR results to predict BRD were investigated for Pasteurella multocida, Mycoplasma bovis, Histophilus somni, Mannheimia haemolytica, and Trueperella pyogenes by interpreting scatterplots and results of mixed effects logistic regression models. The clinically relevant rtPCR cut-off suggested for P. multocida and M. bovis was ≤ 21.3. For H. somni it was ≤ 17.4, while no cut-off could be determined for M. haemolytica and T. pyogenes. The demonstrated approach can provide objective support in the choice of clinically relevant cut-offs. However, for robust performance of the regression model sufficient amounts of suitable data are required.

Veterinary Herd Health Consultancy and Antimicrobial Use in Dairy Herds.

The globally increasing level of antimicrobial resistance affects both human and animal health, why it is necessary to identify ways to change our current use of antimicrobials. The veterinary herd health collaboration between veterinarians and dairy farmers provides a useful setting for changing antimicrobial use in livestock. However, farmers and veterinarians work in a complex agricultural setting influenced by socio-economic factors, which complicates their choices regarding antimicrobial usage. It is therefore necessary to be aware of the range of potential influencing factors and to integrate this knowledge in the relevant local settings. This manuscript presents a literature review of relevant factors relating to antimicrobial use within the veterinary herd health consultancy setting, including knowledge gaps of relevance for changing the use of antimicrobials. An enriched version of the framework of the Theory of Planned Behaviour was used to organise the literature review. We identified diverging attitudes on correct treatment practices and perceptions of antimicrobial resistance among veterinarians and farmers, influenced by individual risk perception as well as social norms. Furthermore, disagreements in terms of goal setting and in the frequency of herd visits in relation to herd health consultancy can negatively influence the collaboration and the intention to change antimicrobial use. Farmers and veterinarians emphasise the importance of legislation and the role of the dairy industry in changing antimicrobial use, but the relevance of specific factors depends on the country-specific context. Overall, farmers and veterinarians must communicate better to understand each other's perspectives and establish common goals within the collaboration if they are to work efficiently to reduce antimicrobial use. Farmers and veterinarians both requested changes in individual behaviour; however, they also called for national and structural solutions in terms of balanced legislation and the availability of better diagnostics to facilitate a change in antimicrobial use practices. These various paths to achieving the desired changes in antimicrobial use illustrate the need to bridge methodological research approaches of veterinary science and social sciences for a better understanding of our potential to change antimicrobial use within the dairy farm animal sector.

Epidemiology of Salmonella enterica Serovar Dublin in Cattle and Humans in Denmark, 1996 to 2016: a Retrospective Whole-Genome-Based Study.

Salmonella enterica serovar Dublin is a cattle-adapted S. enterica serovar causing both intestinal and systemic infection in its bovine host, and it is also a serious threat to human health. The present study aimed to determine the population structure of S Dublin isolates obtained from Danish cattle herds and to investigate how cattle isolates relate to Danish human isolates, as well as to non-Danish human and bovine isolates. Phylogenetic analysis of 197 Danish cattle isolates from 1996 to 2016 identified three major clades corresponding to distinct geographical regions of cattle herds. Persistence of closely related isolates within the same herd and their circulation between epidemiologically linked herds for a period of more than 20 years were demonstrated. These findings suggest that a lack of internal biosecurity and, to some extent, also a lack of external biosecurity in the herds have played an important role in the long-term persistence of S Dublin in Danish cattle herds in the period investigated. Global population analysis revealed that Danish cattle isolates clustered separately from bovine isolates from other countries, whereas human isolates were geographically spread. Resistance genes were not commonly demonstrated in Danish bovine isolates; only the isolates within one Danish clade were found to often harbor two plasmids of IncFII/IncFIB and IncN types, the latter plasmid carrying blaTEM-1, tetA, strA, and strB antibiotic resistance genes.IMPORTANCES Dublin causes economic losses in cattle production, and the bacterium is a public health concern. A surveillance and control program has been in place in Denmark since 2002 with the ultimate goal to eradicate S Dublin from Danish cattle herds; however, a small proportion of herds have remained positive for many years. In this study, we demonstrate that herds with persistent infection often were infected with the same strain for many years, indicating that internal biosecurity has to be improved to curb the infection. Further, domestic cases of S Dublin infection in humans were found to be caused both by Danish cattle isolates and by isolates acquired abroad. This study shows the strength of whole-genome sequencing to obtain detailed information on epidemiology of S Dublin and allows us to suggest internal biosecurity as a main way to control this bacterium in Danish cattle herds.

A One Health Evaluation of the University of Copenhagen Research Centre for Control of Antibiotic Resistance.

We applied the evaluation framework developed by the EU COST Action "Network of Evaluation of One Health" (NEOH) to assess the operations, supporting infrastructures and outcomes of a research consortium "University of Copenhagen Research Centre for Control of Antibiotic Resistance" (UC-CARE). This 4-year research project was a One Health (OH) initiative with participants from 14 departments over four faculties as well as stakeholders from industry and health authorities aiming to produce new knowledge to reduce the development of antimicrobial resistance (AMR). This was a case study focusing on assessing beneficial and counter-productive characteristics that could affect the OH outcomes. The study was also used to provide feedback to NEOH about the evaluation framework. The framework and evaluation tools are described in the introduction paper of this special journal issue. Data for the evaluation were extracted from the funding research proposal, the mid-term UC-CARE project evaluation report and supplemented with opinions elicited from project participants and stakeholders. Here, we describe the underlying system, theory of change behind the initiative and adapted questions from the NEOH tools that we used for semi-open interviews with consortium members throughout the evaluation process. An online survey was used to obtain information from stakeholders. The NEOH evaluation tools were then used for the qualitative and quantitative evaluation of the OH characteristics of UC-CARE. Senior UC-CARE researchers were interested and willing to be interviewed. Young scientists were more difficult to engage in interviews, and only 25% of stakeholders answered the online survey. Interviewees mentioned that the main benefit of UC-CARE was an increased awareness and general understanding of AMR issues. All interviewees stated that the adopted OH approach was relevant given the complexity of AMR. However, some questioned the applicability, and identified potentially counter-productive issues mainly related to the information sharing, collaboration and working methods across the consortium. A more integrated project organization, more stakeholder involvement and time for the project, flexibility in planning and a dedicated OH coordinator were suggested to allow for more knowledge exchange, potentially leading to a higher societal impact.

Application of the NEOH Framework for Self-Evaluation of One Health Elements of a Case-Study on Obesity in European Dogs and Dog-Owners.

Obesity is a malnutrition disorder of global concern with increasing prevalence driven by underlying societal, economic and environmental mechanisms leading to changed physical activity patterns, eating behaviors and diet compositions in both humans and in their pet-dogs. A questionnaire-based study was carried out as a joint effort across 11 European countries. It was considered a One Health (OH) initiative between scientists from human and animal health sectors aiming to identify factors associated with obesity in dog owners and their dogs. Expected outcomes of this approach included new insights unachievable by single-sector research initiatives, and hence potentially leading to new cross-sectorial solutions. We performed an internal evaluation among the actors of the obesity initiative using the framework for evaluation developed by the "Network for Evaluation of One Health" (NEOH). It served as a case-study for the NEOH consortium to illustrate the application and provide feedback on the utility of the framework. The evaluation was performed by a subgroup of scientists also involved in the obesity study group, and it consisted of: (1) the definition of the initiative and its context, (2) the description of the theory of change, and (3) the qualitative and quantitative process evaluation of operations and supporting infrastructures scored on a scale from 0 to 1. In the One Health operations, the obesity study initiative scored medium high on OH-thinking (0.5) and OH-planning (0.45), and relatively high on OH-working (0.7). The supporting infrastructure score was high for systemic organization (0.8), but low for sharing (0.45) and learning (0.28). The calculated OH-index was 0.29 (on scale 0 to 1) indicating that the full potential of health integration and collaboration was not exploited in the initiative, and the main issue identified was a lack of stakeholder engagement. The OH-ratio of 1.1 indicated equal focus on operations and supporting infrastructures. Hence, the evaluation identified potentially counterproductive as well as beneficial characteristics, which are further discussed in this paper in relation to the expected outcomes. The NEOH framework for evaluation requires that the evaluators have a good understanding of systems thinking and the mechanisms of the health issue targeted by the initiative.

A longitudinal observational study of the dynamics of Mycoplasma bovis antibodies in naturally exposed and diseased dairy cows.

Mycoplasma bovis is an important pathogen causing disease and substantial economic losses in cattle. However, knowledge of the dynamics of antibody responses in individual cows in the face of an outbreak is currently extremely limited. The use of commercial antibody tests to support clinical decision-making and for surveillance purposes is therefore challenging. Our objective was to describe the dynamics of M. bovis antibody responses in 4 Danish dairy herds experiencing an acute outbreak of M. bovis-associated disease, and to compare the antibody dynamics between dairy cows with different disease manifestations. A total of 120 cows were examined using a standardized clinical protocol and categorized into 4 disease groups: "mastitis," "systemic," "nonspecific," and "none." Paired blood and milk samples were collected and tested using a commercial M. bovis antibody-detecting ELISA. Plots of raw data and generalized additive mixed models with cow and herd as random effects were used to describe serum and milk antibody dynamics relative to the estimated time of onset of clinical disease. Cows with mastitis had high optical density measurement (ODC%) of antibodies in both milk and serum at disease onset. The estimated mean ODC% in milk was below the manufacturer's cut-off for the other groups for the entire study period. The estimated mean serum ODC% in the "systemic" group was high at onset of disease and stayed above the cut-off until 65 d after disease onset. However, the lower 95% confidence interval (CI) for the mean ODC% was only above the manufacturer's cut-off between 7 and 17 d after onset of disease. The CI of the "systemic" and "none" groups did not overlap at any time between the day of disease onset and 65 d after disease onset, and the estimated mean ODC% for both the "nonspecific" and "none" groups were generally below the cut-off for the majority of the study period. In conclusion, the serum antibody responses were highly dynamic and showed a high level of variation between individual cows. This strongly suggests that serology is unlikely to be useful for individual diagnosis of M. bovis-associated disease in dairy cows. However, it might still be useful for herd- or group-level diagnosis. Antibodies in milk were only increased in cows with M. bovis mastitis, indicating that milk antibody measurements only have diagnostic utility for cows with mastitis.

Veterinary Expert Opinion on Potential Drivers and Opportunities for Changing Antimicrobial Usage Practices in Livestock in Denmark, Portugal, and Switzerland.

Reducing antimicrobial use (AMU) in livestock is requested by Public Health authorities. Ideally, this should be achieved without jeopardizing production output or animal health and welfare. Thus, efficient measures must be identified and developed to target drivers of AMU. Veterinarians play a central role in the identification and implementation of such efficient interventions. Sixty-seven veterinarians with expertise in livestock production in Denmark, Portugal, and Switzerland participated in an expert opinion study aimed at investigating experiences and opinions of veterinarians about the driving forces and practices related to AMU in the main livestock sectors (broiler, dairy cattle, fattening/veal calf, and pig industry) of the aforementioned countries. Opinions on potential factors influencing the choice of antimicrobials and opportunities to reduce AMU were collected. Antibiograms are seldom used, mainly due to the time lag between testing and obtaining the results. The perceived percentage of treatment failures varied between countries and livestock sectors; however, little changes were reported over time (2005-2015). The animal health problems of each livestock sector most frequently leading to AMU did not vary substantially between countries. Mandatory official interventions (i.e., binding measures applied by national or international authorities) were highlighted as having the biggest impact on AMU. There was a variation in the experts' opinion regarding feasibility and impact of interventions both between countries and livestock sectors. Nevertheless, improved biosecurity and education of veterinarians frequently received high scores. Most veterinarians believed that AMU can be reduced. The median potential reduction estimates varied from 1% in Swiss broilers to 50% in Portuguese broilers and veal/fattening calves in all countries. We hypothesize that the differences in views could be related to disease epidemiology, animal husbandry, and socio-economic factors. A profound investigation of these disparities would provide the required knowledge for developing targeted strategies to tackle AMU and consequently resistance development. However, experts also agreed that mandatory official interventions could have the greatest impact on antimicrobial consumption. Furthermore, improvement of biosecurity and education of veterinarians, the use of zinc oxide (in pigs), improving vaccination strategies, and the creation of treatment plans were the measures considered to have the largest potential to reduce AMU. This paper can inform policymakers in Europe and countries with a similar animal production regarding their AMU policy.

Comparison of Antimicrobial Consumption Patterns in the Swiss and Danish Cattle and Swine Production (2007-2013).

Veterinary antimicrobial consumption patterns vary considerably across Europe. These differences are not only limited to the total amount consumed but are also observed with regards to the relative proportion of the various antimicrobial classes used. Currently, most of the data on veterinary antimicrobials are reported at sales level without any information on the consumption by different animal species. This hinders a proper comparison of antimicrobial consumption at the species level between countries. However, it is imperative to improve our understanding on antimicrobial usage patterns at the species level, as well as on the drivers contributing to those differences. This will allow for development of tailored interventions with the lowest possible risk for human health, while ensuring effective treatment of diseased livestock. An important step to attain such an objective is to perform detailed comparisons of the antimicrobial consumption in each species between countries. We compared antimicrobial consumption estimates for cattle and pigs in Switzerland and Denmark, in order to distinguish species-specific patterns and trends in consumption from 2007 to 2013. Swiss data were obtained from a previous study that assessed methodologies to stratify antimicrobial sales per species; Danish antimicrobial consumption estimates were assembled from Danish Integrated Antimicrobial Resistance Monitoring and Research Programme reports. A decrease in antimicrobial consumption in milligrams per kilogram of biomass was observed for both countries (4.5% in Denmark and 34.7% in Switzerland) when comparing 2013 to 2007. For pigs and cattle, the overall consumption per kilogram of biomass of most antimicrobial classes was higher in Switzerland than in Denmark. Large variations in the relative consumption of different antimicrobial classes were also evident. Sulfonamides/trimethoprim and tetracyclines were consumed in a higher proportion in Switzerland than in Denmark, whereas the relative consumption of penicillins was higher in Denmark. The differences observed in veterinary antimicrobial consumption are not solely related to animal demographic characteristics in these two countries. Other factors, such as the level of biosecurity and farming practices, veterinarians and farmers' education, or governmental/industry programs put in place might also partly explain these variations. These differences should be taken into account when aiming to implement targeted interventions to reduce antimicrobial consumption.

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.

Exposure assessment of extended-spectrum beta-lactamases/AmpC beta-lactamases-producing Escherichia coli in meat in Denmark.

INTRODUCTION: Extended-spectrum beta-lactamases (ESBL) and AmpC beta-lactamases (AmpC) are of concern for veterinary and public health because of their ability to cause treatment failure due to antimicrobial resistance in Enterobacteriaceae. The main objective was to assess the relative contribution (RC) of different types of meat to the exposure of consumers to ESBL/AmpC and their potential importance for human infections in Denmark. MATERIAL AND METHODS: The prevalence of each genotype of ESBL/AmpC-producing E. coli in imported and nationally produced broiler meat, pork and beef was weighted by the meat consumption patterns. Data originated from the Danish surveillance program for antibiotic use and antibiotic resistance (DANMAP) from 2009 to 2011. DANMAP also provided data about human ESBL/AmpC cases in 2011, which were used to assess a possible genotype overlap. Uncertainty about the occurrence of ESBL/AmpC-producing E. coli in meat was assessed by inspecting beta distributions given the available data of the genotypes in each type of meat. RESULTS AND DISCUSSION: Broiler meat represented the largest part (83.8%) of the estimated ESBL/AmpC-contaminated pool of meat compared to pork (12.5%) and beef (3.7%). CMY-2 was the genotype with the highest RC to human exposure (58.3%). However, this genotype is rarely found in human infections in Denmark. CONCLUSION: The overlap between ESBL/AmpC genotypes in meat and human E. coli infections was limited. This suggests that meat might constitute a less important source of ESBL/AmpC exposure to humans in Denmark than previously thought - maybe because the use of cephalosporins is restricted in cattle and banned in poultry and pigs. Nonetheless, more detailed surveillance data are required to determine the contribution of meat compared to other sources, such as travelling, pets, water resources, community and hospitals in the pursuit of a full source attribution model.

Effects of experimental immunosuppression in cattle with persistently high antibody levels to Salmonella Dublin lipopolysaccharide O-antigens.

BACKGROUND: Salmonella Dublin (S. Dublin) is a zoonotic bacterium which is host adapted to cattle. The bacterium can cause subclinical persistent infection in cattle (carriers), which may be reactivated. During reactivation, animals may shed bacteria, thus constituting a source of infection for other animals. Identification of such carriers is assumed to be critical in attempts to control and eradicate the infection. Some authors suggest that persistently high antibody levels in serum or milk is indicative of a carrier state in cattle. However, this has been questioned by other studies in which S. Dublin were not found in all animals suspected of being carriers based on antibody measurements when such animals were examined at slaughter. Some hypothesize that the lack of isolated bacteria from long-term high antibody level cattle is due to a latent infection stage that can later be reactivated, for instance during stress around calving or due to transportation. This study examined nine adult cattle with persistently high antibody responses to S. Dublin O-antigen based lipopolysaccharide for cultivable bacteria in faeces, milk and internal organs before and after transportation, isolation and experimental immunosuppression with dexamethasone sodium phosphate over a period of 7-14 days. RESULTS: Clear signs of immunosuppression were seen as expression of leucocytosis and neutrophilia in all animals on day 3-5 after the first injections with dexamethasone sodium phosphate. No clinical signs or necropsy findings indicating salmonellosis were observed in any of the animals. No shedding of S. Dublin was found in faeces (collected four times daily) or milk (collected twice daily) at any point in time during the 7-14 day period. S. Dublin was recovered by a conventional culture method from tissue samples from mammary lymph nodes, spleen and liver collected from three animals at necropsy. CONCLUSION: In this study, immunosuppression by transportation stress or dexamethasone treatment did not lead to excretion of S. Dublin in milk or faeces from infected animals. The study questions the general conception that cattle with persistently high antibody levels against S. Dublin O-antigens in naturally infected herds should be considered high risk for transmission and therefore culled as part of effective intervention strategies. It is suggested that the location of S. Dublin infected foci in the animal plays a major role for the risk of excreting bacteria.