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Global responses to oxytetracycline treatment in tetracycline-resistant Escherichia coli.

ABSTRACT: We characterized the global transcriptome of Escherichia coli MG1655:: tetA grown in the presence of ½ MIC (14?mg/L) of OTC, and for comparison WT MG1655 strain grown with 1//2 MIC of OTC (0.25?mg/L OTC). 1646 genes changed expression significantly (FDR?>?0.05) in the resistant strain, the majority of which (1246) were also regulated in WT strain. Genes involved in purine synthesis and ribosome structure and function were top-enriched among up-regulated genes, and anaerobic respiration, nitrate metabolism and aromatic amino acid biosynthesis genes among down-regulated genes. Blocking of the purine-synthesis- did not affect resistance phenotypes (MIC and growth rate with OTC), while blocking of protein synthesis using low concentrations of chloramphenicol or gentamicin, lowered MIC towards OTC. Metabolic-modeling, using a novel model for MG1655 and continuous weighing factor that reflected the degree of up or down regulation of genes encoding a reaction, identified 102 metabolic reactions with significant change in flux in MG1655:: tetA when grown in the presence of OTC compared to growth without OTC. These pathways could not have been predicted by simply analyzing functions of the up and down regulated genes, and thus this work has provided a novel method for identification of reactions which are essential in the adaptation to growth in the presence of antimicrobials.

SUBMITTER: Moller TSB

PROVIDER: S-EPMC7242477 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Global responses to oxytetracycline treatment in tetracycline-resistant Escherichia coli.

Møller Thea S B TSB Liu Gang G Hartman Hassan B HB Rau Martin H MH Mortensen Sisse S Thamsborg Kristian K Johansen Andreas E AE Sommer Morten O A MOA Guardabassi Luca L Poolman Mark G MG Olsen John E JE

Scientific reports 20200521 1

We characterized the global transcriptome of Escherichia coli MG1655:: tetA grown in the presence of ½ MIC (14 mg/L) of OTC, and for comparison WT MG1655 strain grown with 1//2 MIC of OTC (0.25 mg/L OTC). 1646 genes changed expression significantly (FDR > 0.05) in the resistant strain, the majority of which (1246) were also regulated in WT strain. Genes involved in purine synthesis and ribosome structure and function were top-enriched among up-regulated genes, and anaerobic respiration, nitrate ...[more]

PMID: 32439837

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Project description:BackgroundAntimicrobial helper-compounds may reverse antimicrobial resistance. Sertraline, a antidepressant drug, has been suggested as a tetracycline helper-compound. Tetracycline is the preferred antimicrobial for treatment of enteric diseases in pigs. This study is the first to evaluate the potency of sertraline as a tetracycline adjuvant in pigs.MethodsForty-eight nursery pigs were divided into four treatment groups: Tetracycline, sertraline, tetracycline/sertraline or un-medicated control. Fecal and ileal samples were obtained before treatment, 48?h and nine days after five days of treatment, respectively. Colony forming units (CFU) of tetracycline resistant coliforms in each sample (ileal or fecal) and CFU of an orally inoculated tetracycline-resistant strain of Escherichia coli were determined at each sampling point. The microbiome of fecal and ileal and samples was analyzed by sequencing of the 16S V3-V4 region.ResultsThe results did not provide evidence that sertraline in combination with tetracycline has any impact on tetracycline resistant bacteria in either fecal or ileum samples, while in the tetracycline treated group of pigs, an increase in the prevalence of a tetracycline resistant indicator strain of Escherichia coli shortly after ended five-day treatment was observed. The ileal samples obtained shortly after ended treatment showed treatment-associated changes in the composition of the microbiota in the groups of pigs treated with tetracycline (+/-) sertraline. While tetracycline treatment increased the abundance in the reads of E. coli, sertraline/tetracycline treatment led to increased abundances of Streptococcus spp. and decreased abundances of Lactobacillus spp. However, all observed differences (on CFU counts and microbiota composition) between groups shortly after treatment had diminished in less than two weeks after last treatment day.ConclusionsSertraline (+/-) tetracycline treatment did not reduce the long-term level of tetracycline-resistant bacteria in the feces or small intestine contents of piglets compared to the un-medicated control group of pigs. The result of this study reflects the importance of in vivo studies for confirmation of the antimicrobial helper-compound potential of an in vitro active compound.

Project description:BACKGROUND: Feedlot cattle in North America are routinely fed subtherapeutic levels of antimicrobials to prevent disease and improve the efficiency of growth. This practice has been shown to promote antimicrobial resistance (AMR) in subpopulations of intestinal microflora including Escherichia coli. To date, studies of AMR in feedlot production settings have rarely employed selective isolation, therefore yielding too few AMR isolates to enable characterization of the emergence and nature of AMR in E. coli as an indicator bacterium. E. coli isolates (n = 531) were recovered from 140 cattle that were housed (10 animals/pen) in 14 pens and received no dietary antimicrobials (control--5 pens, CON), or were intermittently administered subtherapeutic levels of chlortetracycline (5 pens-T), chlortetracycline + sulfamethazine (4 pens-TS), or virginiamycin (5 pens-V) for two separate periods over a 9-month feeding period. Phenotype and genotype of the isolates were determined by susceptibility testing and pulsed field gel electrophoresis and distribution of characterized isolates among housed cattle reported. It was hypothesized that the feeding of subtherapeutic antibiotics would increase the isolation of distinct genotypes of AMR E. coli from cattle. RESULTS: Overall, patterns of antimicrobial resistance expressed by E. coli isolates did not change among diet groups (CON vs. antibiotic treatments), however; isolates obtained on selective plates (i.e., M(A),M(T)), exhibited multi-resistance to sulfamethoxazole and chloramphenicol more frequently when obtained from TS-fed steers than from other treatments. Antibiograms and PFGE patterns suggested that AMR E. coli were readily transferred among steers within pens. Most M(T) isolates possessed the tet(B) efflux gene (58.2, 53.5, 40.8, and 50.6% of isolates from CON, T, TS, and V steers, respectively) whereas among the M(A) (ampicillin-resistant) isolates, the tem1-like determinant was predominant (occurring in 50, 66.7, 80.3, and 100% of isolates from CON, T, TS, and V steers, respectively). CONCLUSIONS: Factors other than, or in addition to subtherapeutic administration of antibiotics influence the establishment and transmission of AMR E. coli among feedlot cattle.

Project description:Antimicrobial resistance (AMR) is an emerging global concern, with the widespread use of antimicrobials in One Health contributing significantly to this phenomenon. Among various antimicrobials, tetracyclines are extensively used in the beef cattle industry, potentially contributing to the development of resistance in bacterial populations. This meta-analysis aimed to examine the association between tetracycline use in beef cattle and the development of tetracycline resistance in Escherichia coli isolates. A comprehensive search was conducted using multiple databases to gather relevant observational studies evaluating tetracycline use and tetracycline resistance in Escherichia coli isolates from beef cattle. The rate of tetracycline resistance from each study served as the effect measure and was pooled using a random-effects model, considering possible disparities among studies. The meta-analysis of 14 prospective longitudinal studies resulted in a 0.31 prevalence of tetracycline resistance in Escherichia coli in non-intervention (no exposure), contrasting numerically elevated resistance rates in the intervention (exposed) groups of 0.53 and 0.39 in those receiving tetracyclines via feed or systemically, respectively. Despite the observed numerical differences, no statistically significant differences existed between intervention and non-intervention groups, challenging the conventional belief that antimicrobial use in livestock inherently leads to increased AMR. The findings of this study underscore the need for additional research to fully understand the complex relationship between antimicrobial use and AMR development. A considerable degree of heterogeneity across studies, potentially driven by variations in study design and diverse presentation of results, indicates the intricate and complex nature of AMR development. Further research with standardized methodologies might help elucidate the relationship between tetracycline use and resistance in Escherichia coli isolated from beef cattle.

Dataset Information

Global responses to oxytetracycline treatment in tetracycline-resistant Escherichia coli.

Publications

Global responses to oxytetracycline treatment in tetracycline-resistant Escherichia coli.

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