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猪分离株的表型和基因型特征

The Pheno- and Genotypic Characterization of Porcine Isolates.

作者信息

Bernreiter-Hofer Tanja, Schwarz Lukas, Müller Elke, Cabal-Rosel Adriana, Korus Maciej, Misic Dusan, Frankenfeld Katrin, Abraham Kerstin, Grünzweil Olivia, Weiss Astrid, Feßler Andrea T, Allerberger Franz, Schwarz Stefan, Szostak Michael P, Ruppitsch Werner, Ladinig Andrea, Spergser Joachim, Braun Sascha D, Monecke Stefan, Ehricht Ralf, Loncaric Igor

机构信息

Institute of Microbiology, University of Veterinary Medicine, 1210 Vienna, Austria.

Department for Farm Animals and Veterinary Public Health, University Clinic for Swine, University of Veterinary Medicine, 1210 Vienna, Austria.

出版信息

Microorganisms. 2021 Aug 6;9(8):1676. doi: 10.3390/microorganisms9081676.

Abstract

() is the main causative pathogen of neonatal and post-weaning diarrhea and edema disease in swine production. There is a significant health concern due to an increasing number of human infections associated with food and/or environmental-borne pathogenic and multidrug-resistant worldwide. Monitoring the presence of pathogenic and antimicrobial-resistant isolates is essential for sustainable disease management in livestock and human medicine. A total of 102 isolates of diseased pigs were characterized by antimicrobial and biocide susceptibility testing. Antimicrobial resistance genes, including mobile colistin resistance genes, were analyzed by PCR and DNA sequencing. The quinolone resistance-determining regions of and in ciprofloxacin-resistant isolates were analyzed. Clonal relatedness was investigated by two-locus sequence typing (CH clonotyping). Phylotyping was performed by the Clermont multiplex PCR method. Virulence determinants were analyzed by customized DNA-based microarray technology developed in this study for fast and economic molecular multiplex typing. Thirty-five isolates were selected for whole-genome sequence-based analysis. Most isolates were resistant to ampicillin and tetracycline. Twenty-one isolates displayed an ESBL phenotype and one isolate an AmpC β-lactamase-producing phenotype. Three isolates had elevated colistin minimal inhibitory concentrations and carried the gene. Thirty-seven isolates displayed a multi-drug resistance phenotype. The most predominant β-lactamase gene classes were (56%) and (13.71%). Mutations in QRDR were observed in 14 ciprofloxacin-resistant isolates. CH clonotyping divided all isolates into 51 CH clonotypes. The majority of isolates belonged to phylogroup A. Sixty-four isolates could be assigned to defined pathotypes wherefrom UPEC was predominant. WGS revealed that the most predominant sequence type was ST100, followed by ST10. ST131 was detected twice in our analysis. This study highlights the importance of monitoring antimicrobial resistance and virulence properties of porcine isolates. This can be achieved by applying reliable, fast, economic and easy to perform technologies such as DNA-based microarray typing. The presence of high-risk pathogenic multi-drug resistant zoonotic clones, as well as those that are resistant to critically important antibiotics for humans, can pose a risk to public health. Improved protocols may be developed in swine farms for preventing infections, as well as the maintenance and distribution of the causative isolates.

摘要

()是养猪生产中新生仔猪和断奶后腹泻及水肿病的主要致病病原体。由于全球范围内与食物和/或环境传播的致病性及多重耐药性相关的人类感染病例不断增加,这引发了重大的健康担忧。监测致病性和抗菌药物耐药性分离株的存在对于家畜和人类医学的可持续疾病管理至关重要。通过抗菌药物和杀生物剂敏感性测试对102株患病猪的分离株进行了特征分析。通过聚合酶链反应(PCR)和DNA测序分析了包括移动性黏菌素耐药基因在内的抗菌药物耐药基因。对环丙沙星耐药分离株中的和的喹诺酮耐药决定区进行了分析。通过双位点序列分型(CH克隆分型)研究了克隆相关性。通过克莱蒙特多重聚合酶链反应方法进行系统发育分型。通过本研究开发的定制基于DNA的微阵列技术分析毒力决定因素,以实现快速且经济的分子多重分型。选择了35株分离株进行基于全基因组序列的分析。大多数分离株对氨苄青霉素和四环素耐药。21株分离株表现出超广谱β-内酰胺酶(ESBL)表型,1株分离株表现出产AmpCβ-内酰胺酶表型。3株分离株的黏菌素最低抑菌浓度升高并携带基因。37株分离株表现出多重耐药表型。最主要的β-内酰胺酶基因类别是(56%)和(13.71%)。在14株环丙沙星耐药分离株中观察到喹诺酮耐药决定区(QRDR)的突变。CH克隆分型将所有分离株分为51个CH克隆型。大多数分离株属于A系统发育群。64株分离株可归为确定的致病型,其中以尿路致病性大肠埃希菌(UPEC)为主。全基因组测序显示最主要的序列型是ST100,其次是ST10。在我们的分析中检测到ST131两次。本研究强调了监测猪分离株的抗菌药物耐药性和毒力特性的重要性。这可以通过应用可靠、快速、经济且易于操作的技术如基于DNA的微阵列分型来实现。高风险致病性多重耐药人畜共患病克隆以及对人类至关重要的抗生素耐药的克隆的存在可能对公共卫生构成风险。养猪场可能需要制定改进方案以预防感染以及致病分离株的保存和传播。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa61/8400056/3c8f0d1d60fd/microorganisms-09-01676-g001.jpg

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