Dataset Information

Effects of Ceftiofur and Chlortetracycline on the Resistomes of Feedlot Cattle.

ABSTRACT: Treatment of food-producing animals with antimicrobial drugs (AMD) is controversial because of concerns regarding promotion of antimicrobial resistance (AMR). To investigate this concern, resistance genes in metagenomic bovine fecal samples during a clinical trial were analyzed to assess the impacts of treatment on beef feedlot cattle resistomes. Four groups of cattle were exposed, using a 2-by-2 factorial design, to different regimens of antimicrobial treatment. Injections of ceftiofur crystalline-free acid (a third-generation cephalosporin) were used to treat all cattle in treatment pens or only a single animal, and either chlortetracycline was included in the feed of all cattle in a pen or the feed was untreated. On days 0 and 26, respectively, pre- and posttrial fecal samples were collected, and resistance genes were characterized using shotgun metagenomics. Treatment with ceftiofur was not associated with changes to ?-lactam resistance genes. However, cattle fed chlortetracycline had a significant increase in relative abundance of tetracycline resistance genes. There was also an increase of an AMR class not administered during the study, which is a possible indicator of coselection of resistance genes. Samples analyzed in this study had previously been evaluated by culture characterization (Escherichia coli and Salmonella) and quantitative PCR (qPCR) of metagenomic fecal DNA, which allowed comparison of results with this study. In the majority of samples, genes that were selectively enriched through culture and qPCR were not identified through shotgun metagenomic sequencing in this study, suggesting that changes previously documented did not reflect changes affecting the majority of bacterial genetic elements found in the predominant fecal resistome.IMPORTANCE Despite significant concerns about public health implications of AMR in relation to use of AMD in food animals, there are many unknowns about the long- and short-term impact of common uses of AMD for treatment, control, and prevention of disease. Additionally, questions commonly arise regarding how to best measure and quantify AMR genes in relation to public health risks and how to determine which genes are most important. These data provide an introductory view of the utility of using shotgun metagenomic sequencing data as an outcome for clinical trials evaluating the impact of using AMD in food animals.

SUBMITTER: Weinroth MD

PROVIDER: S-EPMC6007121 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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Effects of Ceftiofur and Chlortetracycline on the Resistomes of Feedlot Cattle.

Weinroth Margaret D MD Scott H Morgan HM Norby Bo B Loneragan Guy H GH Noyes Noelle R NR Rovira Pablo P Doster Enrique E Yang Xiang X Woerner Dale R DR Morley Paul S PS Belk Keith E KE

Applied and environmental microbiology 20180618 13

Treatment of food-producing animals with antimicrobial drugs (AMD) is controversial because of concerns regarding promotion of antimicrobial resistance (AMR). To investigate this concern, resistance genes in metagenomic bovine fecal samples during a clinical trial were analyzed to assess the impacts of treatment on beef feedlot cattle resistomes. Four groups of cattle were exposed, using a 2-by-2 factorial design, to different regimens of antimicrobial treatment. Injections of ceftiofur crystall ...[more]

PMID: 29728379

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Project description:Antibiotic use in beef cattle is a risk factor for the expansion of antimicrobial-resistant Salmonella populations. However, actual changes in the quantity of Salmonella in cattle feces following antibiotic use have not been investigated. Previously, we observed an overall reduction in Salmonella prevalence in cattle feces associated with both ceftiofur crystalline-free acid (CCFA) and chlortetracycline (CTC) use; however, during the same time frame the prevalence of multidrug-resistant Salmonella increased. The purpose of this analysis was to quantify the dynamics of Salmonella using colony counting (via a spiral-plating method) and hydrolysis probe-based qPCR (TaqMan® qPCR). Additionally, we quantified antibiotic-resistant Salmonella by plating to agar containing antibiotics at Clinical & Laboratory Standards Institute breakpoint concentrations. Cattle were randomly assigned to 4 treatment groups across 16 pens in 2 replicates consisting of 88 cattle each. Fecal samples from Days 0, 4, 8, 14, 20, and 26 were subjected to quantification assays. Duplicate qPCR assays targeting the Salmonella invA gene were performed on total community DNA for 1,040 samples. Diluted fecal samples were spiral plated on plain Brilliant Green Agar (BGA) and BGA with ceftriaxone (4 ?g/ml) or tetracycline (16 ?g/ml). For comparison purposes, indicator non-type-specific (NTS) E. coli were also quantified by direct spiral plating. Quantity of NTS E. coli and Salmonella significantly decreased immediately following CCFA treatment. CTC treatment further decreased the quantity of Salmonella but not NTS E. coli. Effects of antibiotics on the imputed log10 quantity of Salmonella were analyzed via a multi-level mixed linear regression model. The invA gene copies decreased with CCFA treatment by approximately 2 log10 gene copies/g feces and remained low following additional CTC treatment. The quantities of tetracycline or ceftriaxone-resistant Salmonella were approximately 4 log10 CFU/g feces; however, most of the samples were under the quantification limit. The results of this study demonstrate that antibiotic use decreases the overall quantity of Salmonella in cattle feces in the short term; however, the overall quantities of antimicrobial-resistant NTS E. coli and Salmonella tend to remain at a constant level throughout.

Project description:A randomized controlled field trial was conducted to evaluate the effects of two sets of treatment strategies on ceftiofur and tetracycline resistance in feedlot cattle. The strategies consisted of ceftiofur crystalline-free acid (CCFA) administered to either one or all of the steers within a pen, followed by feeding or not feeding a therapeutic dose of chlortetracycline (CTC). Eighty-eight steers were randomly allocated to eight pens of 11 steers each. Both treatment regimens were randomly assigned to the pens in a two-way full factorial design. Non-type-specific (NTS) E. coli (n = 1,050) were isolated from fecal samples gathered on Days 0, 4, 12, and 26. Antimicrobial susceptibility profiles were determined using a microbroth dilution technique. PCR was used to detect tet(A), tet(B), and bla CMY-2 genes within each isolate. Chlortetracycline administration greatly exacerbated the already increased levels of both phenotypic and genotypic ceftiofur resistance conferred by prior CCFA treatment (P<0.05). The four treatment regimens also influenced the phenotypic multidrug resistance count of NTS E. coli populations. Chlortetracycline treatment alone was associated with an increased probability of selecting isolates that harbored tet(B) versus tet(A) (P<0.05); meanwhile, there was an inverse association between finding tet(A) versus tet(B) genes for any given regimen (P<0.05). The presence of a tet(A) gene was associated with an isolate exhibiting reduced phenotypic susceptibility to a higher median number of antimicrobials (n = 289, median = 6; 95% CI = 4-8) compared with the tet(B) gene (n = 208, median = 3; 95% CI = 3-4). Results indicate that CTC can exacerbate ceftiofur resistance following CCFA therapy and therefore should be avoided, especially when considering their use in sequence. Further studies are required to establish the animal-level effects of co-housing antimicrobial-treated and non-treated animals together.

Project description:Concerns have been raised that in-feed chlortetracycline (CTC) may increase antimicrobial resistance (AMR), specifically tetracycline-resistant (TETr) Escherichia coli and third-generation cephalosporin-resistant (3GCr) E. coli We evaluated the impact of a 5-day in-feed CTC prophylaxis on animal health, TETr E. coli, and 3GCr E. coli A control group of cattle (n = 150) received no CTC, while a CTC group (n = 150) received in-feed CTC (10 mg/lb of body weight/day) from the 5th to the 9th day after feedlot arrival. Over 25% (38/150) of the animals in the control group developed illnesses requiring therapeutic treatment with antimicrobials critically important to human medicine. Only two animals (1.3%) in the CTC group required such treatments. Fecal swab and pen surface occurrences of generic E. coli (isolated on media that did not contain antimicrobials of interest and were not isolated based on any specific resistance), TETr E. coli, and 3GCr E. coli were determined on five sampling occasions: arrival at the feedlot, 5 days posttreatment (5 dpt), 27 dpt, 75 dpt, and 117 dpt. On 5 dpt, TETr E. coli concentrations were higher for the CTC group than the control group (P < 0.01). On 27 dpt, 75 dpt, and 117 dpt, TETr E. coli concentrations did not differ between groups. 3GCr E. coli occurrences did not differ between control and CTC groups on any sampling occasion. For both groups, generic, TETr, and 3GCr E. coli occurrences were highest on 75 dpt and 117 dpt, suggesting that factors other than in-feed CTC contributed more significantly to antimicrobial-resistant E. coli occurrence.ImportanceThe occurrence of human bacterial infections resistant to antimicrobial therapy has been increasing. It has been postulated that antimicrobial resistance was inevitable, but the life span of the antimicrobial era has been prematurely compromised due to the misuse of antimicrobials in clinical and agricultural practices. Direct evidence relating the use of antimicrobials in livestock production to diminished human health outcomes due to antimicrobial resistance is lacking, and the U.S. Food and Drug Administration has taken an approach to maximize therapeutic efficacy and minimize the selection of resistant microorganisms through judicious use of antimicrobials. This study demonstrated that prophylactic in-feed treatment of chlortetracycline administered for 5 days to calves entering feedlots is judicious, as this therapy reduced animal morbidity, reduced the use of antimicrobials more critical to human health, and had no long-term impact on the occurrence of antimicrobial-resistant E. coli.

Project description:The objective was to determine effects of old and new rubber matting in a slatted, indoor cattle feeding facility on cattle growth performance, locomotion, and carcass characteristics. In experiment 1, fall-born Angus × Simmental steers (N = 207; body weight = 222 ± 38 kg) were blocked by weight and assigned to 32 pens. Pens were randomly assigned to 1 of 4 treatments: no matting/concrete (CONC1), 12-yr-old Animat Pebble matting (OLD1), new Animat Maxgrip matting (MG), and new Animat Pebble matting (PEB1). Steers were fed a common diet for 209 d with a minimum stocking density of 3.40 m2 per animal. Final body weight (BW) and average daily gain (ADG) were affected (P = 0.02 and P < 0.01, respectively) by treatment with steers on PEB1 finishing heaviest with the greatest growth, MG and CONC1 intermediate, and OLD1 finishing at the lightest final BW with the least growth. Flooring treatment did not affect overall dry matter intake (DMI; P = 0.16) or gain to feed ratio (G:F; P = 0.94). Flooring treatment did not affect (P ≥ 0.19) any carcass traits. Locomotion scores (LS) were affected (P < 0.01) by flooring treatment with CONC1 having the worst mobility while OLD1, MG, and PEB1 were similar (P ≥ 0.24). Locomotion score had a day effect (P < 0.01) where cattle gait and mobility worsened as days on feed increased. In experiment 2, fall-born Angus × Simmental steers (N = 189; BW = 352 ± 43 kg) were blocked by weight and assigned to 21 pens. Pens were randomly assigned to 1 of 3 treatments: no matting/concrete (CONC2), 15-yr-old Animat Pebble matting (OLD2), and new Animat Pebble matting (PEB2). Steers were fed a common diet for 152 d with a stocking density of 2.65 m2 per steer. After 152 d on feed, flooring treatment did not affect (P ≥ 0.30) BW, ADG, or DMI nor did treatment affect (P ≥ 0.17) carcass traits. However, steers housed on OLD2 or PEB2 had improved locomotion scores (P = 0.02) compared with steers housed on CONC2. Locomotion score had a day effect (P < 0.01) as cattle gait and mobility worsened with greater number of days on feed, regardless of treatment. Overall, results suggest that new rubber matting increased ADG and HCW during a 209-d trial when cattle were stocked at 3.4 m2 in small pens and that rubber matting regardless of age improved cattle locomotion scores in slatted indoor feeding facilities.

Dataset Information

Effects of Ceftiofur and Chlortetracycline on the Resistomes of Feedlot Cattle.

Publications

Effects of Ceftiofur and Chlortetracycline on the Resistomes of Feedlot Cattle.

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