Antimicrobial use policy change in pre-weaned dairy calves and its impact on antimicrobial resistance in commensal Escherichia coli: a cross sectional and ecological study.

BACKGROUND: This study is based on data collected to investigate the relation of peri-parturient events (colostrum quality, passive transfer of immunity, calving difficulty) on calf health and antimicrobial use. A component of the study was to provide feedback to farm management to identify calves at risk for disease and promote antimicrobial stewardship. At the start of the study (May 2016), a combination of enrofloxacin, penicillin, and sulfamethoxazole was the first treatment given to clinically abnormal calves. Based on feedback and interaction between study investigators, farm management and consulting veterinarians, a new policy was implemented to reduce antimicrobial use in calves. In August, the first treatment was changed to a combination of ampicillin and sulfamethoxazole. In September, the first treatment was reduced to only sulfamethoxazole. We investigated the effects of these policy changes in antimicrobial use on resistance in commensal Escherichia coli. RESULTS: We enrolled 4301 calves at birth and documented antimicrobial use until weaning. Most calves (99.4%) received antimicrobials and 70.4% received a total of 2-4 treatments. Antimicrobial use was more intense in younger calves (≤ 28 days) relative to older calves. We isolated 544 E. coli from fecal samples obtained from 132 calves. We determined resistance to 12 antimicrobials and 85% of the isolates were resistant to at least 3 antimicrobial classes. We performed latent class analysis to identify underlying unique classes where isolates shared resistance patterns and selected a solution with 4 classes. The least resistant class had isolates that were mainly resistant to only tetracycline and sulfisoxazole. The other 3 classes comprised isolates with resistance to ampicillin, chloramphenicol, aminoglycosides, sulfonamides, tetracycline, in addition to either ceftiofur; or nalidixic acid; or ciprofloxacin plus nalidixic acid and ceftiofur. Overall, E coli from younger calves and calves that received multiple treatments were more likely to have extensive resistance including resistance to fluoroquinolones and ceftiofur. In general, there was a declining trend in resistance to most antimicrobials during and after policy changes were implemented, except for ampicillin, ciprofloxacin, ceftiofur and gentamicin. CONCLUSIONS: Information feedback to farms can influence farm managers to reduce antimicrobial use and this can change endemic farm resistance patterns.

Molecular Epidemiology of Dairy Cattle-Associated Escherichia coli Carrying blaCTX-M Genes in Washington State.

An increase in the prevalence of commensal Escherichia coli carrying blaCTX-M genes among dairy cattle was observed between 2008 and 2012 in Washington State. To study the molecular epidemiology of this change, we selected 126 blaCTX-M-positive and 126 blaCTX-M-negative isolates for determinations of the multilocus sequence types (MLSTs) and antibiotic resistance phenotypes from E. coli obtained during a previous study. For 99 isolates, we also determined the blaCTX-M alleles using PCR and sequencing and identified the replicon types of blaCTX-M-carrying plasmids. The blaCTX-M-negative E. coli isolates comprised 76 sequence types (STs) compared with 32 STs in blaCTX-M-positive E. coli isolates. The blaCTX-M-positive E. coli isolates formed three MLST clonal complexes, accounting for 83% of these isolates; 52% of blaCTX-M-negative E. coli isolates clustered into 10 clonal complexes, and the remainder were singletons. Overall, blaCTX-M-negative E. coli isolates had more diverse genotypes that were distinct to farms, whereas blaCTX-M-positive E. coli isolates had a clonal population structure and were widely disseminated on farms in both regions included in the study. Plasmid replicon types included IncI1 which predominated, followed by IncFIB and IncFIA/FIB. blaCTX-M-15 was the predominant CTX-M gene allele, followed by blaCTX-M-27 and blaCTX-M-14 There was no significant association between plasmid replicon types and bacterial STs, and neither clonal complexes nor major plasmid groups were associated with two discrete dairy-farming regions of Washington State.IMPORTANCE Infections caused by extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli occur globally and present treatment challenges because of their resistance to multiple antimicrobial drugs. Cattle are potential reservoirs of ESBL-producing Enterobacteriaceae, and so understanding the causes of successful dissemination of blaCTX-M genes in commensal bacteria will inform future approaches for the prevention of antibiotic-resistant pathogen emergence.

Whole-Genome Sequencing of Drug-Resistant Salmonella enterica Isolates from Dairy Cattle and Humans in New York and Washington States Reveals Source and Geographic Associations.

Multidrug-resistant (MDR) Salmonella enterica can be spread from cattle to humans through direct contact with animals shedding Salmonella as well as through the food chain, making MDR Salmonella a serious threat to human health. The objective of this study was to use whole-genome sequencing to compare antimicrobial-resistant (AMR) Salmonella enterica serovars Typhimurium, Newport, and Dublin isolated from dairy cattle and humans in Washington State and New York State at the genotypic and phenotypic levels. A total of 90 isolates were selected for the study (37 S Typhimurium, 32 S Newport, and 21 S Dublin isolates). All isolates were tested for phenotypic antibiotic resistance to 12 drugs using Kirby-Bauer disk diffusion. AMR genes were detected in the assembled genome of each isolate using nucleotide BLAST and ARG-ANNOT. Genotypic prediction of phenotypic resistance resulted in a mean sensitivity of 97.2 and specificity of 85.2. Sulfamethoxazole-trimethoprim resistance was observed only in human isolates (P < 0.05), while resistance to quinolones and fluoroquinolones was observed only in 6 S Typhimurium isolates from humans in Washington State. S Newport isolates showed a high degree of AMR profile similarity, regardless of source. S Dublin isolates from New York State differed from those from Washington State based on the presence/absence of plasmid replicons, as well as phenotypic AMR susceptibility/nonsusceptibility (P < 0.05). The results of this study suggest that distinct factors may contribute to the emergence and dispersal of AMR S. enterica in humans and farm animals in different regions.IMPORTANCE The use of antibiotics in food-producing animals has been hypothesized to select for AMR Salmonella enterica and associated AMR determinants, which can be transferred to humans through different routes. Previous studies have sought to assess the degree to which AMR livestock- and human-associated Salmonella strains overlap, as well as the spatial distribution of Salmonella's associated AMR determinants, but have often been limited by the degree of resolution at which isolates can be compared. Here, a comparative genomics study of livestock- and human-associated Salmonella strains from different regions of the United States shows that while many AMR genes and phenotypes were confined to human isolates, overlaps between the resistomes of bovine and human-associated Salmonella isolates were observed on numerous occasions, particularly for S Newport. We have also shown that whole-genome sequencing can be used to reliably predict phenotypic resistance across Salmonella isolated from bovine sources.

Comparative composition, diversity, and abundance of oligosaccharides in early lactation milk from commercial dairy and beef cows.

Prebiotics are nondigestible dietary ingredients, usually oligosaccharides (OS), that provide a health benefit to the host by directly modulating the gut microbiota. Although there is some information describing OS content in dairy-source milk, no information is available to describe the OS content of beef-source milk. Given the different trait emphasis between dairy and beef for milk production and calf survivability, it is plausible that OS composition, diversity, and abundance differ between production types. The goal of this study was to compare OS in milk from commercial dairy and beef cows in early lactation. Early-lactation multiparous cows (5-12 d in milk) from 5 commercial Holstein dairy herds and 5 Angus or Angus hybrid beef herds were sampled once. Milk was obtained from each enrolled cow and frozen on the farm. Subsequently, each milk sample was assessed for total solids, pH, and OS content and relative abundance. Oligosaccharide diversity and abundance within and between samples was transformed through principal component analysis to reduce data complexity. Factors from principal component analysis were used to create similarity clusters, which were subsequently used in a multivariate logistic regression. In total, 30 OS were identified in early-lactation cow milk, including 21 distinct OS and 9 isomers with unique retention times. The majority of OS detected in the milk samples were present in all individual samples regardless of production type. Two clusters described distribution patterns of OS for the study sample; when median OS abundance was compared between the 2 clusters, we found that overall OS relative abundance was consistently greater in the cluster dominated by beef cows. For several of the structures, including those with known prebiotic effect, the difference in abundance was 2- to 4-fold greater in the beef-dominated cluster. Assuming that beef OS content in milk is the gold standard for cattle, it is likely that preweaning dairy calves are deprived of dietary-source OS. Although supplementing rations with OS is an approach to rectify this deficiency, understanding the health and productivity effects of improving OS abundance being fed to preweaning calves is a necessary next step before recommending supplementation. These studies should account for the observation that OS products are variable for both OS diversity and structural complexity, and some products may not be suitable as prebiotics.

Improving Dairy Organizational Communication from the Veterinarian's Perspective: Results of a Continuing Veterinary Medical Education Pilot Program.

The increasing size and complexity of US dairy farms could make it more difficult for a veterinary practitioner to effectively communicate protocol recommendations for prevention or treatment on the farm. A continuing education workshop was set up based on the results of research on dairy organizational communication on dairy farms, which resulted in a tool to assess dairy communication structure and flow. The workshop specifically focused on communication structure and whom to talk to when implementing health care changes in calf rearing. In addition, modern methods of veterinary-client communication knowledge and skills were provided. Primary outcomes of the workshops were to obtain feedback from participants about research findings and the communication model, to improve awareness about the complexity of communication structures on dairy farms, and to change participants' knowledge and skills associated with on-farm communication by providing communication theory and skills and an approach to evaluate and improve dairy organizational communication. Of the 37 participants completing the pre-program assessment, most recognized a need for themselves or their practice to improve communication with clients and farm employees. After the program, most participants were confident in their new communication skills and would consider using them. They highlighted specific new ideas they could apply in practice, such as conducting a "communication audit." The results from the assessment of this communication workshop, focused on dairy veterinarians, highlighted the need for communication training in this sector of the profession and practitioners' desire to engage in this type of training.

Recent Emergence of Escherichia coli with Cephalosporin Resistance Conferred by blaCTX-M on Washington State Dairy Farms.

Enterobacteriaceae-associated blaCTX-M genes have become globally widespread within the past 30 years. Among isolates from Washington State cattle, Escherichia coli strains carrying blaCTX-M (CTX-M E. coli strains) were absent from a set of 2008 isolates but present in a set of isolates from 2011. On 30 Washington State dairy farms sampled in 2012, CTX-M E. coli prevalence was significantly higher on eastern than on northwestern Washington farms, on farms with more than 3,000 adult cows, and on farms that recently received new animals. The addition of fresh bedding to calf hutches at least weekly and use of residual fly sprays were associated with lower prevalence of CTX-M E. coli. In Washington State, the occurrence of human pathogens carrying blaCTX-M genes preceded the emergence of blaCTX-M-associated E. coli in cattle, indicating that these resistance determinants and/or their bacterial hosts may have emerged in human populations prior to their dissemination to cattle populations.

Contrasting Fecal Methanogenic and Bacterial Profiles of Organic Dairy Cows Located in Northwest Washington Receiving Either a Mixed Diet of Pasture and TMR or Solely TMR.

Currently, little is known regarding fecal microbial populations and their associations with methanogenic archaea in pasture-based dairy cattle. In this study, we assessed the fecal microbiome of organic dairy cows across different time points receiving a mixed diet of pasture and total mixed ration (TMR) or TMR only. We hypothesized that the fecal methanogenic community, as well as co-occurrence patterns with bacteria, change across diets. To test these hypotheses, we analyzed TMR and pasture samples, as well as the V3-V4 region of 16S rRNA of fecal samples collected over the course of a one-year study period from 209 cows located on an organic dairy in Northwest Washington. The inherent variability in pasture quality, quantity, availability, and animal preference can lead to diverse dietary intakes. Therefore, we conducted a k-means clustering analysis to identify samples from cows that were associated with either a pasture-based diet or a solely TMR diet. A total of 4 clusters were identified. Clusters 1 and 3 were mainly associated with samples primarily collected from cows with access to pasture of varying quality and TMR, cluster 2 was formed by samples from cows receiving only TMR, and cluster 4 was a mix of samples from cows receiving high-quality pasture and TMR or TMR only. Interestingly, we found little difference in the relative abundance of methanogens between the community clusters. There was evidence of differences in diversity between pasture associated bacterial communities and those associated with TMR. Cluster 4 had higher diversity and a less robust co-occurrence network based on Spearman correlations than communities representing TMR only or lower-quality pasture samples. These findings indicate that varied bacterial communities are correlated with the metabolic characteristics of different diets. The overall good pasture and TMR quality in this study, combined with the organic allowance for feeding high levels of TMR even during the grazing season, might have contributed to the lack of differences in the fecal archaeal community from samples associated with a mixed pasture and TMR diet, and a TMR only diet. Mitigation strategies to decrease methane emissions such as increasing concentrate to forage ratio, decreasing pasture maturity and adopting grazing systems targeting high quality pasture have been shown to be efficient for pasture-based systems. However, the allowance for organic dairy producers to provide up to an average of 70% of a ruminant's dry matter demand from dry matter fed (e.g., TMR), suggests that reducing enteric methane emissions may require the development of novel dietary strategies independent of pasture management.

Effects of a farm-specific fecal microbial transplant (FMT) product on clinical outcomes and fecal microbiome composition in preweaned dairy calves.

Gastrointestinal disease (GI) is the most common illness in pre-weaned dairy calves. Therefore, effective strategies to manipulate the microbiome of dairy calves under commercial dairy operations are of great importance to improve animal health and reduce antimicrobial usage. The objective of this study was to develop a farm-specific FMT product and to investigate its effects on clinical outcomes and fecal microbial composition of dairy calves. The FMT product was derived from feces from healthy donors (5-24 days of age) raised in the same calf ranch facility as the FMT recipients. Healthy and diarrheic calves were randomly enrolled to a control (n = 115) or FMT (n = 112) treatment group (~36 g of processed fecal matter once daily for 3 days). Fecal samples were collected at enrollment and again 9 days later after the first FMT dose. Although the FMT product was rich in organisms typically known for their beneficial probiotic properties, the FMT therapy did not prevent or ameliorate GI disease in dairy calves. In fact, calves that received FMT were less likely to recover from GI disease, and more likely to die due to GI disease complications. Fecal microbial community analysis revealed an increase in the alpha-diversity in FMT calves; however, no major differences across treatment groups were observed in the beta-diversity analysis. Calves that received FMT had higher relative abundance of an uncultured organism of the genus Lactobacillus and Lactobacillus reuteri on day 10. Moreover, FMT calves had lower relative abundance of Clostridium nexile and Bacteroides vulgatus on day 10. Our results indicate the need to have an established protocol when developing FMT products, based on rigorous inclusion and exclusion criteria for the selection of FMT donors free of potential pathogens, no history of disease or antibiotic treatment.

Fecal microbiome profiles of neonatal dairy calves with varying severities of gastrointestinal disease.

Gastrointestinal disease (GI) is the most common illness in pre-weaned dairy calves. Studies have associated the fecal microbiome composition with health status, but it remains unclear how the microbiome changes across different levels of GI disease and breeds. Our objective was to associate the clinical symptoms of GI disease with the fecal microbiome. Fecal samples were collected from calves (n = 167) of different breeds (Holstein, Jersey, Jersey-cross and beef-cross) from 4-21 d of age. Daily clinical evaluations assessed health status. Calves with loose or watery feces were diagnosed with diarrhea and classified as bright-sick (BS) or depressed-sick (DS) according to behavior. Calves with normal or semiformed feces and no clinical illness were classified as healthy (H). One hundred and three fecal samples were obtained from consistently healthy calves and 64 samples were from calves with diarrhea (n = 39 BS; n = 25 DS). The V3-V4 region of 16S rRNA gene was sequenced and analyzed. Differences were identified by a linear-mixed effects model with a negative binomial error. DS and Jersey calves had a higher relative abundance of Streptococcus gallolyticus relative to H Holstein calves. In addition, DS calves had a lower relative abundance of Bifidobacterium longum and an enrichment of Escherichia coli. Species of the genus Lactobacillus, such as an unclassified Lactobacillus, Lactobacillus reuteri, and Lactobacillus salivarius were enriched in calves with GI disease. Moreover, we created a model to predict GI disease based on the fecal microbiome composition. The presence of Eggerthella lenta, Bifidobacterium longum, and Collinsella aerofaciens were associated with a healthy clinical outcome. Although lactobacilli are often associated with beneficial probiotic properties, the presence of E. coli and Lactobacillus species had the highest coefficients positively associated with GI disease prediction. Our results indicate that there are differences in the fecal microbiome of calves associated with GI disease severity and breed specificities.

Dairy veterinarians' perceptions and experts' opinions regarding implementation of antimicrobial stewardship on dairy farms in the western United States.

OBJECTIVE: To explore veterinarians' perceptions and veterinary experts' opinions regarding antimicrobial stewardship (AMS) on dairy farms in the western United States. SAMPLE: 20 dairy veterinarians and 9 AMS experts. PROCEDURES: 3 focus group discussions involving 20 dairy veterinarians from California, Idaho, and Washington and an expert opinion study involving 9 North American AMS experts were conducted. During focus group discussions, participants were asked open-ended questions regarding implementation of AMS programs on dairy farms. Discussions were recorded and transcribed for thematic analysis. An asynchronous nominal group process was used for the expert opinion study. Participants were asked to complete a series of 3 online surveys consisting of open-ended questions. Expert opinion data underwent thematic analysis and were compared with results obtained from focus group discussions. RESULTS: Veterinarian-perceived barriers to implementation of AMS on dairy farms included variable relationships with clients and farm employees, ensuring AMS provided value to the farm, and uncertainty about regulations for monitoring on-farm antimicrobial use (AMU). Veterinarians were willing to accept additional responsibility for AMU provided that protocols were adopted to ensure them more complete control of on-farm AMU and they were compensated. The AMS experts indicated that effective implementation of AMS on dairy farms requires producer buy-in and tools to facilitate treatment protocol development and monitoring. CONCLUSIONS AND CLINICAL RELEVANCE: Additional veterinary oversight of AMU on dairy farms will require engagement by both veterinarians and producers and practical value-added methods for AMS. Continuing education programs should address treatment protocol development, AMU monitoring strategies, and employee training.

Parenteral Antimicrobial Treatment Diminishes Fecal Bifidobacterium Quantity but Has No Impact on Health in Neonatal Dairy Calves: Data From a Field Trial.

There is evidence that neonatal calves are over treated with antimicrobials that may disrupt colonization of their gastrointestinal tract (GIT) microbiota. The study objectives were to assess the decision-making process of antimicrobial use on a commercial dairy and impacts of parenteral antibiotics on dairy calves' GIT Bifidobacterium and calf health. Unhealthy pre-weaned dairy calves were enrolled based on farm personnel identification with age-matched healthy calves. Half the calves in each group were treated with a 3-day course of IM ampicillin and half were given supportive therapy as needed. Health scores (appetite, fecal consistency, attitude, and temperature) were recorded twice daily throughout the study. Because of inconsistency in employee health decisions, the 121 enrolled calves were reassessed using objective clinical observations plus fecal dry matter and placed into 1 of 3 health categories: healthy, uncomplicated diarrhea (bright attitude and good appetite but with diarrhea), and sick. Accounting for treatment group allocation, this resulted in six post-enrollment health and treatment categories. Calves were followed daily for 14 days post-enrollment and fecal samples collected at 6 time points and Bifidobacterium was quantified from these samples using quantitative PCR. The objective criteria for disease definition reclassified many "unhealthy" calves as uncomplicated diarrhea. Including all calves, on average, the quantity of Bifidobacterium decreased from the day of enrollment (median 8 days of age) across time to 14 days post-enrollment. Calves given an antibiotic the day of enrollment had a greater decrease in Bifidobacterium 4 and 9 days later relative to enrollment Bifidobacterium compared to untreated calves. At enrollment, sick calves and those categorized as uncomplicated diarrhea were more likely to have low Bifidobacterium counts and less likely to be categorized as healthy following antimicrobial treatment. Our results indicate that relying on farm personnel to identify morbidity may lead to some clinical misclassification. There was no indication that antimicrobials affected subsequent health outcomes, but antimicrobials did impact Bifidobacterium dynamics. These results highlight the importance and difficulty in assigning appropriate illness classification on farms and point to a need to develop better point of care diagnostics that improve calf husbandry and stewardship of antimicrobials.

Geographic, farm, and animal factors associated with multiple antimicrobial resistance in fecal Escherichia coli isolates from cattle in the western United States.

OBJECTIVE: To describe geographic, farm-type, and animal-type factors associated with multiple antimicrobial resistance (MAR) in fecal Escherichia coli isolates from cattle. DESIGN: Cross-sectional field study. SAMPLE POPULATION: 1,736 fecal samples from cattle on 38 farms in California, Oregon, and Washington. PROCEDURES: Fecal samples were collected from preweaned calves (2 to 4 weeks old) and cows that recently calved on dairy and beef cow-calf farms, preweaned calves on calf ranches, and 1-year-old steers on feedlots. One fecal E coli isolate per sample was isolated, and antimicrobial susceptibility was tested. Escherichia coli isolates were initially clustered by antimicrobial resistance patterns and categorized by number of antimicrobial resistances. A generalized estimating equations cumulative logistic regression model was used to identify factors associated with an increase in MAR in fecal E coli isolates from cattle. RESULTS: MAR was higher in E coli isolates from cattle in California, compared with those from cattle in Washington or Oregon. Multiple antimicrobial resistance was highest in E coli isolates from calves on calf ranches and progressively lower in isolates from feedlot steers, dairy cattle, and beef cattle. Multiple antimicrobial resistance was higher in E coli isolates from calves than from adult cattle, in E coli isolates from cattle of conventional farms than of organic farms, and in isolates from beef cattle in intensive dairy farm regions than from beef cattle distant from dairy farm regions. CONCLUSIONS AND CLINICAL RELEVANCE: MAR in fecal E coli isolates from cattle was influenced by factors not directly associated with the use of antimicrobials, including geographic region, animal age, and purpose (beef vs dairy).

A Fixed Cohort Field Study of Gene Expression in Circulating Leukocytes From Dairy Cows With and Without Mastitis.

Specifically designed gene expression studies can be used to prioritize candidate genes and identify novel biomarkers affecting resilience against mastitis and other diseases in dairy cattle. The primary goal of this study was to assess whether specific peripheral leukocyte genes expressed differentially in a previous study of dairy cattle with postpartum disease, also would be expressed differentially in peripheral leukocytes from a diverse set of different dairy cattle with moderate to severe clinical mastitis. Four genes were selected for this study due to their differential expression in a previous transcriptomic analysis of circulating leukocytes from dairy cows with and without evidence of early postpartum disease. An additional 15 genes were included based on their cellular, immunologic, and inflammatory functions associated with resistance and tolerance to mastitis. This fixed cohort study was conducted on a conventional dairy in Washington state. Cows >50 days in milk (DIM) with mastitis (n = 12) were enrolled along with healthy cows (n = 8) selected to match the DIM and lactation numbers of mastitic cows. Blood was collected for a complete blood count (CBC), serum biochemistry, leukocyte isolation, and RNA extraction on the day of enrollment and twice more at 6 to 8-days intervals. Latent class analysis was performed to discriminate healthy vs. mastitic cows and to describe disease resolution. RNA samples were processed by the Primate Diagnostic Services Laboratory (University of Washington, Seattle, WA). Gene expression analysis was performed using the Nanostring System (Nanostring Technologies, Seattle, Washington, USA). Of the four genes (C5AR1, CATHL6, LCN2, and PGLYRP1) with evidence of upregulation in cows with mastitis, three of those genes (CATHL6, LCN2, and PGLYRP1) were investigated due to their previously identified association with postpartum disease. These genes are responsible for immunomodulatory molecules that selectively enhance or alter host innate immune defense mechanisms and modulate pathogen-induced inflammatory responses. Although further research is warranted to explain their functional mechanisms and bioactivity in cattle, our findings suggest that these conserved elements of innate immunity have the potential to bridge disease states and target tissues in diverse dairy populations.

Role of ceftiofur in selection and dissemination of blaCMY-2-mediated cephalosporin resistance in Salmonella enterica and commensal Escherichia coli isolates from cattle.

Third-generation cephalosporin resistance of Salmonella and commensal Escherichia coli isolates from cattle in the United States is predominantly conferred by the cephamycinase CMY-2, which inactivates beta-lactam antimicrobial drugs used to treat a wide variety of infections, including pediatric salmonellosis. The emergence and dissemination of bla(CMY-2)(-)-bearing plasmids followed and may in part be the result of selection pressure imposed by the widespread utilization of ceftiofur, a third-generation veterinary cephalosporin. This study assessed the potential effects of ceftiofur on bla(CMY-2) transfer and dissemination by (i) an in vivo experimental study in which calves were inoculated with competent bla(CMY-2)-bearing plasmid donors and susceptible recipients and then subjected to ceftiofur selection and (ii) an observational study to determine whether ceftiofur use in dairy herds is associated with the occurrence and frequency of cephalosporin resistance in Salmonella and commensal E. coli. The first study revealed bla(CMY-2) plasmid transfer in both ceftiofur-treated and untreated calves but detected no enhancement of plasmid transfer associated with ceftiofur treatment. The second study detected no association (P = 0.22) between ceftiofur use and either the occurrence of ceftiofur-resistant salmonellosis or the frequency of cephalosporin resistance in commensal E. coli. However, herds with a history of salmonellosis (including both ceftiofur-resistant and ceftiofur-susceptible Salmonella isolates) used more ceftiofur than herds with no history of salmonellosis (P = 0.03) These findings fail to support a major role for ceftiofur use in the maintenance and dissemination of bla(CMY-2)-bearing plasmid mediated cephalosporin resistance in commensal E. coli and in pathogenic Salmonella in these dairy cattle populations.

Dairy farm soil presents distinct microbiota and varied prevalence of antibiotic resistance across housing areas.

Dairy cattle of different ages experience different living conditions and varied frequency of antibiotic administration that likely influence the distribution of microbiome and resistome in ways that reflect different risks of microbial transmission. To assess the degree of variance in these distributions, fecal and soil samples were collected from six distinct housing areas on commercial dairy farms (n = 7) in Washington State. 16S rRNA gene sequencing indicated that the microbiota differed between different on-farm locations in feces and soil, and in both cases, the microbiota of dairy calves was often distinct from others (P < 0.05). Thirty-two specific antibiotic resistance genes (ARGs) were widely distributed on dairies, of which several clinically relevant ARGs (including cfr, cfrB, and optrA) were identified for the first time at U.S. dairies. Overall, ARGs were observed more frequently in feces and soil from dairy calves and heifers than from hospital, fresh, lactation and dry pens. Droplet-digital PCR demonstrated that the absolute abundance of floR varied greatly across housing areas and this gene was enriched the most in calves and heifers. Furthermore, in an extended analysis with 14 dairies, environmental soils in calf pens had the most antibiotic-resistant Escherichia coli followed by heifer and hospital pens. All soil E. coli isolates (n = 1,905) are resistant to at least 4 different antibiotics, and the PFGE analysis indicated that florfenicol-resistant E. coli is probably shared across geographically-separated farms. This study identified a discrete but predictable distribution of antibiotic resistance genes and organisms, which is important for designing mitigation for higher risk areas on dairy farms.

Dissemination of antimicrobial resistant strains of Campylobacter coli and Campylobacter jejuni among cattle in Washington State and California.

The purpose of this study was to investigate the genetic similarity of Campylobacter jejuni and Campylobacter coli with similar antimicrobial resistance phenotypes, isolated from cattle on different farms and at different times, in order to evaluate the possible existence of disseminated antimicrobial resistant clones. PFGE after SmaI and KpnI restriction identified 23 and 16 distinct PFGE patterns among 29 C. jejuni and 66 C. coli isolates, respectively. In C. coli, 51 (77%) of the resistant isolates demonstrated one of the four indistinguishable PFGE patterns, whereas only 24% doxycycline resistant C. jejuni shared one of the two indistinguishable PFGE patterns. The genetic mechanisms of resistance were homogeneous within and between these clonal types. Genetically indistinguishable (clonal) groups of C. coli accounted for most Campylobacter sp. with multiple antimicrobial resistance observed in this study, consistent with a role for clonal dissemination in the epidemiology of resistance in this species.

Addressing educational challenges in veterinary medicine through the use of distance education.

The veterinary profession is currently facing many educational challenges, including an insufficient capacity to train and educate veterinarians for the multiple disciplines within the profession, a shortage of veterinarians in private and public practice, a shortage of faculty, a lack of human and professional diversity, and a rising cost of education resulting in extreme student debt loads. As a methodology for teaching, distance education (DE) has the potential to address many of these issues. By its very nature, DE can increase the capacity of current facilities and faculty. In addition, DE can allow students to acquire the necessary knowledge at less cost. This article describes a model for incorporating DE in the form of interactive Web-based courses, in conjunction with short, intensive residential programs, for the lecture portions of courses taught in the pre-veterinary, veterinary, and post-veterinary educational periods. In this model, the Web-based courses are used to convey the necessary core knowledge required at each step of the educational process. The residential portions are then used to apply the knowledge in such a way as to combine clinical applications with research in basic and applied sciences. Distance education can provide increased flexibility, high-quality educational experiences, and a less costly alternative for students while maximizing the reach of current faculty efforts and the capacity of existing physical structures.

Stakeholder position paper: dairy producer.

Bacterial antimicrobial resistance is a problem common to both animal and public health. An important public policy issue is to develop and implement prudent use practices where antimicrobials are used. As policy develops, there are questions regarding the use of antimicrobials in animal agriculture and whether these uses constitute prudent use. A series of papers assessing the risk to the public health from agricultural use of antimicrobials have consistently concluded that risk estimation is hampered by the lack of data that describe the amount, types, and uses of antimicrobials in animal agriculture. The absence of information has spurred efforts to develop a framework to collect these data. However, the reasons and benefit of collecting these data should be carefully defined. The dairy industry, contrasted to other major animal commodities, is not focused on meat production but on milk production. Milk production is constrained by disease and antimicrobial treatment is a common management tool, but unlike many other animal agricultural systems where the value and safety of the product is measured in the future; the value of milk is zero when an antimicrobial is used in a lactating cow and milk must be discarded because of residues. While there are exceptions, e.g. non-lactating cow therapy, this difference results in antimicrobials being used sporadically and directed at therapy rather than prophylactic uses. In the dairy industry, antimicrobial use data and its consequences may exist in sufficient detail or could be estimated from existing datasets without the expense of additional surveys. Finally, the main food product milk is mainly pasteurized and all shipments of milk from the farm to the processing plant are tested for the presence of antimicrobials. This makes the likelihood of farm-origin antimicrobials or bacteria appearing in finished product very low. This suggests that the use and quantity of antimicrobials in the dairy system has little impact on public health. This does not imply that the dairy industry does not have a significant role in developing guidelines for appropriate and careful application of antimicrobials, but the effort and cost to collect additional data should be used to fund efforts that improve our diagnostic and managerial skills. These data would change the use of antimicrobials by decreasing the rates of disease and ultimately decreasing prophylactic, metaphylactic and therapeutic use of antimicrobial. These studies and outcome are as important to the dairy industry as to public health.

Stakeholder position paper: epidemiological perspectives on antibiotic use in animals.

Epidemiologists studying antimicrobial resistance are often interested in analyzing the association between antimicrobial resistance and antimicrobial use in animals, and on the impact of antimicrobial use in animals on the occurrence of resistance in bacteria affecting human populations. Given the various potential antimicrobial use data sources, it seems likely there will be some variability in the utility of the data for interpreting trends in antimicrobial resistance and investigating the relationship between antimicrobial use in animals and antimicrobial resistance in bacteria affecting human health. From an epidemiologic perspective, the major issues related to incorporation of antimicrobial use data into antimicrobial resistance monitoring programs are the further development of epidemiologic methods for collecting, quantifying, analyzing and interpreting use data; an open and realistic consideration of the limitations of the data; developing an understanding of scaling, temporal and spatial heterogeneity issues; and the interpretative problems of ecologic and atomistic fallacy. Given the many potential biases in antimicrobial use data, attempts to relate levels of antimicrobial use to levels of antimicrobial resistance should be done with caution until the data are better understood and the aforementioned issues have been addressed.

Prevalence and antimicrobial resistance patterns of Salmonella enterica in preweaned calves from dairies and calf ranches.

OBJECTIVE: To evaluate serovar and antimicrobial resistance patterns of Salmonella enterica isolated from preweaned calves and identify management risk factors associated with fecal shedding of S enterica. SAMPLE POPULATION: Cohorts of 10 to 15 preweaned calves (1 to 84 days of age) on 26 dairies and 7 calf ranches and cross-sectional samples of preweaned calves on smaller farms. PROCEDURES: Calves were evaluated every 2 weeks during a 6-week period. Salmonella isolates obtained from rectal fecal swabs underwent antimicrobial susceptibility testing against 12 antimicrobials. Cluster analysis enabled description of antimicrobial susceptibility patterns. Calf, cohort, and farm risk factors associated with both the prevalence of S enterica and multiple-antimicrobial-resistant S enterica in preweaned calves were identified with repeated-measure logistic regression models. RESULTS: Salmonella enterica was detected on > 50% of farms and in 7.5% of 3,686 fecal samples. Many isolates (33%) were resistant to multiple antimicrobials. Shedding of Salmonella spp was negatively associated with increasing calf age, herds being closed to incoming cattle, and antimicrobial supplementation of milk replacer; prophylactic antimicrobial treatment in day-old calves increased shedding. No association between farm management and presence of multiple-antimicrobial-resistant S enterica or between calving management and presence of S enterica in calves < or = 1 week old was detected. CONCLUSIONS AND CLINICAL RELEVANCE: In preweaned calves, the most important factors associated with decreased likelihood of fecal shedding of S enterica were the use of antimicrobial-supplemented milk replacer and maintenance of a closed herd. Infection with multiple-antimicrobial-resistant S enterica was not associated with antimicrobial administration.