Departamento de Bacteriología y Virología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
Veterinario de Libre Ejercicio, Montevideo, Uruguay.
Appl Environ Microbiol. 2022 Jan 25;88(2):e0167521. doi: 10.1128/AEM.01675-21. Epub 2021 Nov 3.
Antimicrobial resistance is a critical issue that is no longer restricted to hospital settings but also represents a growing problem involving intensive animal production systems. In this study, we performed a microbiological and molecular investigation of priority pathogens carrying transferable resistance genes to critical antimicrobials in 1-day-old chickens imported from Brazil to Uruguay. Bacterial identification was performed by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry, and antibiotic susceptibility was determined by Sensititre. Antimicrobial resistance genes were sought by PCR, and clonality was assessed by pulsed-field gel electrophoresis (PFGE). Four multidrug-resistant (MDR) representative strains were sequenced by an Illumina and/or Oxford Nanopore Technologies device. Twenty-eight MDR isolates were identified as Escherichia coli ( 14), Enterobacter cloacae ( 11), or Klebsiella pneumoniae ( 3). While resistance to oxyiminocephalosporins was due to , , , , and , plasmid-mediated quinolone resistance was associated with the , , and genes. Finally, resistance to aminoglycosides and fosfomycin was due to the presence of 16S rRNA methyltransferase and -type genes, respectively. Short- and long-read genome sequencing of E. cloacae strain ODC_Eclo3 revealed the presence of IncQ/ (pUR-EC3.1; 7,400 bp), IncHI2A// (pUR-EC3.2, ST16 [pMLST; 408,436 bp), and IncN2/// (pUR-EC3.3) resistance plasmids. Strikingly, the gene was carried by a novel Tn-like composite transposon designated Tn. In summary, we report that imported 1-day-old chicks can act as Trojan horses for the hidden spread of WHO critical-priority MDR pathogens harboring , , and extended-spectrum β-lactamase genes in poultry farms, which is a critical issue from a One Health perspective. Antimicrobial resistance is considered a significant problem for global health, including within the concept of One Health; therefore, the food chain connects human health and animal health directly. In this work, we searched for microorganisms resistant to antibiotics considered critical for human health in intestinal microbiota of 1-day-old baby chicks imported to Uruguay from Brazil. We describe genes for resistance to antibiotics whose use the WHO has indicated to "watch" or "reserve" (AWaRe classification), such as and , which confer resistance to all the aminoglycosides and colistin, respectively, among other genes, and their presence in new mobile genetic elements that favor its dissemination. The sustained entry of these microorganisms evades the sanitary measures implemented by the countries and production establishments to reduce the selection of resistant microorganisms. These silently imported resistant microorganisms could explain a considerable part of the antimicrobial resistance problems found in the production stages of the system.
抗微生物药物耐药性不仅是医院环境中一个亟待解决的问题,也日益成为集约化动物生产系统中的一个严峻挑战。本研究对从巴西进口到乌拉圭的 1 日龄雏鸡中可转移的抗关键抗菌药物的耐药基因的优先病原体进行了微生物学和分子学调查。采用基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱法进行细菌鉴定,采用 Sensititre 进行抗生素药敏试验。通过 PCR 寻找抗微生物药物耐药基因,并通过脉冲场凝胶电泳(PFGE)评估克隆性。采用 Illumina 和/或牛津纳米孔技术设备对 4 株多药耐药(MDR)代表性菌株进行测序。鉴定出 28 株 MDR 分离株为大肠埃希菌(14 株)、阴沟肠杆菌(11 株)或肺炎克雷伯菌(3 株)。对氧肟头孢菌素的耐药性归因于 blaCTX-M-15、blaTEM-1B、blaDHA-1、blaSHV-12 和 qnrS1,而质粒介导的喹诺酮耐药性与 qnrS、qepA 和 aac(6')-Ib-cr 基因有关。最后,氨基糖苷类和磷霉素耐药性分别归因于 16S rRNA 甲基转移酶和 -型基因的存在。阴沟肠杆菌 ODC_Eclo3 菌株的短读长和长读长基因组测序揭示了 IncQ/(pUR-EC3.1;7400 bp)、IncHI2A//(pUR-EC3.2,ST16 [pMLST;408436 bp])和 IncN2///(pUR-EC3.3)耐药质粒的存在。值得注意的是, 基因位于一个新的命名为 Tn 的 Tn 样复合转座子上。综上所述,我们报告称,进口的 1 日龄雏鸡可作为隐藏传播世界卫生组织优先考虑的抗药性微生物(携带 和 以及扩展谱β-内酰胺酶基因)的特洛伊木马,这从一个大健康的角度来看是一个关键问题。抗微生物药物耐药性被认为是全球健康的一个重大问题,包括在大健康概念中;因此,食物链直接将人类健康和动物健康联系起来。在这项工作中,我们在从巴西进口到乌拉圭的 1 日龄小鸡的肠道微生物群中寻找被认为对人类健康至关重要的抗生素耐药微生物。我们描述了对 WHO 已指示“关注”或“保留”(AWaRe 分类)的抗生素的耐药基因,如 blaCTX-M-15、blaTEM-1B、blaDHA-1、blaSHV-12 和 qnrS1,这些基因分别赋予对所有氨基糖苷类和黏菌素的耐药性,以及其他基因,以及有利于其传播的新移动遗传元件。这些持续输入的耐药微生物逃避了各国和生产机构为减少耐药微生物选择而实施的卫生措施。这些被悄悄进口的耐药微生物可能可以解释在该系统的生产阶段发现的部分抗微生物药物耐药性问题。