Progress 05/01/23 to 04/30/24

Outputs
Target Audience:This project aims to develop meaningful and actionable information among dairy-sourced calf production systems. This includes dairy-beef, veal, and dairy heifer replacement growers. The target audience for the research Aims (Aim 1 and Aim 2) include other veterinarians and researchers that have mutual research interests in antimicrobial resistance, pathogen dissemination pathways, and animal movement networks. Additionally, the target audience for Aim 3 (extension and education) includes the producers and association professionals involved with management practices and decisions within the production systems, including individuals working at collection points for dairy sourced calves (auctions and calf dealers). Changes/Problems:The first Specific Aim is focused on understanding the transmission patterns and prevention practices forSalmonellaDublin. As with any project, there have been forseen and unforseen challenges with implementation. Firstly, it is sometimes difficult toconvince producers to participate in the research project. For a project focused on Salmonella Dublin, this is somewhat easier, since producers experience direct economic consequences and recognize the value and need for research. In this case, however, the challenge is to identify a sufficient number of producers for participation. The farms are geographically broadly distributed. Unlike dairy farms which are listed on the state list of Grade A Certified farms, there is no centralized list of farms by which to contact producers. To overcome this obstacle, we've worked directly with veterinary clinics with established relationships. These veterinarians and technicians have helped us to enroll over 45 farms that are distributed across several states, including Ohio, Indiana, Wisconsin, Kansas, and Michigan. Secondly, it has been difficult to gather movement records from state agencies. In many cases, legal exceptions exist for the requirement for Interstate Certificate of Veterinary Inspection, and Owner Shipper Statements are used instead. In some cases, these records are privately held, and are not accessible by state agencies. Therefore, we're working through how to manage the gaps in the data, and we will likely use some type of inferential method while being careful to avoid bias. Thirdly, although we have been able to enroll farms for the Salmonella Dublin component of this research, enrolling producers for quantification of AMU will be more difficult. We will need to work to directly engage producers and potentially provide monetary incentives so that producers will be willing to invest the time and energy necessary for completion of the objectives. What opportunities for training and professional development has the project provided?For the purposes of this project, we've hired two doctoral students, who have recently each successfully completed their candidacy examinations. Additionally, we've hired and trained three undergraduate students, that have worked in the laboratory and have garnered skills in microbiology and epidemiology. Lastly, two different post-doctoral researchers have collaborated with the faculty and students to design and execute the projects. The grant funding has allowed the faculty, post-doctoral researchers, and graduate students to attend the Conference of Research Workers in Animal Disease in Chicago, Illinois. This is a critical conference for developing researchers, and each of the trainees had an opportunity to present their work. How have the results been disseminated to communities of interest?We've disseminated our results to other academics and researchers through presentations at academic conferences. See the citations in the "Products" section. Additionally, we've begun work to disseminate general information and information from our research on biosecurity and antimicrobial use among dairy and calf producers. Firstly, we hosted a producer conference with approximatly 25 calf growers. The 6-hour conference was focused on Salmonella treatment, diagnosis, and prevention. Topics within the conferenceincluded introductory, producer-oriented information on antimicrobial resistance, practical considerations for biosecurity, working with the veterinarian for laboratory diagnostics, and research updates from our Team. Overall, the information and the conference was well-received by the producers. Secondly, our Team hosted booths at two producer dairy conferences, including the Northeast Ohio Dairy Symposium, and the Four Star Veterinary Clinic Symposium. At these conferences, we distributed fact sheets focused on biosecurity and S. Dublin,and had one-on-one interactions with producers that were very valuable for building relationships with the agricutural community and understanding the perspectives and attitudes of farmers on these topics. Furthermore, we wrote two articles in the Buckeye Dairy News, including one focused on biosecurity to preventS.Dublinand cleaning and disinfection for disease prevention. Additionally, we wrote an article inFarm and Dairyon general biosecurity measures. For this article, we focused on the emergent highly pathogenic influenza strain. What do you plan to do during the next reporting period to accomplish the goals?We intend to make progress on all three of the Aims of the project over the next reporting period. The first Specific Aim is focused on understanding the transmission patterns and prevention practices forSalmonellaDublin. Epidemiology of S. Dublin.We've worked with practicing veterinarians to collect environmental samples and biosecurity practices from over 45 different calf producers. We're continue to acrue farms to this dataset and we intend to analyze these data to identify management practices associated with PCR positivity. These data will be used to define which interventions can be instituted for prevention. Network of surplus calf movements. Understanding the movement patterns of surplus calves is critical towards identifying transmission pathways and prevention practices. Although we've already characterized the network of import and export calf movements for Ohio, we're expanding this effort to all 50 states. We've submitted data requests to the Departments of Agriculture, and we're working on the vast logistics necessary for gathering these data. The second Specific Aim is focused in improving responsible antimicrobial use among dairy-beef and veal calf raisers. Quantifying antimicrobial use on calf production units.We have begun to collaborate with a veal and dairy-beef production company to quantify antimicrobials within their production facility. As a first step, we need to validate efficient and scaleable methods for antimicrobial use quantification. These methods are well-developed for lactating cow dairy and finishing beef production systems, but the unique nature of dairy-beef and veal production systems require efforts towards validation. Specifically we'll determine if the use of pharmaceutical sales records can accurately quantify AMU relative to drug container collection methods. Subsequently, we will enroll dairy-beef and calf production units to quantify AMU and implement the benchmarking objectives as described in the original proposal.

Impacts
What was accomplished under these goals? The first Specific Aim is focused on understanding the transmission patterns and prevention practices forSalmonellaDublin. For the first full year of this project, we've focused primarily on the epidemiology and detection of Salmonella Dublin, and important cause of disease and mortality in calf production systems. Firstly, we identified the prevalence of Salmonella Dublin in special-fed veal calves, and identified this pathogen as a significant public health concern. Our research shows a notable presence of Salmonella Dublin in mesenteric lymph nodes, with variations in prevalence linked to geographic origin. This finding underscores the importance of location-specific strategies in managing Salmonella risks. For cattle producers, particularly those involved in veal production, these results highlight the need for targeted preharvest interventions. By understanding the prevalence and distribution of Salmonella Dublin, producers can implement more effective control measures, reducing the risk of infection and protecting consumer health. Secondly, our team developed a novel PCR forefficient detection ofSalmonella Dublin using multiplex PCR. S. Dublin is notoriously difficult to culture from environmental and fecal samples of subclinical shedders. Previously, there wasn't a viable technique for accurate detection in the environment.This technique proved more sensitive and accurate than traditional culture methods for identifying thepathogen in environmental samples. For cattle producers, the adoption of multiplex PCR will offer a valuable tool for early detection and monitoring of Salmonella Dublin in various cattle environments. This enhanced detection capability can lead to more timely interventions, reducing the spread of the pathogen and minimizing associated production losses. Moreover, this PCR will be a very important research to tool to better describe the epidemiology ofS.Dublin within cattle production systems. With the development of the novel test, we applied this assay to identify the prevalence and risk factors for Salmonella Dublin on dairy-beef farms. We collaboratored with veterinarians and veterinary technician that frequently visit these types of production facilities. We collected boot swabs from different areas of the farms, and developed a survey instrument to characterize the biosecurity practices on these farms. To date, we've collected samples and information from over 45 farms. These results have been presented at the Conference of Research Workers in Animal Disease. Additionally, we applied the assay to understand the environmental contamination of a calf dealer with Salmonella species and Salmonella Dublin. Approximately 25% of those environmental samples were positive for S. Dublin, highlighting the importance of auctions and calf dealers for dissemination of these pathogens. We've also made substantial progress towards characterization of the movement network of surplus calves from dairy farms. We collaborated with the Ohio Department of Agriculture to collect Interstate Certificates of Veterinary Inspection and Owner Shipper Statements to determine the network of calf import and exports to Ohio. Thisresearch emphasizes the critical role of comprehensive traceability systems in managing disease outbreaks. We found that relying solely on Interstate Certificates of Veterinary Inspection (ICVIs) underestimates the complexity and scope of calf movement networks. When Owner Shipper Statements (OSSs) were included, the network revealed a more extensive and cohesive structure. This suggests that current traceability methods may not fully capture all animal movements, which can obscure the true dynamics of disease transmission. For cattle producers, incorporating multiple data sources into traceability systems can significantly enhance the effectiveness of disease surveillance. This holistic approach can help prevent and manage outbreaks more efficiently, thereby protecting both animal and human health. This information is critical as we expand these efforts out to many states. Currently, we've requested comparable data from the Departments of Agriculture of nearly all 50 states. We hope to aggregate the available data to build one of the most comprehensive cattle movement networks to date. Overall, these research findings provide valuable insights for cattle producers aiming to improve disease management and biosecurity practices. By adopting comprehensive traceability systems, understanding the geographic prevalence of pathogens, and utilizing advanced detection methods, producers can better safeguard their herds and contribute to public health efforts.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Sequeira, S., Habing, G., Arruda, A. (2024). Combining multiple sources of movement data strengthens traceability and disease surveillance. In Proceedings of the Conference of Research Workers in Animal Disease. https://crwad.org/wp-content/uploads/CRWAD-2024-Proceedings-1.pdf
• Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Locke, S., Vinayamohan, P., Arevalo-Mayorga, A., Schaffner, V., Dunmyre, A., Ridolfo, J., Habing, G. (2024). Prevalence of Salmonella Dublin in mesenteric lymph nodes of special-fed veal calves from the US and Canada. In Proceedings of the 104th Conference of Research Workers in Animal Disease. https://crwad.org/wp-content/uploads/CRWAD-2024-Proceedings-1.pdf
• Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Arevalo-Mayorga, A., Ridolfo, J., Locke, S., Habing, G. (2024). Detection of Salmonella Dublin from environmental samples by end point multiplex PCR. In Proceedings of the 104th Conference of Research Workers in Animal Disease. https://crwad.org/wp-content/uploads/CRWAD-2024-Proceedings-1.pdf

Progress 05/01/22 to 04/30/23

Outputs
Target Audience:This research aims to understand the dissemination of S.Dublinand to optimize the use of antimicrobialswithin the surplus calf production systems. The target audience for the outputs of this grant proposal include those working in the surplus calf industry and researchers with similarly interested in the prevention and control of outbreaks ofSalmonellaDublin in calf populations and the prevention of transmission of antimicrobial resistant zoonotic pathogensthrough the food supply. Individuals working in the suplus calf industry includesthose that transport, sell, and auction calves. Additionally, the results will be of interest to calf growers, who often struggle with the health and welfare impacts ofS.Dublin, including veal, dairy-beef, and replacement heifer growers. More broadly, those who market dairy-beef and/or veal calves may interested in the impacts of improved antimicrobial stewardship on consumer product perception. Changes/Problems:We are addressing all the objectives originally described in the grant proposal. To broaden the impact of the research activities, we've made some important additions. Firstly, the team decidedto develop a PCR-based assay for detection ofS.Dublin in environmental samples. The availability of this assay addresses a key limitation in previousS.Dublin research, this is, the limited sensitivity of culture-based assays to detectS.Dublin. Secondly, the team decided to determine if slaughter-based sampling (lymph nodes) was an effective method to recoverS.Dublin in adult populations of dairy-beef cattle. Beyond these changes, we expect to accomplish the original Aims as written in the proposal. What opportunities for training and professional development has the project provided?We expect the project to provide high quality training opportunities for graduate student training in Microbiology, Epidemiology, and Molecular Epidemiology. Two doctoral students have been hired to work on the project and are actively working towards completion of the activities. One of the students will primarily focus on microbiological and molecular epidemiological aspects of the project, while another student will focus on the epidemiological aspects, including constructing a network of surplus calf movements and creating of model to prioritize interventions. Additionally, we've hired a post-doc that will oversee components of the study, including the cross-sectional study to estimate the prevalence of S. Dublin in the environment. Furthermore, there have been several undergraduate students experience research for the first time and assisting the graduate students with sample collection and microbiological procedures. How have the results been disseminated to communities of interest?Although it's early in the project, we've begun to dessiminate results to the target audiences. For instance, we're submitting three abstracts to the Conference of Research Workers in Animal Disease. These abstracts detail results related to the network of calf movements and the environmental prevalence ofS.?Dublin in surplus calf environments. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period, we're aiming to complete all activities within Specific Aim 1. This includes completion of sampling in the cross-sectional study of S. Dublin in the environment, characterization of S. Dublin recovered isolates, and data analysis to identify important risk factors. Additionaly activies include gathering ICVI and OSS data from state departments of agricutlure to broaden our description of surplus calf movements. Additionally, we will begin activities relating to the 2nd Aim of the project. Briefly, the overall goal of the 2nd aim of the project is to Identify interventions not reduce antimicrobial selection pressure in surplus calf facilities. More specifically, we're aiming to test the ability of benchmarking reports to reduce the quantity of antimicrobials used on farms. To conduct this farm-level randomized trial, we'll first meet with the leadership of one or more large production companies to explain the goals and objectives of the study. We'll also begin creating detailed study protocols and strategizing on how to enroll enough farms. We also need to determine the most efficient methods for quantifying antimicrobials given the unique nature of these production systems. Regardless, given our existing relationships with producers, we expect this component of the project to go smoothly.

Impacts
What was accomplished under these goals? The first year of the project was focused within the first Specific Aim 1. Briefly, the three objectives of the first Specific Aim were to 1) Describe the network of surplus calf movements in the Midwest and Eastern United States, 2)Characterize the genomic epidemiology of MDR Salmonella Dublin, and 3) Prioritize interventions for mitigating the dissemination of AMR pathogens. Objective 1: Characterize the network of surplus calf movements in the United States. To start with, we've worked directly with the Ohio Department of Agriculture to collect surplus calf movement records through Interstate Veterinary Certificates of Inspection (ICVIs) and Owner Shipper Statements (OSSs). This information was collected over a year timeframe and included all calf imports and exports to Ohio. This data was used to describe a preliminary network of calf movements that illustrates the geographic breadth. We'll expand this network by working with the Departments of Agriculture in states with extensive dairy industries, including Wisconsin, Pennsylvania, New York, and Indiana. We expect to begin broadening this network towards the end of this year. An additional part of the objective was to describe local calf movements not captured through ICVIs and OSSs. To accomplish this, we're working directly with a large calf dealer in Ohio, who is providing movement records and addresses so that an exemplary local network can be constructed. An abstract has been presented that describes the network of calf movements into and out of Ohio. Objective 2: Describe the genomic epidemiology of Salmonella Dublin. Our activities with regards to Objective 2 includes three studies: 1) Development of a S. Dublin specific PCR for detection of the organism in environmental samples, 2) A cross-sectional study of the prevalence if S. Dublin in fed dairy-beef cattle, and 3) A cross-sectional study of the prevalence of S. Dublin in the environments of dairy-beef and veal calf environments. For the first study, we developed a PCR specific to S. Dublin. Culture of S. Dublin from fecal and environmental samples is notoriously difficult; yet accurate detection from the environments of surplus calves is necessary to understand the transmission patterns of the organism. The PCR is specific to S. Dublin in-vitro and has a low limit of detection. Furthermore, we're comparing the results of PCR and culture in environmental samples from a calf dealer. So far, PCR has a much higher sensitivity relative to culture for detection of the organism. These are promising findings, and the availability of the test will result in new avenues for research. For the second study, we collaborated with a large slaughter plant in the Midwest that specialized in harvesting fed dairy-beef cattle. We hypothesized that dairy-beef cattle would have a relatively high prevalence of S. Dublin in the mesenteric lymph nodes. We collected 3-5 lymph nodes from > 600 cattle originating from several states and >50 different premises. Although Salmonella spp. were highly prevalent, none of the lymph nodes were positive for S. Dublin. This study will be useful to describe the prevalence, serotypes, and antimicrobial resistance of Salmonella spp. infecting dairy-beef cattle, and it's particularly interesting to note the very low prevalence in this population. This result is strikingly different from studies we've conducted in 5-month old veal calves, where at least half of the cohorts had at least one S. Dublin infected animal. Thirdly, we've begun a project to estimate the prevalence and identify risk factors for S. Dublin in environmental samples on dairy-beef and veal farms. To accomplish this, we're collaborating with a veterinary clinic with a large and geographically broad list of dairy-beef and clientele. We're using both culture and the S. Dublin specific PCR, and collecting potential risk factors (e.g. biosecurity practices) that may explain the presence of S. Dublin in the environmental samples. Additionally, we're collaborating with a large, vertically integrated veal calf production company and collecting similar samples and data. We expect to gather data from a large number of farms that will enable a precise prevalence estimate and identification of modifiable risk factors. At the moment, we're working to complete activities related to the first two objectives of the first Specific Aim and simultaneously planning activites for the latter part of Specific Aim 1 and the beginning of Specific Aim 2.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: S. Sequeira, G. Habing, A. G. Arruda. Description of calf production systems and network patterns to improve disease surveillance. 2023 College of Veterinary Medicine Research Day. April 2023, Columbus, OH