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巴西奶牛乳房炎金黄色葡萄球菌分离株的生物膜形成及对β-内酰胺类抗生素的耐药性

Biofilm production and beta-lactamic resistance in Brazilian Staphylococcus aureus isolates from bovine mastitis.

作者信息

Marques Viviane Figueira, Motta Cássia Couto da, Soares Bianca da Silva, Melo Dayanne Araújo de, Coelho Shana de Mattos de Oliveira, Coelho Irene da Silva, Barbosa Helene Santos, Souza Miliane Moreira Soares de

机构信息

Universidade Federal Rural do Rio de Janeiro, Microbiologia e Imunologia Veterinária, Seropédica, Rio de Janeiro, RJ, Brazil.

Universidade Federal Rural do Rio de Janeiro, Microbiologia e Imunologia Veterinária, Instituto de Veterinária (IV), Seropédica, Rio de Janeiro, RJ, Brazil.

出版信息

Braz J Microbiol. 2017 Jan-Mar;48(1):118-124. doi: 10.1016/j.bjm.2016.10.001. Epub 2016 Oct 18.

Abstract

Staphylococcus spp. play an important role in the etiology of bovine mastitis. Staphylococcus aureus is considered the most relevant species due to the production of virulence factors such as slime, which is required for biofilm formation. This study aimed to evaluate biofilm production and its possible relation to beta-lactamic resistance in 20 S. aureus isolates from bovine mastitic milk. The isolates were characterized by pheno-genotypic and MALDI TOF-MS assays and tested for genes such as icaA, icaD, bap, agr RNAIII, agr I, agr II, agr III, and agr IV, which are related to slime production and its regulation. Biofilm production in microplates was evaluated considering the intervals determined along the bacterial growth curve. In addition, to determine the most suitable time interval for biofilm analysis, scanning electron microscopy was performed. Furthermore, genes such as mecA and blaZ that are related to beta-lactamic resistance and oxacillin susceptibility were tested. All the studied isolates were biofilm producers and mostly presented icaA and icaD. The Agr type II genes were significantly prevalent. According to the SEM, gradual changes in the bacterial arrangement were observed during biofilm formation along the growth curve phases, and the peak was reached at the stationary phase. In this study, the penicillin resistance was related to the production of beta-lactamase, and the high minimal bactericidal concentration for cefoxitin was possibly associated with biofilm protection. Therefore, further studies are warranted to better understand biofilm formation, possibly contributing to our knowledge about bacterial resistance in vivo.

摘要

葡萄球菌属在牛乳腺炎的病因学中起重要作用。金黄色葡萄球菌被认为是最相关的菌种,因为它能产生如黏液等毒力因子,而黏液是生物膜形成所必需的。本研究旨在评估从患乳腺炎的牛乳中分离出的20株金黄色葡萄球菌的生物膜形成情况及其与β-内酰胺耐药性的可能关系。通过表型-基因型和基质辅助激光解吸电离飞行时间质谱(MALDI TOF-MS)分析对分离株进行鉴定,并检测与黏液产生及其调控相关的基因,如icaA、icaD、bap、agr RNAIII、agr I、agr II、agr III和agr IV。根据细菌生长曲线确定的时间间隔,评估微孔板中的生物膜形成情况。此外,为确定生物膜分析的最合适时间间隔,进行了扫描电子显微镜检查。此外,还检测了与β-内酰胺耐药性和苯唑西林敏感性相关的基因,如mecA和blaZ。所有研究的分离株均为生物膜产生菌,且大多呈现icaA和icaD。Agr II型基因显著流行。根据扫描电子显微镜观察,在生物膜形成过程中,沿生长曲线各阶段细菌排列逐渐变化,在稳定期达到峰值。在本研究中,青霉素耐药性与β-内酰胺酶的产生有关,头孢西丁的高最小杀菌浓度可能与生物膜保护有关。因此,有必要进一步研究以更好地理解生物膜形成,这可能有助于我们了解体内细菌耐药性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd83/5221365/2f21d65e5cdd/gr1.jpg

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