Analysis and modeling of Blastomyces dermatitidis environmental prevalence in Minnesota using soil collected to compare basal and outbreak levels.

Outbreaks of blastomycosis, caused by the fungus Blastomyces dermatitidis, occur in endemic areas of the United States and Canada but the geographic range of blastomycosis is expanding. Previous studies inferred the location of B. dermatitidis through epidemiologic data associated with outbreaks because culture of B. dermatitidis from the environment is often unsuccessful. In this study, we used a culture-independent, PCR-based method to identify B. dermatitidis DNA in environmental samples using the BAD1 promoter region. We tested 250 environmental samples collected in Minnesota, either associated with blastomycosis outbreaks or environmental samples collected from high- and low-endemic regions to determine basal prevalence of B. dermatitidis in the environment. We identified a fifth BAD1 promoter haplotype of B. dermatitidis prevalent in Minnesota. Ecological niche analysis identified latitude, longitude, elevation, and site classification as environmental parameters associated with the presence of B. dermatitidis Using this analysis, a Random Forest model predicted B. dermatitidis presence in basal environmental samples with 75% accuracy. These data support use of culture-independent, PCR-based environmental sampling to track spread into new regions and to characterize the unknown B. dermatitidis environmental niche.Importance Upon inhalation of spores from the fungus Blastomyces dermatitidis from the environment, humans and animals can develop the disease blastomycosis. Based on disease epidemiology, B. dermatitidis is known to be endemic in the United States and Canada around the Great Lakes and in the Ohio and Mississippi River Valleys but is starting to emerge in other areas. B. dermatitidis is extremely difficult to culture from the environment so little is known about the environmental reservoirs for this pathogen. We used a culture-independent PCR-based assay to identify the presence of B. dermatitidis DNA in soil samples from Minnesota. By combining molecular data with ecological niche modeling, we were able to predict the presence of B. dermatitidis in environmental samples with 75% accuracy and to define characteristics of the B. dermatitidis environmental niche. Importantly, we showed the effectiveness of using a PCR-based assay to identify B. dermatitidis in environmental samples.

Comparison of Antimicrobial-Resistant Escherichia coli Isolates from Urban Raccoons and Domestic Dogs.

Wildlife can be exposed to antimicrobial-resistant bacteria (ARB) via multiple pathways. Spatial overlap with domestic animals is a prominent exposure pathway. However, most studies of wildlife-domestic animal interfaces have focused on livestock and little is known about the wildlife-companion animal interface. Here, we investigated the prevalence and phylogenetic relatedness of extended-spectrum cephalosporin-resistant (ESC-R) Escherichia coli from raccoons (Procyon lotor) and domestic dogs (Canis lupus familiaris) in the metropolitan area of Chicago, IL, USA. To assess the potential importance of spatial overlap with dogs, we explored whether raccoons sampled at public parks (i.e., parks where people and dogs could enter) differed in prevalence and phylogenetic relatedness of ESC-R E. coli to raccoons sampled at private parks (i.e., parks where people and dogs could not enter). Raccoons had a significantly higher prevalence of ESC-R E. coli (56.9%) than dogs (16.5%). However, the richness of ESC-R E. coli did not vary by host species. Further, core single-nucleotide polymorphism (SNP)-based phylogenetic analyses revealed that isolates did not cluster by host species, and in some cases displayed a high degree of similarity (i.e., differed by less than 20 core SNPs). Spatial overlap analyses revealed that ESC-R E. coli were more likely to be isolated from raccoons at public parks than raccoons at private parks, but only for parks located in suburban areas of Chicago, not urban areas. That said, ESC-R E. coli isolated from raccoons did not genetically cluster by park of origin. Our findings suggest that domestic dogs and urban/suburban raccoons can have a diverse range of ARB, some of which display a high degree of genetic relatedness (i.e., differ by less than 20 core SNPs). Given the differences in prevalence, domestic dogs are unlikely to be an important source of exposure for mesocarnivores in urbanized areas. IMPORTANCE Antimicrobial-resistant bacteria (ARB) have been detected in numerous wildlife species across the globe, which may have important implications for human and animal health. Wildlife can be exposed to ARB via numerous pathways, including via spatial overlap with domestic animals. However, the interface with domestic animals has mostly been explored for livestock and little is known about the interface between wild animals and companion animals. Our work suggests that urban and suburban wildlife can have similar ARB to local domestic dogs, but local dogs are unlikely to be a direct source of exposure for urban-adapted wildlife. This finding is important because it underscores the need to incorporate wildlife into antimicrobial resistance surveillance efforts, and to investigate whether certain urban wildlife species could act as additional epidemiological pathways of exposure for companion animals, and indirectly for humans.

Lead Intoxication in Free-Ranging Bald Eagles ( Haliaeetus leucocephalus).

Lead toxicity due to ingestion of spent ammunition is an ongoing cause of mortality in bald eagles. While gross and histologic lesions of lead intoxication have been described in a few individuals of this species, the prevalence of lesions is underreported. A retrospective study of 93 bald eagles with severe lead intoxication was performed to describe the associated lesions and their prevalence and to compare the lesions with blood, liver, kidney, and/or bone lead concentrations. Gross lesions associated with lead toxicity were most frequent within the heart (51/93 birds) and consisted of multifocal myocardial pallor and rounding of the apex. Within the brain, gross lesions included petechiae or hemorrhagic necrosis (13/93 birds). Histologic lesions compatible with lead toxicity occurred within the heart (76/93 birds), brain (59/93 birds), and eyes (24/87 birds). Lead toxicity in bald eagles is characterized by fibrinoid necrosis of small- to medium-caliber arteries, most commonly affecting the heart, brain, and eyes. Gross and histologic lesions are consistent with ischemia caused by a primary vascular injury. A blood lead concentration of greater than 4 ppm and markedly elevated liver lead concentrations were associated with a greater likelihood of lesions in the heart. Severe lead intoxication is frequently associated with lesions that are histologically detectable in bald eagles. The presence of fibrinoid arterial necrosis and parenchymal degeneration, necrosis, and/or hemorrhage within the heart, brain, and/or eyes is suggestive of lead toxicity in bald eagles and warrants evaluation of liver or bone lead concentrations.

Live Animal Markets in Minnesota: A Potential Source for Emergence of Novel Influenza A Viruses and Interspecies Transmission.

BACKGROUND: Live animal markets have been implicated in transmission of influenza A viruses (IAVs) from animals to people. We sought to characterize IAVs at 2 live animal markets in Minnesota to assess potential routes of occupational exposure and risk for interspecies transmission. METHODS: We implemented surveillance for IAVs among employees, swine, and environment (air and surfaces) during a 12-week period (October 2012-January 2013) at 2 markets epidemiologically associated with persons with swine-origin IAV (variant) infections. Real-time reverse transcription polymerase chain reaction (rRT-PCR), viral culture, and whole-genome sequencing were performed on respiratory and environmental specimens, and serology on sera from employees at beginning and end of surveillance. RESULTS: Nasal swabs from 11 of 17 (65%) employees tested positive for IAVs by rRT-PCR; 7 employees tested positive on multiple occasions and 1 employee reported influenza-like illness. Eleven of 15 (73%) employees had baseline hemagglutination inhibition antibody titers ≥40 to swine-origin IAVs, but only 1 demonstrated a 4-fold titer increase to both swine-origin and pandemic A/Mexico/4108/2009 IAVs. IAVs were isolated from swine (72/84), air (30/45), and pen railings (5/21). Whole-genome sequencing of 122 IAVs isolated from swine and environmental specimens revealed multiple strains and subtype codetections. Multiple gene segment exchanges among and within subtypes were observed, resulting in new genetic constellations and reassortant viruses. Genetic sequence similarities of 99%-100% among IAVs of 1 market customer and swine indicated interspecies transmission. CONCLUSIONS: At markets where swine and persons are in close contact, swine-origin IAVs are prevalent and potentially provide conditions for novel IAV emergence.

Validation of good agricultural practices (GAP) on Minnesota vegetable farms.

The United States Food and Drug Administration and the Department of Agriculture jointly published the "Guide to Minimize Microbial Food Safety Hazards for Fresh Fruits and Vegetables," which is used as a basis for Good Agricultural Practices (GAP) audits. To understand barriers to incorporation of GAP by Minnesota vegetable farmers, a mail survey completed in 2008 was validated with visits to a subset of the farms. This was done to determine the extent to which actual practices matched perceived practices. Two hundred forty-six producers completed the mail survey, and 27 participated in the on-farm survey. Over 75% of the on-farm survey respondents produced vegetables on 10 acres or less and had 10 or fewer employees. Of 14 questions, excellent agreement between on-farm interviews and mail survey responses was observed on two questions, four questions had poor or slight agreement, and eight questions had no agreement. Ninety-two percent of respondents by mail said "they took measures to keep animals and pests out of packing and storage buildings." However, with the on-site visit only 45% met this requirement. Similarly, 81% of respondents by mail said "measures were taken to reduce the risk of wild and/or domestic animals entering into fruit and vegetable growing areas." With direct observation, 70% of farms actually had taken measures to keep animals out of the growing areas. Additional, on-farm assessments were done regarding employee hygiene, training, presence of animals, water sources, and composting practices. This validation study demonstrated the challenge of creating nonleading and concise questions that are not open to broad interpretation from the respondents. If mail surveys are used to assess GAP, they should include open-ended questions and ranking systems to better assess farm practices. To provide the most accurate survey data for educational purposes or GAP audits, on-farm visits are recommended.

An increase in animal-related occupational injuries at a veterinary medical center (2008-2022).

OBJECTIVE: To characterize animal-related injuries in veterinary medical center staff at a veterinary medical center. SAMPLE: 706 hospital staff injuries. METHODS: Deidentified injury reports were submitted to Human Resources from 2008 through 2022. Injury data collected included the injury description, date of injury, occupation, and worker's compensation claim information. Data were summarized by year, cause of injury, total cost associated with injury, and occupation. RESULTS: There was an increase in injuries reported in recent years when compared to past years, with the plurality of injuries being bite injuries, specifically occurring on the hand, finger, and wrist area. Bite injuries had a higher average total worker's compensation cost paid to staff than striking injuries. There were more injuries reported by staff who had less experience working with animals. More injuries occurred during the summer months (June through September). There was not an unusual trend in the reporting of injuries due to COVID-19. Other injuries (eg, needlesticks and falls) were reported from only 2019 to 2022, but constituted a substantial burden for staff. CLINICAL RELEVANCE: These findings can help stakeholders at teaching hospitals and veterinary clinics to take steps toward creating a safer workplace environment for employees. It is important to identify work hazards and provide proper training and prevention methods to reduce the risk of injuries, especially among less experienced employees. Proper prevention methods will help reduce worker's compensation costs for the teaching hospital and reduce the number of workdays missed by staff.

It Takes a Village: A Novel Process for Responding to Emerging Issues in Agricultural Health and Safety.

Agricultural and food production systems are constantly evolving in response to societal and environmental factors. These sectors are also laden with occupational hazards, creating an opportunity for emerging and re-emerging issues such as emerging markets and changing worker demographics. The Upper Midwest is a region of intensive agricultural production, with many states leading the United States in livestock and crop production. As a National Institute of Occupational Safety and Health Center for Agricultural Safety and Health, the Upper Midwest Agricultural Safety and Health Center (UMASH) is tasked with protecting the health and safety of the region's agricultural workers. UMASH implements an Emerging Issues program to effectively respond to emerging and re-emerging issues impacting agricultural health and safety in the Upper Midwest. To direct such work, UMASH developed a novel Selection, Planning and Action Process Model. This model guides all stages of the UMASH Emerging Issues response and emphasizes the ongoing need for monitoring, evaluation, and feedback from agricultural communities. In conjunction with the UMASH Process Model, the International Association for Public Participation (IAP2) Spectrum of Public Participation frames the work of the Emerging Issues Program in meaningfully engaging the community of stakeholders across all stages.

Assessing Self-reported Occupational Hazards of Manure Applicators in the Upper Midwest.

Manure management on farms presents potential risks to human health and safety, including infectious, chemical, and physical exposures that may result in injury or fatality. Toxic gases and confined spaces are among the most common hazards. These hazards are especially salient for the Upper Midwest of the United States. This study characterizes the occupational health and safety practices and experiences of manure applicators in Minnesota. This cross-sectional study surveyed 162 commercial manure applicators about their work characteristics, safety practices, and health and safety experiences. Respondents reported an average of 17 hours per day applying manure during the busy season, which typically occurs several weeks each year. One hundred and thirty-one (90%) of 145 respondents did not regularly use gas monitors during application. Thirty-three (37%) of 90 respondents reported at least one symptom of gas exposure during manure application work. Those that worked with swine manure were more likely to report symptoms of gas exposure than those who did not work with swine (OR 9.5; CI: 2.0, 89.0). Those that had entered confined spaces were more likely to report symptoms of gas exposure than those who had not entered confined spaces (OR 4.4; CI: 0.1, 1.1). Fourteen (9%) of 150 respondents reported being injured when applying manure. Manure work can be hazardous and may be associated with injury and gas exposure. These findings offer a starting point for future research and intervention to protect and improve the health and safety of applicators.

The potential capability of social media as a component of food safety and food terrorism surveillance systems.

Social media (i.e., internet applications and platforms that allow users to create and share content) are a potentially rich data source for foodborne disease surveillance. This commentary qualitatively evaluates social media as a foodborne disease surveillance system component and discusses novel ways that social media may facilitate the detection of intentional and unintentional food contamination.

A longitudinal study of Salmonella from snakes used in a public outreach program.

Snakes are considered to be a source of Salmonella infection for humans, but little is known about the actual serotype prevalence in healthy snakes over time. Twelve snakes involved in a public outreach program, representing seven different species, were tested weekly for shedding of Salmonella sp. over a period of 10 consecutive weeks. The snakes were housed in close proximity but in separate exhibits. Fresh fecal samples (when available) or cloacal swabs were cultured for Salmonella sp., and subsequent Salmonella isolates were serotyped. As representatives of the feed source, the feces of two mice and the intestines of one rat were cultured weekly. Fecal samples from 11 of the 12 snakes were positive for Salmonella at least once. Seven (58%) of 12 snakes were culture positive five times or more. The weekly prevalence of Salmonella shedding varied between 25% and 66%. Two or more different serotypes were isolated from nine snakes over time; however, a predominant serotype was generally isolated from each of these snakes. Altogether 15 different serotypes were identified. Serotypes of public health concern included Newport, Oranienburg, and Muenchen. Two samples from feeder rodents were positive for Salmonella. The results are consistent with previous studies showing high intestinal colonization rates with Salmonella sp. in snakes. Frequent and intermittent shedding of multiple serotypes was evident. Feeder rodents might serve as a source for intestinal colonization. Appropriate handling protocols should be implemented for all reptiles associated with public outreach programs to minimize risk of Salmonella transmission to the public.

Global Distribution of Extended Spectrum Cephalosporin and Carbapenem Resistance and Associated Resistance Markers in Escherichia coli of Swine Origin - A Systematic Review and Meta-Analysis.

Third generation cephalosporins and carbapenems are considered critically important antimicrobials in human medicine. Food animals such as swine can act as reservoirs of antimicrobial resistance (AMR) genes/bacteria resistant to these antimicrobial classes, and potential dissemination of AMR genes or resistant bacteria from pigs to humans is an ongoing public health threat. The objectives of this systematic review and meta-analysis were to: (1) estimate global proportion and animal-level prevalence of swine E. coli phenotypically resistant to third generation cephalosporins (3GCs) and carbapenems at a country level; and (2) measure abundances and global distribution of the genetic mechanisms that confer resistance to these antimicrobial classes in these E. coli isolates. Articles from four databases (CAB Abstracts, PubMed/MEDLINE, PubAg, and Web of Science) were screened to extract relevant data. Overall, proportion of E. coli resistant to 3GCs was lower in Australia, Europe, and North America compared to Asian countries. Globally, <5% of all E. coli were carbapenem-resistant. Fecal carriage rates (animal-level prevalence) were consistently manifold higher as compared to pooled proportion of resistance in E. coli isolates. bla CTX-M were the most common 3GC resistance genes globally, with the exception of North America where bla CMY were the predominant 3GC resistance genes. There was not a single dominant bla CTX-M gene subtype globally and several bla CTX-M subtypes were dominant depending on the continent. A wide variety of carbapenem-resistance genes (bla NDM-, VIM-, IMP-, OXA-48, and KPC-) were identified to be circulating in pig populations globally, albeit at very-low frequencies. However, great statistical heterogeneity and a critical lack of metadata hinders the true estimation of prevalence of phenotypic and genotypic resistance to these antimicrobials. Comparatively frequent occurrence of 3GC resistance and emergence of carbapenem resistance in certain countries underline the urgent need for improved AMR surveillance in swine production systems in these countries.

Occurrence and potential transmission of extended-spectrum beta-lactamase-producing extraintestinal pathogenic and enteropathogenic Escherichia coli in domestic dog faeces from Minnesota.

Interactions between humans and pets are increasingly valued in western countries, leading to more extensive contact between humans and their pets within households. Although the magnitude of the risk of transfer of Escherichia coli between humans and their companion animals is undefined, that such transmission occurs has been established and warrants attention. This study examined 186 fresh faecal samples from companion dogs visiting 22 municipal dog parks in the Minneapolis/Saint Paul metropolitan area, Minnesota, USA. Samples were processed to isolate 3rd-generation cephalosporin-resistant E. coli, which were further characterized using PCR-based virulence genotyping, antimicrobial susceptibility profiling and whole-genome sequencing. Of the 186 faecal samples, 29% yielded cephalosporin-resistant E. coli, and 2.2% yielded extended-spectrum beta-lactamase producers. Co-resistance to sulfonamides was typical (77.3% of isolates), and multidrug resistance (i.e. to ≥3 antimicrobial classes), including to combinations of tetracyclines, phenicols, quinolones and aminoglycosides, was substantial (18.9% of isolates). Identified beta-lactamase genes included blaCMY-2 , blaTEM-1B , blaTEM-1 , blaCTX-M-24 , blaCTX-M-15 and blaOXA-1 . Genome sequencing of 14 isolates identified genes typical of extraintestinal pathogenic E. coli or enteropathogenic E. coli. In three instances, closely related isolates were recovered from different dogs, within either the same park-suggesting transfer of E. coli between dogs within the park-or different parks-suggesting that dogs may be pre-disposed to carry certain E. coli types, such as those from serogroups O4, O71 and O157. This study adds to the existing evidence that companion dogs can harbour and share antimicrobial-resistant E. coli with presumed intestinal or extraintestinal pathogenic potential.

Global Distribution of Fluoroquinolone and Colistin Resistance and Associated Resistance Markers in Escherichia coli of Swine Origin - A Systematic Review and Meta-Analysis.

Background: Fluoroquinolones and polymyxins (colistin) are considered as critical drugs for human medicine. Antimicrobials of these classes are also used in swine production worldwide and this usage can contribute to selection of antimicrobial resistance (AMR), which is a threat to both human and animal health. Given the dynamic epidemiology of AMR, updating our knowledge regarding distribution and trends in the proportion of resistant bacteria is of critical importance. Objectives: The aim of this systematic review and meta-analysis was to describe the global prevalence of phenotypic and genotypic resistance to fluoroquinolones and colistin in Escherichia coli collected from swine. Results: Four databases (PubMed, PubAg, Web of Science, and CAB abstracts) and reports of national surveillance programs were scanned and 360 articles were included in the analysis. We identified higher prevalence levels of fluoroquinolone and colistin resistance in isolates from pig populations in Asia compared to Europe. The heterogeneity of pooled estimates was also higher in Asian countries suggesting that prevalence of AMR is still not fully characterized. There was a major knowledge gap about the situation of AMR in South American and African countries. We also identified key deficiencies in how AMR data was reported in the studies. A meta-analysis using 6,167 publicly available genomes of swine E. coli established the prevalence and global distribution of genetic determinants that can lead to fluoroquinolone and colistin resistance. Conclusion: This study provides the most comprehensive information on prevalence of phenotypic and genotypic resistance to key antimicrobials in pig populations globally. There is a need to establish national surveillance programs and effective policies, particularly in certain world regions, to curtail the threat of evolution of resistant isolates in swine production that can potentially contribute to public health detrimentally.

Strengthening global health security by improving disease surveillance in remote rural areas of low-income and middle-income countries.

The COVID-19 pandemic has underscored the need to strengthen national surveillance systems to protect a globally connected world. In low-income and middle-income countries, zoonotic disease surveillance has advanced considerably in the past two decades. However, surveillance efforts often prioritise urban and adjacent rural communities. Communities in remote rural areas have had far less support despite having routine exposure to zoonotic diseases due to frequent contact with domestic and wild animals, and restricted access to health care. Limited disease surveillance in remote rural areas is a crucial gap in global health security. Although this point has been made in the past, practical solutions on how to implement surveillance efficiently in these resource-limited and logistically challenging settings have yet to be discussed. We highlight why investing in disease surveillance in remote rural areas of low-income and middle-income countries will benefit the global community and review current approaches. Using semi-arid regions in Kenya as a case study, we provide a practical approach by which surveillance in remote rural areas can be strengthened and integrated into existing systems. This Viewpoint represents a transition from simply highlighting the need for a more holistic approach to disease surveillance to a solid plan for how this outcome might be achieved.

Establishing a milkborne disease outbreak profile: potential food defense implications.

OBJECTIVES: The main objectives of this study were to establish baseline characteristics for milkborne outbreaks, establish an expected milkborne outbreak profile, and identify potential indicators of food terrorism. MATERIALS: This study used 1990-2006 data from the Centers for Disease Control and Prevention Annual Listings of Disease Outbreaks and the Foodborne Outbreak Database (FOOD) to establish epidemiologic baseline characteristics for disease outbreaks associated with fluid milk. FOOD data from 2007 were used to qualitatively validate the potential of the baseline characteristics and the expected outbreak profile. RESULTS: Eighty-three fluid milkborne outbreaks were reported between 1990 and 2006, resulting in 3621 illnesses. The mean number of illnesses per outbreak was 43.6 (illness range: 2-1644). Consumption of unpasteurized milk was associated with 55.4% of reported outbreaks. Campylobacter spp., Escherichia coli, and Salmonella spp. caused 51.2%, 10.8%, and 9.6% of reported outbreaks, respectively. Private homes accounted for 41.0% of outbreak locations. Number ill, outbreak location, and etiology were the primary characteristics which could signal a potential intentional contamination event. In 2007, one pasteurized milk outbreak caused by Listeria was flagged as aberrative compared with the expected outbreak profile. CONCLUSIONS: The creation and dissemination of expected outbreak profiles and epidemiologic baseline characteristics allow public health and Homeland Security officials to quickly assess the potential of intentional food contamination. A faster public health and medical system response can result in decreased morbidity and mortality.

Developing an open-access antimicrobial resistance learning site for veterinary medical students.

Recognizing the crucial role of veterinarians in mitigating antimicrobial resistance (AMR), the Centers for Disease Control and Prevention (CDC) has funded the development of a suite of educational materials to promote the responsible veterinary medical use of antimicrobials. An open-access, Web-based multimedia curriculum regarding antimicrobial resistance in veterinary practice was thus created. The antimicrobial-resistance learning site (AMRLS) for veterinary medical students was completed and made available for use in January 2011 (http://amrls.cvm.msu.edu/). Designed for integration into existing veterinary medical courses, the AMRLS is also a resource for continuing education for practicing veterinarians, animal scientists, and food-animal industry specialists. This Web site emphasizes the mechanisms by which AMR emerges and spreads, the significant role of veterinarians in mitigating AMR, and the need to preserve the efficacy of antibiotics for future generations.

The SCCmec Types and Antimicrobial Resistance among Methicillin-Resistant Staphylococcus Species Isolated from Dogs with Superficial Pyoderma.

This study characterizes clinical methicillin-resistant staphylococcal (MRS) isolates obtained from superficial pyoderma infections in dogs. Our interest was to determine the staphylococcal cassette chromosome mec (SCCmec) type and the antimicrobial susceptibility among MRS isolates from clinical cases. Skin swabs were collected and cultured. Staphylococcus species were identified and characterized with biochemical tests and MALDI-TOF-MS and antimicrobial susceptibility testing by disk diffusion. mecA detection and staphylococcal cassette chromosome mec (SCCmec) typing were achieved by PCR. Of the 65 clinical samples, 56 (86.2%) staphylococcal infections were identified. Twelve (21%) of 56 isolates were MRS infections. All MRS isolates were multidrug resistant. The ccrC and class-C2 mec, which were SCCmec type V, were the most prevalent (66.7%) among the 12 MRS isolates. The predominant SCCmec type V was found in S. aureus, S. intermedius group, S. lentus, S. xylosus, and S. arlettae. Treatment failure is a concern with the emergence of highly resistant MRS in dogs associated with superficial pyoderma. The detection of type V SCCmec MRS has previously been reported among veterinarians and dog owners but not in Northern Thailand. These infections serve as a reminder to improve infection prevention and control measures including reducing environmental contamination and potential zoonotic exposures to MRS. In addition, educational awareness of these risks in small animal hospitals needs to be increased among veterinary hospital staff, clients, and patients.

COVID-19 Awareness and Preparedness of Minnesota and Wisconsin Dairy Farms.

Dairy farms that had participated in previous and ongoing projects with the National Farm Medicine Center (NFMC), Migrant Clinicians Network (MCN), and Upper Midwest Agricultural Safety and Health Center (UMASH) were asked to participate in a 17-question survey by phone or email to investigate biosecurity principles on Minnesota and Wisconsin dairy farms in response to COVID-19 and the effects of the pandemic on the dairy industry. Three additional farms were recruited via a press release published in agricultural newsletters. Of 76 farms contacted, 37 chose to participate in this study from June to July 2020. In response to the COVID-19 pandemic, dairies have implemented or increased biosecurity measures and COVID-19 precautions. Dairies reported adequate personal protective equipment for their workers, though face masks were not required on most dairies (n = 32, 86%). Producers were concerned about the safety of their families, maintaining a healthy workforce, and keeping their farms profitable. Access to healthcare was not perceived to be an issue for their workers. One-quarter of dairies reported COVID-19 infections on their farms. Even though the majority had an isolation protocol in place if someone on the farm were to become ill, less than half of respondents felt their farm was protected against COVID-19. Two-thirds of producers have not had to decrease production, and a majority of operations have not furloughed or terminated employees due to COVID-19. Our data suggest that dairy farms in Minnesota and Wisconsin have implemented biosecurity and safety measures in response to COVID-19. These measures can be improved. Farms would benefit from additional guidance and education on implementation of personal protective measures and disease prevention strategies to keep workers employed and safe.

Importance of anthropogenic sources at shaping the antimicrobial resistance profile of a peri-urban mesocarnivore.

Anthropogenically derived antimicrobial-resistant bacteria (ARB) and antimicrobial resistance genes (ARG) have been detected in wildlife. The likelihood of detecting ARB and ARG in wildlife increases with wildlife exposure to anthropogenic sources of antimicrobial resistance (AMR). Whether anthropogenic sources also increase the risk for AMR to spread in bacteria of wildlife is not well understood. The spread of AMR in bacteria of wildlife can be estimated by examining the richness of ARB and ARG, and the prevalence of ARB that have mobilizable ARG (i.e., ARG that can be transferred across bacteria via plasmids). Here, we investigated whether raccoons (Procyon lotor), with different exposures to anthropogenic sources, differed in prevalence and richness of extended-spectrum cephalosporin-resistant (ESC-R) Escherichia coli, richness of ARG present in ESC-R E. coli, and prevalence of ESC-R E. coli with plasmid-associated ARG. Sampling took place over the course of 10 months at seven sites in Chicago, USA. ESC-R E. coli were isolated from over half of the 211 raccoons sampled and were more likely to be isolated from urban than suburban raccoons. When examining the whole-genome sequences of ESC-R E. coli, 56 sequence types were identified, most of which were associated with the ARG blaCMY and blaCTX-M. A greater richness of ESC-R E. coli sequence types was found at sites with a wastewater treatment plant (WWTP) than without, but no difference was detected based on urban context. ARG richness in ESC-R E. coli did not significantly vary by urban context nor with presence of a WWTP. Importantly, ESC-R E. coli carrying plasmid-associated blaCTX-M and blaCMY ARG were more likely to be isolated from raccoons sampled at sites with a WWTP than without. Our findings indicate that anthropogenic sources may shape the AMR profile of wildlife, reinforcing the need to prevent dissemination of AMR into the environment.

Genetic diversity of influenza A viruses circulating in pigs between winter and summer in a Minnesota live animal market.

There has been little surveillance of influenza A viruses (IAVs) circulating in swine at live animal markets, particularly in the United States. To address this gap, we conducted active surveillance of IAVs in pigs, the air, and the environment during a summer and winter season in a live animal market in St. Paul, Minnesota, that had been epidemiologically associated with swine-origin influenza cases in humans previously. High rates of IAV were detected by PCR in swine lungs and oral fluids during both summer and winter seasons. Rates of IAV detection by PCR in the air were similar during summer and winter, although rates of successful virus isolation in the air were lower during summer than in winter (26% and 67%, respectively). H3N2 was the most prevalent subtype in both seasons, followed by H1N2. Genetically diverse viruses with multiple gene constellations were isolated from both winter and summer, with a total of 19 distinct genotypes identified. Comparative phylogenetic analysis of all eight segments of 40 virus isolates from summer and 122 isolates from winter revealed that the summer and winter isolates were genetically distinct, indicating IAVs are not maintained in the market, but rather are re-introduced, likely from commercial swine. These findings highlight the extent of IAV genetic diversity circulating in swine in live animal markets, even during summer months, and the ongoing risk to humans.