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Plasma chlortetracycline (CTC) concentration data were subjected to Monte Carlo simulation of area under the concentration curve (AUC) values related to bovine respiratory disease pathogen MIC distributions to evaluate target attainment rates. Crossbred Hereford heifers were randomly assigned into two treatment groups. Treatment group (A) received chlortetracycline (CTC) at a target dose of 22 mg/kg of bodyweight daily for 5 consecutive days (n = 8) and group (B) received CTC at 350 mg/head per day (1.5 ± 0.2 mg/kg based on actual bodyweights) for seven consecutive days (n = 8). Non-compartmental analysis was used to calculate plasma-free drug CTC area under the concentration curves. The mean observed (±SD) free drug AUC values were 4.18 (±1.72) µg × h/mL and 0.30 (±0.06) µg × h/mL for treatment groups A and B, respectively. The probability of target attainment for AUC24/MIC values of 25 and 12.5 was modeled using Monte Carlo simulations. Treatment group A achieved >90% target attainment (AUC24/MIC of 25) at an MIC of 0.06 µg/mL, whereas treatment group B displayed only 12.6% target attainment (AUC24/MIC of 12.5) at the lowest MIC evaluated (0.015 µg/mL). Both in-feed CTC regimens failed to obtain a reasonable target attainment rate in light of expected MIC distributions of potential pathogens.
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BACKGROUND: Mannheimia haemolytica strains isolated from North American cattle have been classified into two genotypes (1 and 2). Although members of both genotypes have been isolated from the upper and lower respiratory tracts of cattle with or without bovine respiratory disease (BRD), genotype 2 strains are much more frequently isolated from diseased lungs than genotype 1 strains. The mechanisms behind the increased association of genotype 2 M. haemolytica with BRD are not fully understood. To address that, and to search for interventions against genotype 2 M. haemolytica, complete, closed chromosome assemblies for 35 genotype 1 and 34 genotype 2 strains were generated and compared. Searches were conducted for the pan genome, core genes shared between the genotypes, and for genes specific to either genotype. Additionally, genes encoding outer membrane proteins (OMPs) specific to genotype 2 M. haemolytica were identified, and the diversity of their protein isoforms was characterized with predominantly unassembled, short-read genomic sequences for up to 1075 additional strains. RESULTS: The pan genome of the 69 sequenced M. haemolytica strains consisted of 3111 genes, of which 1880 comprised a shared core between the genotypes. A core of 112 and 179 genes or gene variants were specific to genotype 1 and 2, respectively. Seven genes encoding predicted OMPs; a peptidase S6, a ligand-gated channel, an autotransporter outer membrane beta-barrel domain-containing protein (AOMB-BD-CP), a porin, and three different trimeric autotransporter adhesins were specific to genotype 2 as their genotype 1 homologs were either pseudogenes, or not detected. The AOMB-BD-CP gene, however, appeared to be truncated across all examined genotype 2 strains and to likely encode dysfunctional protein. Homologous gene sequences from additional M. haemolytica strains confirmed the specificity of the remaining six genotype 2 OMP genes and revealed they encoded low isoform diversity at the population level. CONCLUSION: Genotype 2 M. haemolytica possess genes encoding conserved OMPs not found intact in more commensally prone genotype 1 strains. Some of the genotype 2 specific genes identified in this study are likely to have important biological roles in the pathogenicity of genotype 2 M. haemolytica, which is the primary bacterial cause of BRD.
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Proteínas da Membrana Bacteriana Externa/genética , Doenças dos Bovinos/microbiologia , Mannheimia haemolytica/genética , Infecções Respiratórias/veterinária , Sequenciamento Completo do Genoma/métodos , Animais , Bovinos , Cromossomos Bacterianos/genética , Genótipo , Mannheimia haemolytica/classificação , Mannheimia haemolytica/isolamento & purificação , Mutação , FilogeniaRESUMO
Antibiotics can be administered orally or parenterally in swine production, which may influence antimicrobial resistance (AMR) development in gut bacteria. A total of 40 barrows and 40 gilts were used to determine the effects of tylosin administration route on growth performance and fecal enterococcal AMR. The antibiotic treatments followed Food and Drug Administration label directions and were as follows: (1) no antibiotic (CON), (2) 110 mg tylosin per kg feed for 21 d (IN-FEED), (3) 8.82 mg tylosin per kg body weight through intramuscular injection twice daily for the first 3 d of each week for 3 weeks (IM), and (4) 66 mg tylosin per liter of drinking water (IN-WATER). Antibiotics were administered during d 0 to 21 and all pigs were then fed the CON diet from d 21 to 35. Fecal samples were collected on d 0, 21, and 35. Antimicrobial susceptibility was determined by microbroth dilution method. No evidence of route × sex interaction (p > 0.55) was observed for growth performance. From d 0 to 21, pigs receiving CON and IN-FEED had greater (p < 0.05) average daily gain (ADG) than those receiving IM, with the IN-WATER group showing intermediate ADG. Pigs receiving CON had greater (p < 0.05) gain-to-feed ratio (G:F) than IM and IN-WATER, but were not different from pigs receiving IN-FEED. Overall, enterococcal isolates collected from pigs receiving IN-FEED or IM were more resistant (p < 0.05) to erythromycin and tylosin than CON and IN-WATER groups. Regardless of administration route, the estimated probability of AMR to these two antibiotics was greater on d 21 and 35 than on d 0. In summary, IM tylosin decreased ADG and G:F in finishing pigs, which may be because of a response to the handling during injection administration. Tylosin administration through injection and feed resulted in greater probability of enterococcal AMR to erythromycin and tylosin compared with in-water treatment.
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Antibacterianos/administração & dosagem , Farmacorresistência Bacteriana/efeitos dos fármacos , Enterococcus/efeitos dos fármacos , Doenças dos Suínos/prevenção & controle , Tilosina/administração & dosagem , Ração Animal/análise , Animais , Peso Corporal/efeitos dos fármacos , Dieta/veterinária , Esquema de Medicação , Enterococcus/isolamento & purificação , Eritromicina/administração & dosagem , Fezes/microbiologia , Feminino , Masculino , Suínos , Doenças dos Suínos/microbiologia , DesmameRESUMO
OBJECTIVE: To identify conjunctival bacterial flora in healthy adult and newborn kid goats in the Midwestern United States and to compare vaginal and ocular surface flora in dam-kid pairs. ANIMALS STUDIED: Thirty mixed-breed (crosses between Boer, Kiko, and Syfan Spanish) goats, 20 adult does and 10 newborn kids. PROCEDURES: One eye of 15 adult goats (n = 15 eyes) and 5 kids (n = 5 eyes) was randomly selected. A subset of 5 adults (n = 10 eyes) and 5 kids (n = 10 eyes) underwent bilateral sampling. Each recently kidded dam's vaginal canal (n = 10) was also sampled. Two swabs were collected from each sample site for aerobic bacterial culture and Mycoplasma and Chlamydia spp. PCR. RESULTS: Of the animals with positive cultures, Staphylococcus and Streptococcus were the most common bacterial genera from the conjunctival sac of adult (16/17; 94%) and kid (5/5; 100%) goats; three adults and 5 kids had no growth of bacteria on aerobic culture. Moraxella bovoculi was the most common single bacteria in adults, in 9 eyes (36%) of 8 animals (40%). Staphylococcus equorum was identified in all 5 kids with positive cultures. Mycoplasma sp. DNA was detected in 7 animals. Chlamydia sp. DNA was not detected in any sample. Four of 10 dam-kid pairs had identical bacteria isolated from the dam's vaginal sample and the kid's conjunctival sample. CONCLUSIONS: Staphylococcus and Streptococcus were the most common conjunctival bacterial genera in this goatherd. Moraxella bovoculi was the most common single bacteria isolated from adults, and Staphylococcus equorum was the most common bacteria in kids. Mycoplasma sp. occurred infrequently at the ocular surface of adult and kid goats. A convincing association between dam-kid vaginal-conjunctival samples was not identified.
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Túnica Conjuntiva/microbiologia , Cabras/microbiologia , Animais , Animais Recém-Nascidos/microbiologia , Chlamydia , Feminino , Microbiota , Meio-Oeste dos Estados Unidos , Mycoplasma , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase em Tempo Real , Staphylococcus , Streptococcus , Vagina/microbiologiaRESUMO
BACKGROUND: Mannheimia haemolytica typically resides in cattle as a commensal member of the upper respiratory tract microbiome. However, some strains can invade their lungs and cause respiratory disease and death, including those with multi-drug resistance. A nucleotide polymorphism typing system was developed for M. haemolytica from the genome sequences of 1133 North American isolates, and used to identify genetic differences between isolates from the lungs and upper respiratory tract of cattle with and without clinical signs of respiratory disease. RESULTS: A total of 26,081 nucleotide polymorphisms were characterized after quality control filtering of 48,403 putative polymorphisms. Phylogenetic analyses of nucleotide polymorphism genotypes split M. haemolytica into two major genotypes (1 and 2) that each were further divided into multiple subtypes. Multiple polymorphisms were identified with alleles that tagged genotypes 1 or 2, and their respective subtypes. Only genotype 2 M. haemolytica associated with the lungs of diseased cattle and the sequence of a particular integrative and conjugative element (ICE). Additionally, isolates belonging to one subtype of genotype 2 (2b), had the majority of antibiotic resistance genes detected in this study, which were assorted into seven combinations that ranged from 1 to 12 resistance genes. CONCLUSIONS: Typing of diverse M. haemolytica by nucleotide polymorphism genotypes successfully identified associations with diseased cattle lungs, ICE sequence, and antibiotic resistance genes. Management of cattle by their carriage of M. haemolytica could be an effective intervention strategy to reduce the prevalence of respiratory disease and supplemental needs for antibiotic treatments in North American herds.
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Conjugação Genética , Farmacorresistência Bacteriana , Genoma Bacteriano , Genômica , Mannheimia haemolytica/efeitos dos fármacos , Mannheimia haemolytica/fisiologia , Pneumonia Enzoótica dos Bezerros/microbiologia , Animais , Antibacterianos/farmacologia , Bovinos , Ligação Genética , Genômica/métodos , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Mannheimia haemolytica/classificação , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Groundwater provides a reliable tap to sustain agricultural production, yet persistent aquifer depletion threatens future sustainability. The High Plains Aquifer supplies 30% of the nation's irrigated groundwater, and the Kansas portion supports the congressional district with the highest market value for agriculture in the nation. We project groundwater declines to assess when the study area might run out of water, and comprehensively forecast the impacts of reduced pumping on corn and cattle production. So far, 30% of the groundwater has been pumped and another 39% will be depleted over the next 50 y given existing trends. Recharge supplies 15% of current pumping and would take an average of 500-1,300 y to completely refill a depleted aquifer. Significant declines in the region's pumping rates will occur over the next 15-20 y given current trends, yet irrigated agricultural production might increase through 2040 because of projected increases in water use efficiencies in corn production. Water use reductions of 20% today would cut agricultural production to the levels of 15-20 y ago, the time of peak agricultural production would extend to the 2070s, and production beyond 2070 would significantly exceed that projected without reduced pumping. Scenarios evaluate incremental reductions of current pumping by 20-80%, the latter rate approaching natural recharge. Findings substantiate that saving more water today would result in increased net production due to projected future increases in crop water use efficiencies. Society has an opportunity now to make changes with tremendous implications for future sustainability and livability.
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Agricultura , Água Subterrânea , Irrigação Agrícola/tendências , Agricultura/tendências , Animais , Bovinos , Conservação dos Recursos Naturais/tendências , Previsões , Kansas , Recursos Hídricos , Abastecimento de Água , Zea mays/crescimento & desenvolvimentoRESUMO
Supply chain issues disrupt veterinary care and cause downstream consequences that alter the practice of veterinary medicine. Antimicrobials are just 1 class of pharmaceuticals that have been impacted by supply chain issues over the last couple of years. Since February 2021, 2 sponsors/manufacturers of penicillin products have reported shortages in the active pharmaceutical ingredient. With the release of the 2021 Summary Report on Antimicrobials Sold or Distributed for Use in Food-Producing Animals by the FDA, a key finding was a 19% decrease in penicillin sales and distribution from 2020 to 2021. Herein, we provide our clinicians' professional perspective regarding how drug shortages, specifically that of penicillin, might contribute to misconstrued patterns in antimicrobial use and what can be done by veterinarians and the FDA to minimize the impact of an antimicrobial drug shortage on animal health and well-being.
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Anti-Infecciosos , Drogas Veterinárias , Animais , Antibacterianos/uso terapêutico , Artefatos , Anti-Infecciosos/uso terapêutico , PenicilinasRESUMO
A key component of antimicrobial stewardship is the ability to collect antimicrobial use data and ultimately use this information to ensure that administrations are necessary and effective. National antimicrobial sales data cannot help in this capacity because the data lack context, for example, details concerning target species and disease indication. The objective of this study was to continue the development of a system for collecting flock-level on-farm antimicrobial use data from the U.S. turkey industry and to have it be representative of national turkey production in the U.S. This study utilized a public-private partnership to enable collection and protection of sensitive flock-level data from an extremely large industry while releasing deidentified and aggregated information regarding the details of antimicrobial use on U.S. turkey farms over time. Participation was voluntary. Data were collected for the period 2013 through 2021 and are reported on a calendar year basis. Using production statistics from USDA:NASS as a denominator, the data supplied by participating companies represented approximately 67.3% of turkey production in the U.S. in 2013, approximately 69.1% in 2017, and approximately 71.4% in 2021. The data that were submitted for 2021 are based on approximately 149,000,192 turkeys slaughtered and 4,929,773,506 pounds liveweight produced. Detailed prescription records representing approximately 60-70% of the birds were available for the 2018-2021 dataset. The estimated percentage of turkey poults placed that received hatchery antimicrobials decreased from 96.9% in 2013 to 40.5% in 2021. The use of in-feed antimicrobials was practically eliminated, with in-feed tetracycline being the only medically important antimicrobial used in 2021. Use of in-feed tetracyclines decreased approximately 80% between 2013 and 2021. Water-soluble antimicrobial use declined over the study period. Between 2013 and 2021, water-soluble penicillin use decreased approximately 41% but water-soluble tetracycline use increased approximately 22%. Key diseases that were treated with water-soluble antimicrobials included bacterial poult enteritis and clostridial dermatitis. Efforts to reduce the incidence of these diseases would reduce the need for antimicrobial therapy, thereby enabling continued decreases in antimicrobial use without sacrificing animal welfare. However, this will require an investment in research to find efficacious and cost-effective mitigation strategies.
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Although efforts to improve antimicrobial stewardship should include the collection of antimicrobial use data, most antimicrobial datasets collected at the national level consist of antimicrobial sales data which cannot inform stewardship. These data lack context, such as information regarding target species, disease indication, and regimen specifics like dose, route and duration. Therefore, the goal of this study was to develop a system for collecting data on the use of antimicrobials in the U.S. broiler chicken industry. This study utilized a public-private partnership to enable collection and protection of sensitive data from an extremely large industry while releasing deidentified and aggregated information regarding the details of antimicrobial use on U.S. broiler chicken farms over time. Participation was voluntary. Data were collected for the period 2013 through 2021 and are reported on a calendar year basis. Using production statistics from USDA:NASS as a denominator, the data supplied by participating companies represented approximately 82.1% of broiler chicken production in the U.S. in 2013, approximately 88.6% in 2017, and approximately 85.0% in 2021. The data that were submitted for 2021 are based on approximately 7,826,121,178 chickens slaughtered and 50,550,817,859 pounds liveweight produced. Granular flock-level treatment records were available for 75-90% of the birds represented in the 2018-2021 dataset. There was no use of antimicrobials in the hatchery for the years 2020 and 2021. Medically important in-feed antimicrobial use decreased substantially, with all in-feed tetracycline use being eliminated by 2020, and the use of virginiamycin being reduced by more than 97% since 2013. Medically important water-soluble antimicrobials are used for the treatment of disease in broiler production. Use decreased substantially for most water-soluble antimicrobials. The most important diseases necessitating treatment were necrotic enteritis and gangrenous dermatitis as well as E. coli-related disease. A focus on reducing the incidence of these diseases would reduce the need for antimicrobial therapy but will require an investment in research to find efficacious and cost-effective interventions for these diseases.
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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.
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When considering the development of antimicrobial resistance in food animals, comparing gross use estimates of different antimicrobials is of little value due to differences in potencies, duration of activity, relative effect on target and commensal bacteria, and mechanisms of resistance. However, it may be valuable to understand quantities of different antimicrobials used in different ages of swine and for what applications. Therefore, the objective of this project was to construct an estimate of antimicrobial use through the feed in swine production in the United States. Estimates were based on data from the National Animal Health Monitoring System (NAHMS) Swine 2006 Study and from a 2009 survey of swine-exclusive practitioners. Inputs consisted of number of pigs in a production phase, feed intake per day, dose of the antimicrobial in the feed, and duration of administration. Calculations were performed for a total of 102 combinations of antimicrobials (n=17), production phases (n=2), and reasons for use (n=3). Calculations were first conducted on farm-level data, and then extrapolated to the U.S. swine population. Among the nursery phase estimates, chlortetracycline had the largest estimate of use, followed by oxytetracycline and tilmicosin. In the grower/finisher phase, chlortetracycline also had the largest use estimate, followed by tylosin and oxytetracycline. As an annual industry estimate for all phases, chlortetracycline had the highest estimated use at 533,973 kg. The second and third highest estimates were tylosin and oxytetracycline with estimated annual uses of 165,803 kg and 154,956 kg, respectively. The estimates presented here were constructed to accurately reflect available data related to production practices, and to provide an example of a scientific approach to estimating use of compounds in production animals.
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Ração Animal/análise , Criação de Animais Domésticos/métodos , Anti-Infecciosos/administração & dosagem , Padrões de Prática Médica , Doenças dos Suínos/prevenção & controle , Médicos Veterinários , Drogas Veterinárias/administração & dosagem , Animais , Anti-Infecciosos/uso terapêutico , Clortetraciclina/administração & dosagem , Clortetraciclina/uso terapêutico , Doenças Transmitidas por Alimentos/prevenção & controle , Modelos Estatísticos , Oxitetraciclina/administração & dosagem , Oxitetraciclina/uso terapêutico , Inquéritos e Questionários , Sus scrofa , Suínos , Doenças dos Suínos/tratamento farmacológico , Fatores de Tempo , Tilosina/administração & dosagem , Tilosina/análogos & derivados , Tilosina/uso terapêutico , Estados Unidos , Drogas Veterinárias/uso terapêuticoRESUMO
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.
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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.
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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.
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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.
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Bovinos/sangue , Ensaio de Imunoadsorção Enzimática/veterinária , Interleucina-1beta/sangue , Fator de Necrose Tumoral alfa/sangue , Animais , Complexo Respiratório Bovino/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Interleucina-6/sangue , Estudos Longitudinais , Sensibilidade e EspecificidadeRESUMO
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.
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A prospective cohort observational study was conducted to investigate the seroprevalence of Anaplasma marginale in Iowa feedlots and its association with morbidity, mortality, and treatment costs. Blood samples were taken from 659 calves from 31 consigners at processing and classified as seropositive to A. marginale using a competitive enzyme-linked immunosorbent assay (cELISA) with a 30% cutoff. Health and production parameters were modeled by A. marginale serostatus with mixed model regression analysis. The apparent prevalence of seropositive cattle was 15.17% (100/659). When the cELISA positive cutoff was at 42% inhibition, the apparent prevalence was 5.00% (33/659). There was no significant association between A. marginale serostatus and production parameters; however, seropositive status had a weak positive association with undifferentiated fever (P = 0.17). Although prevalence of anaplasmosis in Iowa feedlots is higher than reported in Montana-sourced calves arriving in Canadian feedlots, this was not associated with increased production costs.
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Anaplasma marginale/imunologia , Anaplasmose/epidemiologia , Anticorpos Antibacterianos/sangue , Doenças dos Bovinos/epidemiologia , Anaplasmose/tratamento farmacológico , Anaplasmose/economia , Anaplasmose/prevenção & controle , Animais , Bovinos , Doenças dos Bovinos/tratamento farmacológico , Doenças dos Bovinos/economia , Doenças dos Bovinos/prevenção & controle , Estudos de Coortes , Custos e Análise de Custo , Iowa/epidemiologia , Estudos Prospectivos , Estudos SoroepidemiológicosRESUMO
Antimicrobial use is a key selective force behind the emergence of resistant bacteria. Therefore, optimizing strategies for more efficacious and targeted antimicrobial use is an essential component of efforts to combat antimicrobial resistance. To bolster stewardship programmes in animal agriculture, processes are needed for the systematic collection of on-farm antimicrobial use data. The objective of this study was to develop a system for collecting on-farm antimicrobial use data from the US broiler industry and to have it be representative of the largest commercial broiler producers in the United States that comprise the vast majority of national broiler production. Participation was voluntary. Data were collected for the period 2013 through 2017 and are reported on a calendar year basis. Using statistics from USDA:NASS as a denominator, the data supplied by participating companies accounted for approximately 81.7% of broiler production in the United States in 2013 and increased to approximately 87.2% in 2017. The data that were submitted for 2017 are based on approximately 7,897,339,357 chicks placed, 7,541,449,430 chickens slaughtered and 48,225,124,865 pounds liveweight produced. The use of antimicrobials in the hatchery decreased substantially between 2013 and 2017; the approximate percentage of broiler chicks placed that received hatchery antimicrobials decreased from 93% in 2013 to 17% in 2017. Medically important in-feed antimicrobial use decreased substantially. For example, in-feed tetracycline use decreased approximately 95% between 2013 and 2017. Medically important water-soluble antimicrobial use decreased substantially for most antimicrobials. Between 2013 and 2017, water-soluble penicillin use decreased approximately 21%, water-soluble tetracycline use decreased approximately 47%, and water-soluble lincomycin use decreased approximately 28%. While a reduction in antimicrobial amounts used may be an important indicator of improved stewardship, reducing the need for antimicrobials through improved disease prevention should be considered a more important objective and a better indicator of overall flock health and optimal antimicrobial use.