Diagnostic and commensal Staphylococcus pseudintermedius genomes reveal niche adaptation through parallel selection of defense mechanisms.

Staphylococcus pseudintermedius is historically understood as a prevalent commensal and pathogen of dogs, though modern clinical diagnostics reveal an expanded host-range that includes humans. It remains unclear whether differentiation across S. pseudintermedius populations is driven primarily by niche-type or host-species. We sequenced 501 diagnostic and commensal isolates from a hospital, veterinary diagnostic laboratory, and within households in the American Midwest, and performed a comparative genomics investigation contrasting human diagnostic, animal diagnostic, human colonizing, pet colonizing, and household-surface S. pseudintermedius isolates. Though indistinguishable by core and accessory gene architecture, diagnostic isolates harbor more encoded and phenotypic resistance, whereas colonizing and surface isolates harbor similar CRISPR defense systems likely reflective of common household phage exposures. Furthermore, household isolates that persist through anti-staphylococcal decolonization report elevated rates of base-changing mutations in - and parallel evolution of - defense genes, as well as reductions in oxacillin and trimethoprim-sulfamethoxazole susceptibility. Together we report parallel niche-specific bolstering of S. pseudintermedius defense mechanisms through gene acquisition or mutation.

Revision of fluoroquinolone breakpoints used for interpretation of antimicrobial susceptibility testing of canine bacterial isolates.

The fluoroquinolone antimicrobial agents, enrofloxacin and marbofloxacin, were US Food and Drug Administration (FDA) approved in the United States for use in dogs in 1988 and 1999, respectively. There have been many advances since then concerning the pharmacokinetic-pharmacodynamic (PK-PD) evaluation of fluoroquinolones, and there are data available on the susceptibility of targeted pathogens since the original approval. Using this information, the Clinical and Laboratory Standards Institute (CLSI) Veterinary Antimicrobial Susceptibility Testing Subcommittee (VAST) revised its antimicrobial susceptibility testing breakpoints. The previous breakpoints (used in older editions of CLSI standards) for enrofloxacin in dogs were susceptible (S), ≤ 0.5 µg/mL, intermediate (I) 1-2 µg/mL, and resistant (R) ≥ 4 µg/mL. The new breakpoints are S ≤ 0.06 µg/mL for a dose of 5 mg/kg, 0.12 µg/mL for a dose of 10 mg/kg, 0.25 µg/mL for a high dose of 20 mg/kg, and R ≥ 0.5 µg/mL. The breakpoints of 0.12 and 0.25 µg/mL represent a new susceptible-dose dependent (SDD) category. For marbofloxacin, previous breakpoints were S, ≤ 1 µg/mL, I 2 µg/mL, and R ≥ 4 µg/mL. The new breakpoints are S ≤ 0.12 µg/mL for a dose of 2.8 mg/kg, 0.25 µg/mL for a dose of 5.5 mg/kg (SDD), and R ≥ 0.5 µg/mL. The new breakpoints will be published in the next edition of CLSI-Vet01(S) and deviate considerably from the prior breakpoints. Laboratories are encouraged to revise their testing standards. These changes will likely reduce the unnecessary use of these fluoroquinolones in dogs.

Benefits and challenges of creating veterinary antibiograms for empiric antimicrobial selection in support of antimicrobial stewardship and advancement of one-health goals.

Antibiograms are important tools for antimicrobial stewardship that are often underutilized in veterinary medicine. Antibiograms summarize cumulative antimicrobial susceptibility testing (AST) data for specific pathogens over a defined time period; in veterinary medicine, they are often stratified by host species and site of infection. They can aid practitioners with empiric therapy choices and assessment of antimicrobial resistance trends within a population in support of one-health goals for antimicrobial stewardship. For optimal application, consideration must be given to the number of isolates used, the timeframe of sample collection, laboratory analytical methodology, and the patient population contributing to the data (eg, treatment history, geographic region, and production type). There are several limitations to veterinary antibiograms, including a lack of breakpoint availability for bacterial species, a lack of standardization of laboratory methodology and technology for culture and AST, and a lack of funding to staff veterinary diagnostic laboratories at a level that supports antibiogram development and education. It is vital that veterinarians who use antibiograms understand how to apply them in practice and receive relevant information pertaining to the data to utilize the most appropriate antibiogram for their patients. This paper explores the benefits and challenges of developing and using veterinary antibiograms and proposes strategies to enhance their applicability and accuracy. Further detail regarding the application of veterinary antibiograms by privately practicing clinicians is addressed in the companion Currents in One Health article by Lorenz et al (JAVMA, September 2023).

Antimicrobial use in 20 U.S. beef feedyards: 2018-2019.

The objective of this study was to report antimicrobial use in a convenience sample of U.S. beef feedyards for the years 2018 and 2019. In addition to antimicrobial use metrics, also reported are the indications for antimicrobial use and outcomes related to these indications. Antimicrobial use is characterized at the study and feedyard levels for a total of 1,141,846 head of cattle in 20 U.S. feedyards. Antimicrobial use is reported as milligrams of active antimicrobial ingredient per kilogram of liveweight sold (mg/kg-LW) and regimens of antimicrobials per animal year (Reg/AY). Regimens are described by antimicrobial class within use category as characterized by mg of active antimicrobial product per regimen (mg/Reg) and calendar days of administration per regimen (CDoA/Reg). A total of 1,128,515 regimens of medically important antimicrobials were captured from records. The number of regimens/100 head-in (Reg/100 head-in) are described in a subset of 10 feedyards with adequate data granularity to directly determine indications for antimicrobial administration. For the indications of bovine respiratory disease (BRD), Lameness (Lame), Liver Abscess Control (LAC), and Other (e.g., central nervous system disease, cellulitis) the Reg/100 head-in study-level values are 37.1, 0.8, 98.4, and 0.7, respectively, for 2018, with similar values for 2019. The regimens for BRD are further categorized in these 10 feedyards by the use categories in-feed, control of BRD, and individual animal therapy, yielding study level values of 4.6, 19.6, and 12.9 Reg/100 head-in, respectively, for 2018, with similar values for 2019. Outcomes of therapy for individual animal treatment of BRD, Lame, and Other are reported as treatment success, retreatment, or mortality by 30 days after the initial therapy of an animal for a disease. Treatment success rates (no treatment or mortality in the next 30 days) for 2018 in the 10 feedyards with sufficient data granularity are 76.5, 86.5, and 83.0% for BRD, Lame, and Other, respectively. The comparison of these results with other reports of antimicrobial use in North American feedyards highlights how differing approaches in calculating metric values may result in substantially different conclusions regarding antimicrobial use, especially in relation to long-duration uses.

Application and Interpretation of Antimicrobial Susceptibility Testing.

Antimicrobial susceptibility testing (AST) is an important component of antimicrobial stewardship. This article discusses how AST methods and breakpoints are developed, describes when AST may or may not be useful in clinical practice, and discusses how to interpret AST results from bacterial isolates from cattle, sheep, and goats. Discussion of when AST is not appropriate or when veterinarians should have low confidence in AST results is also included. Applicability of genomic testing for antimicrobial susceptibility is briefly addressed.

Nasopharyngeal Bacterial Prevalence and Microbial Diversity at First Treatment for Bovine Respiratory Disease (BRD) and Its Associations with Health and Mortality Outcomes in Feedyard Cattle.

Bovine respiratory disease (BRD) is an economically important disease in feedyards influencing both animal welfare and antimicrobial utilization. Major pathogens associated with BRD have been identified in previous research, but little information is available on the relationship between nasopharyngeal microbiota and health outcomes. The objective of this study was to identify potential associations between nasopharyngeal microbiota and antimicrobial resistance patterns of clinical cases that lived or died compared to non-diseased controls. Enrolled animals were subdivided based on clinical disease status and case outcome (subsequent mortality). Deep nasopharyngeal swabs were collected on enrolled animals and submitted for bacterial isolation, antimicrobial susceptibility determination, and metagenomics analysis. Enrolled cattle were represented in three groups: animals at first treatment for BRD that subsequently died (BRDM, n = 9), animals at first treatment for BRD that subsequently lived (BRDL, n = 15), and animals that were never treated for BRD during the feeding phase (CONT, n = 11). Antimicrobial resistance patterns for Pasteurella multocida illustrated cattle in each outcome category had isolates that were pan-susceptible or only showed resistance to oxytetracycline. The relative abundance of species and genera illustrated few differences among the three outcomes. Higher alpha diversity was identified in BRDL compared to CONT at the species level, and both BRDL and BRDM showed increased alpha diversity compared to CONT at the general level. Overall, this work illustrated nasopharyngeal microbiota showed relatively few differences among BRD cases that lived or died compared to animals without BRD.

Improving farm-level antimicrobial stewardship benchmarks by reporting antimicrobial use within the context of both the magnitude of disease pressure and the outcome of therapy.

This manuscript explores a method of benchmarking antimicrobial use within the context of farm level therapeutic incidence (a proxy for disease incidence), and the outcome of that therapy. This is reported both within the same farm over time (2016-2019), as well as evaluated across participating farms. Reporting antimicrobial use in this format addresses multiple primary questions necessary for evaluating on farm antimicrobial stewardship: How much disease is recorded? How much antimicrobial use is recorded? How often are antimicrobials included in therapy for each disease? What is the outcome of therapy? The three primary metrics reported are: therapeutic events per 100 cow years (TE/100CY), antimicrobial regimens per 100 cow years (REG/100CY), and the percent therapeutic success (% Success). Success was defined as: the cow remained in the herd and had no further TE recorded within 30 days of the end of the TE being evaluated. These measures identify opportunities for change on an individual farm, such as improvement in disease prevention, or a change in choices about when to include an antimicrobial in the treatment protocol. Therapeutic outcomes provide additional context, in some instances demonstrating differences in recording practices and case definitions, while in other cases serving to safeguard animal welfare as efforts are made to decrease antimicrobial use in the future. Although developed for farm level reporting, the metrics may also be more broadly summarized to meet future reporting requirements for marketing chain or national level antimicrobial use reports. The process outlined here serves as a prototype to be considered when developing antimicrobial use reporting systems where farm level antimicrobial stewardship is the primary objective.

The association of multiple metrics for evaluating antimicrobial use in U.S. beef feedyards.

In order to accurately portray antimicrobial use in food animals, the need for standardized metrics, and an understanding of the characteristics of different metrics, has long been recognized. Fourteen U.S. feedyards were used to evaluate the effects of using centralized constants such as defined daily dose (DDD) and defined course dose (DCD) applied to the weight of medically important antimicrobials by class (mg) as opposed to using electronic individual animal treatment records and lot level in-feed antimicrobial records obtained from the same population. Three numerators were calculated directly from recorded data for each drug product: the number of antimicrobial regimens associated with indication (Reg), milligrams of drug administered per regimen (mg), and calendar days of administration for each regimen (CDoA). There were four use indications to which numerators were assigned: liver abscess control (LAC), bovine respiratory disease (BRD), lameness (lame), or all other indications combined (other). Three denominators were also calculated directly from the data, these being the number of days animals were present (head days), number of cattle received (head in), and kilograms of live weight sold (kg-LW). Numerators and denominators were calculated at the lot level. The use of DDD or DCD was explored to determine how their use would affect interpretation of comparisons between lots or feedyards. At the lot level across both study years, the lot estimate of nDDD differed from the CDoA value by >25% in 49.2% of the lots. The number of Defined Course Doses (nDCD) was then compared to the number of Regimens (Reg). Comparing nDCD to Reg at the lot level across both study years, the lot estimate of nDCD differed from the Reg value by >25% in 46.4% of lots. Both year and metric were also shown to affect numerical feedyard ranking by antimicrobial use according to seven different metrics. The analysis reported here adds to the body of literature reporting substantial effects of metric choice on the conclusions drawn from comparing antimicrobial use across multiple production sites.

Using telehealth clinical case vignettes to enhance clinical confidence and competence in veterinary students.

Veterinarians contribute substantially to the health of their patients and enhance the communities in which they live. Delivery of veterinary curricula continues to evolve to ensure that veterinary graduates are prepared to meet their professional obligations on Day One of their careers. In this study, veterinary practitioners were recruited to deliver telehealth case rounds to veterinary students at Kansas State University and Texas A&M University. Case discussions were hosted virtually once per month in the 2020-2021 and 2021-2022 academic years for a total of 16 sessions. Each presenting practitioner was instructed to develop a brief presentation for a case routinely seen in their practice, and to discuss important clinical decision points in diagnosis, treatment and management. Cases could also highlight important ethical or communication issues encountered in veterinary medicine. The overall goals of this project were to increase the quantity and diversity of clinical cases to which veterinary students were exposed during their professional training and to evaluate the feasibility and acceptability of telehealth technology as a teaching strategy. Student participants were surveyed to determine the effectiveness of telehealth sessions in increasing overall confidence and competence in case management, and veterinary presenters were surveyed to determine motivations for participating in the project and perceived value of the telehealth sessions. More than 95% of students indicated that participation in telehealth sessions increased their clinical confidence and competence. Presenting practitioners unanimously indicated that they would participate in similar instruction in the future. Recommendations are provided to improve the educational experience for future adopters of telehealth teaching sessions.

Synergistic combinations of clarithromycin with doxycycline or minocycline reduce the emergence of antimicrobial resistance in Rhodococcus equi.

BACKGROUND: The alarming increase in rifampin and macrolide resistance among Rhodococcus equi isolates highlights the need to identify alternative therapeutic options that can effectively control rhodococcosis in foals while limiting the further development of drug resistance. OBJECTIVES: To evaluate bacterial killing, antibiotic synergism and mutant prevention concentrations (MPCs) of clarithromycin alone and in combination with doxycycline, minocycline or rifampin against clinical isolates of R equi. STUDY DESIGN: In vitro experiments. METHODS: Bacterial time-kill, fractional inhibitory concentration (checkerboard) and mutant prevention concentration assays were evaluated in four clarithromycin- and rifampin-susceptible (ClaS /RifS ) and two clarithromycin- and rifampin-resistant (ClaR /RifR ) R equi clinical strains. RESULTS: In this study evaluating a limited number of isolates, combinations of clarithromycin with doxycycline or minocycline, but not with rifampin, were generally synergistic in both ClaS /RifS and ClaR /RifR strains as determined by checkerboard testing. In time-kill assays, all antibiotics, both alone and in combination, reduced viable ClaS /RifS R equi by more than 3 log10 at 24 hours compared with control cultures without antibiotics. Combinations of clarithromycin with doxycycline, minocycline or rifampin induced significantly lower MPC values compared with the individual antimicrobials alone for all ClaS /RifS R equi strains, resulting in a narrower mutant selection window (MSW). However, clarithromycin/rifampin combination did not markedly decrease MPCs of the individual antimicrobials in ClaR /RifR R equi isolates, and the observed decrease in MPCs for doxycycline or minocycline did not generally differ when combined with clarithromycin. MAIN LIMITATIONS: The number of analysed R equi isolates was limited. In vitro outcomes require clinical confirmation. CONCLUSIONS: Dual therapy combinations consisting of clarithromycin with doxycycline or minocycline merit consideration as a treatment protocol against R equi in foals due to in vitro synergy. These combination therapies may also minimise the emergence of antimicrobial resistance in cases of rhodococcosis.

Detection of bovine inflammatory cytokines IL-1ß, IL-6, and TNF-α with a multiplex electrochemiluminescent assay platform.

Commercially available bovine-specific assays are limited in number, and multiplex assays for this species are rare. Our objective was to develop a multiplex assay for the bovine inflammatory cytokines IL-1ß, IL-6, and TNF-α using the Meso Scale Discovery U-PLEX platform. "Do-It-Yourself" ELISA kits that contained polyclonal antibodies, both unlabeled and biotinylated, and the specific recombinant bovine cytokine standard, were purchased for each of these three cytokines. The biotinylated antibodies were coupled to linkers that bind to specific locations within each well of the U-PLEX plate. Unique linkers were used for each of the cytokines. The unlabeled antibodies were conjugated with electrochemiluminescent labels to serve as detection antibodies. Each cytokine assay was optimized individually prior to performing an optimization on the multiplex assay containing reagents for all three cytokines. To calculate cytokine concentrations, standard curves were developed using the recombinant cytokines and were run concurrently on each plate. Standard curves for IL-1ß and TNF-α were run at concentrations ranging from 0 to 50,000 pg/mL, and for IL-6 from 0 to 10,000 pg/mL. The average lowest level of detection concentration measured by the standard curves were 5.3 pg/mL, 0.92 pg/mL, and 22.34 pg/mL for IL-1ß, IL-6, and TNF-α respectively, as determined by data from seven plates containing bovine plasma samples from a combination of healthy and diseased cattle. The U-PLEX platform was a viable means to develop custom analyte- and species-specific multiplex assays using privately developed or purchased sets of commercially available reagents.

Cytokine and Haptoglobin Profiles From Shipping Through Sickness and Recovery in Metaphylaxis- or Un-Treated Cattle.

Fifty-six head of cattle, 28 animals with bovine respiratory disease complex (BRDC), and 28 healthy animals that were matched by treatment, sale barn of origin, day, and interactions among these variables, were identified from a population of 180 animals (60 each purchased at three sale barns located in Missouri, Tennessee, and Kentucky) enrolled in a study comparing animals receiving metaphylaxis to saline-treated controls. Cattle were transported to a feedlot in KS and assigned to treatment group. Blood samples were collected at Day 0 (at sale barn), Day 1, Day 9, and Day 28 (at KS feedlot), and transported to the US Meat Animal Research Center in Clay Center, NE where plasma was harvested and stored at -80°C until assayed for the cytokines IFN-γ, IL-1ß, IL-6, and TNF-α, and the acute stress protein haptoglobin (HPT). Our objectives were to determine if cytokine and haptoglobin profiles differed between control and metaphylaxis treatment groups over time, and if profiles differed between animals presenting with BRDC and those that remained healthy. There was no difference between the treated animals and their non-treated counterparts for any of the analytes measured. Sale barn of origin tended to affect TNF-α concentration. Differences for all analytes changed over days, and on specific days was associated with state of origin and treatment. The Treatment by Day by Case interaction was significant for HPT. The analyte most associated with BRDC was HPT on D9, possibly indicating that many of the cattle were not exposed to respiratory pathogens prior to entering the feedlot.

Current and future perspectives on the categorization of antimicrobials used in veterinary medicine.

The emergence of antimicrobial resistance in human and veterinary bacterial pathogens has led to concerns regarding the use of antimicrobials in veterinary medicine. Consequently, regulatory agencies have developed procedures for assessing the risk associated with the use of a specific antimicrobial as part of the drug approval process. Due consideration for the importance (priority categorization) of the antimicrobial to human medicine is part of this risk assessment process. Additionally, nongovernmental organizations have developed antimicrobial categorization schemes to protect the use and effectiveness of these medicines. However, the goals and methods of the various categorization schemes vary, resulting in final categorizations that are different. Although harmonizing these schemes would bring clarity to antimicrobial resistance discussions and policy, it has the disadvantage of not accounting for regional antimicrobial resistance and use, potentially removing effective medicines from clinical use in situations where they are wholly appropriate. Antimicrobials should be classified in a One Health manner, where both physician and veterinarian share the responsibility for antimicrobial use. The purpose of this article is to summarize current antimicrobial categorization schemes using illustrative examples to highlight differences and provide perspectives on the impact of the current schemes and future directions.

Pharmacological considerations of antibiotic failures in bovine respiratory disease cases.

Bovine respiratory disease (BRD) is one of the most common indications for antimicrobial therapy in beef cattle production and research trials demonstrate that antibiotic therapy greatly improves clinical outcome for BRD. These trials also show that BRD treatment success rates are less than 100% and that there are opportunities to optimize antimicrobial prescribing and improve clinical outcomes if the underlying cause(s) of BRD treatment failures can be identified and addressed. As the etiology of BRD in an individual animal is frequently multi-factorial in nature; it is likely that BRD treatment failures also result from complex interactions between the drug, drug administrator, animal host, pathogens, and the environment. This review will focus specifically on the pharmacological aspects, specifically the interactions between the host and the drug and the drug and the drug administrator, of BRD treatment failures and the actions that veterinary practitioners can take to investigate and mitigate therapeutic failures in future cases.

Antimicrobial use in 22 U.S. beef feedyards: 2016-2017.

This study describes antimicrobial use in 22 U.S. beef feedyards by use of two metrics: milligrams of antimicrobial per kilogram of liveweight sold (mg/kg-LW) and antimicrobial regimens per animal year (Reg/AY). The primary objectives were to determine the feasibility of collecting antimicrobial use data at the level of the production system and to identify factors unique to feedyard systems which may confound interpretation of summarized antimicrobial use measures. Records were accessed directly from feedyards or through a data intermediary and then subjected to a process of standardization and quality assurance. Use is reported at the study level with all individual cattle lots from all feedyards combined, as well as at the feedyard level where values were first calculated for each feedyard and then expressed as means, standard deviations and median values. Use at the study level is first reported by antimicrobial class and then by antimicrobial class within the use categories of in-feed, control of bovine respiratory disease (BRD) and individual animal treatment. Due to wide variations of antimicrobial class within use category, feedyard level antimicrobial use is reported only by antimicrobial class. Use values for medically important and not medically important antimicrobial classes are reported separately. Regimens are described for each antimicrobial class within use category by milligrams per regimen and by timeframe from first to last administration. The authors' selection of measures reported here was driven by a desire to minimize necessary assumptions, resulting in transparent reporting of values which can be directly related back to the factors within feedyard systems which may have influenced calculations. Reporting the number of regimens stratified by antimicrobial class within use category and then describing the characteristics of the reported regimens (milligrams per regimen and timeframe) optimize these objectives for reporting.

Comparison of surveys and use records for quantifying medically important antimicrobial use in 18 U.S. Beef Feedyards.

The objective of this study was to evaluate agreement between medically important antimicrobial use metrics derived from in-person surveys of feedyard management as opposed to metrics derived from production unit-level antimicrobial use records. Survey respondents were asked to estimate values which would allow calculation of the metrics of regimens per animal year (Reg/AY) and milligrams of antimicrobial per kilogram of liveweight sold (mg/kg-LW). At the study level, values were calculated by antimicrobial class within the use categories of in-feed use, control of bovine respiratory disease (BRD) and individual animal treatment. At the feedyard level, values were calculated by total overall use and total use within use category. Feedyard level correlation coefficients between survey and record values for total use were 0.76 (p = .0004) and 0.73 (p = .0009) for Reg/AY and mg/kg-LW, respectively. Correlation coefficients for use category within metric ranged from 0.25 (p = .3224) to 0.65 (p = .0061). Comparing feedyard level survey and record values for total Reg/AY and mg/kg-LW using a Wilcoxon signed-rank test resulted in p-values (95% CI) of 0.3247 (-1.06, 0.25) and 0.7019 (-14.49, 10.387), respectively. Evaluation of comparative rankings as total use by metric indicated that for Reg/AY, only two of the five top-ranked feedyards were consistent between the two data sources. The relationship for mg/kg-LW demonstrated the lowest two and highest three values were consistent between sources; however, the ranking similarities appear to markedly decline in the middle ranks. This report demonstrates that survey-based antimicrobial use data may closely reflect summary values determined from records across multiple beef feedyards. However, individual feedyard relationships between their record and survey values vary widely and ranking by survey may lead to different conclusions as to highest and lowest use than ranking by data collected from use records.

Antimicrobial use quantification in adult dairy cows - Part 3 - Use measured by standardized regimens and grams on 29 dairies in the United States.

This study describes antimicrobial use in terms of standardized regimens per cow year (REG/CY) and grams per cow year (GMS/CY) for 29 dairies in the United States during the years 2016 and 2017. To explore potential priorities for antimicrobial stewardship programs, these measures were stratified by both disease syndrome and antimicrobial class. Potential confounders of use measurements are discussed and challenges for measure interpretation are identified. When measured as REG/CY, the results indicate that mastitis is the disease syndrome with the greatest contribution to overall antimicrobial use. However, when GMS/CY is measured, metritis, lameness and unknown disease syndromes are also significant contributors. When use is stratified by antimicrobial class, measures of REG/CY indicate the greatest magnitude of use is the cephalosporin class. However, when measures of GMS/CY are stratified by drug class, use within the penicillin class contributes more than any other single class. These differences highlight the need for a more complete understanding of the relationship between antimicrobial use measures and their relationship to antimicrobial resistance selection pressure.

Antimicrobial use quantification in adult dairy cows - Part 1 - Standardized regimens as a method for describing antimicrobial use.

This study describes a process to acquire and convert farm treatment records into a standardized regimen format. Multiple sources of on-farm data were utilized to convert the original treatment records to standardized regimens, enabling the generation of objective, granular descriptions of antimicrobial use. These standard regimen descriptions allow antimicrobial use data to be qualitatively stratified by active substance, disease syndrome treated, use category, and route of administration. Quantitative distributions are available for the grams of active substance per administration, the grams of active substance per regimen, the number of administrations, number of days of therapy and the time frame between the first and last administration. Granular quantification by this method informs future research, surveillance and policy.

Antimicrobial use quantification in adult dairy cows - Part 2 - Developing a foundation for pharmacoepidemiology by comparing measurement methods.

As stewardship programmes seek to influence therapeutic choices and reduce resistance selection pressure, it is essential to understand the multiple factors which may influence any given antimicrobial use measure. This article compares 9 measures of antimicrobial use pertaining to adult cows in US dairy systems based on treatment records standardized to a single standardized regimen (SReg) format. There are 3 fundamental parameters underlying the nine measures: amount of active substance (AMOUNT), number of SRegs (ANIMALS) and treatment time (TIME). Additionally, three use categories were considered in the analysis: control of mastitis (Dry Cow), treatment of detected mastitis (Clinical Mastitis) and treatment of other diseases (Other Treatment). The assumptions necessary for calculation of each of the nine measures are identified. The influence of measure choice on farm rank was explored at the farm level both within and across fundamental parameters. Across all use categories, when measures of TIME and ANIMALS were compared, correlations between measures were greater than 0.91. However, when measures of AMOUNT were compared to either measures of TIME or ANIMALS, the correlation between measures was more variable across use categories (R = 0.31-0.91). These comparisons demonstrate that the selection of antimicrobial use metric can affect which dairies are considered the highest users of antimicrobials. Measurement selection also influences which use category has the greatest potential for use reduction. This indicates that measure selection can influence which use category is prioritized as a target for reduction of antimicrobial use. Utilization of this information may allow those developing dairy antimicrobial stewardship programmes to better understand the potential effects of measurement selection on driving changes in antimicrobial use.