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短篇通讯:不同临床型乳房炎严重度评分奶牛源致病性大肠埃希氏菌的肠外致病性毒力基因、细菌运动性和多药耐药模式的调查。

Short communication: Investigation of extra-intestinal pathogenic Escherichia coli virulence genes, bacterial motility, and multidrug resistance pattern of strains isolated from dairy cows with different severity scores of clinical mastitis.

机构信息

Department of Veterinary Hygiene and Public Health, School of Veterinary Medicine and Animal Sciences, Sao Paulo State University, Botucatu, SP 18618 681, Brazil.

Department of Microbiology and Immunology, Sao Paulo State University, Botucatu, SP 18618 689, Brazil.

出版信息

J Dairy Sci. 2020 Apr;103(4):3606-3614. doi: 10.3168/jds.2019-17477. Epub 2020 Feb 7.

Abstract

Escherichia coli is a major pathogen involved in the etiology of environmentally derived bovine mastitis and is characterized by a variety of virulence factors (VF). Mammary infections with E. coli have shown a wide range of clinical signs, causing changes in milk (score 1, or mild), abnormal appearance of milk and udder inflammation (score 2, or moderate), and abnormalities in milk, udder inflammation, and systemic signs of illness (score 3, or severe). Nevertheless, to date, the profile of the genes related to the virulence of the pathogen in mammary infections and the severity scores of cases have not been thoroughly elucidated. Therefore, a panel of 18 virulence-encoding genes associated with extra-enteric pathogenicity of E. coli (ExPEC) were investigated in addition to in vitro swimming and swarming motility profiles and antimicrobial susceptibility/resistance patterns among 114 E. coli strains isolated from cows with clinical mastitis and different severity scores. Of 114 clinical cases, 39.5, 54.4, and 6.1% were mild, moderate, and severe, respectively. The main genes related to VF harbored by isolates were adhesins (fimH 100%; ecpA 64.0%, fimA 31.6%), serum resistance (traT 81.6%; ompT 35.1%), siderophores (irp2 9.6%), and hemolysin (hlyA 7%). Among the isolates studied, 99.1% showed in vitro resistance to bacitracin and cloxacillin, and 98.2% to lincosamin. Of the total isolates, 98.2% were considered multidrug resistant based on the multiple antimicrobial resistance index. No significant difference was observed between mean swimming (13.8 mm) and swarming (13.5 mm) motility, as well as severity scores of clinical mastitis and the ExPEC genes studied. The isolation of strains resistant to various antimicrobials, even though tested only in vitro, highlights the importance of rational use of antimicrobials for mastitis treatment. The high prevalence of the genes related to serum resistance (traT and ompT) and adhesion (ecpA) of the pathogen, in addition to main associations between the genes fimH, ecpA, and traT among cows with severity scores of 1 (15%) and 2 (22.6%), indicates that the genes traT, ecpA, and ompT could be further studied as biomarkers of ExPEC for clinical intramammary infections. In addition, the ExPEC genes ompT (protectin), ibe10 (invasin), and ecpA (adhesin) were investigated for the first time among cows with mastitis, where scores of clinical severity were assessed. Results of this study contribute to the characterization of virulence mechanisms and antimicrobial resistance profile of ExPEC variants that affect dairy cows with different scores of clinical mastitis.

摘要

大肠杆菌是一种主要的病原体,参与了环境源性牛乳腺炎的病因学,其特征是多种毒力因子(VF)。大肠杆菌引起的乳房感染表现出广泛的临床症状,导致乳汁变化(评分 1,或轻度)、乳汁外观异常和乳房炎症(评分 2,或中度)以及乳汁、乳房炎症和全身疾病体征异常(评分 3,或重度)。然而,迄今为止,与乳腺感染中病原体毒力相关的基因谱以及病例严重程度评分尚未得到彻底阐明。因此,除了体外游泳和群集运动特征外,还研究了与大肠杆菌(ExPEC)肠外致病性相关的 18 个毒力编码基因,以及 114 株从患有不同严重程度临床乳腺炎的奶牛中分离的大肠杆菌菌株的抗菌药物敏感性/耐药模式。在 114 例临床病例中,轻度、中度和重度分别占 39.5%、54.4%和 6.1%。分离株携带的主要与 VF 相关的基因是黏附素( fimH 100%; ecpA 64.0%, fimA 31.6%)、血清抗性( traT 81.6%; ompT 35.1%)、铁载体( irp2 9.6%)和溶血性( hlyA 7%)。在所研究的分离株中,99.1%对杆菌肽和氯唑西林具有体外耐药性,98.2%对林可酰胺耐药。基于多重抗菌药物耐药指数,总分离株中有 98.2%被认为是多药耐药。大肠杆菌的平均游泳(13.8 毫米)和群集运动(13.5 毫米)以及临床乳腺炎严重程度评分与 ExPEC 基因之间没有观察到显著差异。尽管仅在体外进行了测试,但对各种抗菌药物的耐药菌株的分离突出了合理使用抗菌药物治疗乳腺炎的重要性。血清抗性( traT 和 ompT)和黏附( ecpA)相关基因以及 fimH、 ecpA 和 traT 基因之间的主要关联在严重程度评分分别为 1(15%)和 2(22.6%)的奶牛中高度流行,表明基因 traT、 ecpA 和 ompT 可以进一步作为临床乳腺内感染的 ExPEC 生物标志物进行研究。此外,首次在患有乳腺炎且评估了临床严重程度评分的奶牛中研究了 ExPEC 基因 ompT(保护素)、 ibe10(入侵素)和 ecpA(黏附素)。本研究的结果有助于表征不同严重程度临床乳腺炎奶牛中影响 ExPEC 变体的毒力机制和抗菌药物耐药谱。

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