Silvestre Inês, Borrego Maria José, Jordão Luísa
Department of Life Sciences, UCIBIO, Nova School of Science and Technology, 2829-516 Caparica, Portugal; Department of Infectious Diseases, National Institute of Health Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016 Lisbon, Portugal.
Department of Infectious Diseases, National Institute of Health Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016 Lisbon, Portugal.
Res Microbiol. 2020 Dec;171(8):311-318. doi: 10.1016/j.resmic.2020.08.001. Epub 2020 Sep 5.
Bacterial biofilms are an important virulence factor with a vital role in evasion from the host immune system, colonization and infection. The aim of the present study was to evaluate in vitro the effects of three environmental factors (H, glucose and human plasma) in biofilm formation, by carrier and invasive Streptococcus agalactiae strains of ST17 and ST19 sequence types, including DNase producers and non-producers. Bacteria ability to assemble biofilms was classified based on crystal violet assay. Biofilm formation was also monitored by scanning electron microscopy. Depending on the growth medium used, each bacterial isolate could fit in different biofilm production categories. Our data showed that optimal conditions for S. agalactiae biofilm assembly were reached after 48 h incubation at pH 7.6 in the presence of glucose and inactivated human plasma. In the presence of inactivated human plasma, the biofilm biomass of ST19 strains experienced a higher increase than ST17 strains. The composition of the extracellular polymeric matrix of the three strongest biofilm producers (all from ST17) was accessed by enzymatic digestion of mature biofilms and proteins were shown to be the predominant component. The detailed identification of the extracellular protein components should contribute to the development of new therapeutic strategies to fight S. agalactiae infections.
细菌生物膜是一种重要的毒力因子,在逃避宿主免疫系统、定植和感染过程中起着至关重要的作用。本研究旨在体外评估三种环境因素(氢离子浓度、葡萄糖和人血浆)对ST17和ST19序列类型的携带型和侵袭型无乳链球菌菌株生物膜形成的影响,这些菌株包括产DNA酶和不产DNA酶的菌株。基于结晶紫测定法对细菌形成生物膜的能力进行分类。还通过扫描电子显微镜监测生物膜的形成。根据所使用的生长培养基,每种细菌分离株可归为不同的生物膜产生类别。我们的数据表明,在pH 7.6、存在葡萄糖和灭活的人血浆的条件下孵育48小时后,可达到无乳链球菌生物膜组装的最佳条件。在存在灭活的人血浆的情况下,ST19菌株的生物膜生物量比ST17菌株有更高的增加。通过对成熟生物膜进行酶消化来研究三种最强生物膜产生菌(均来自ST17)的细胞外聚合物基质的组成,结果表明蛋白质是主要成分。对细胞外蛋白质成分的详细鉴定应有助于开发对抗无乳链球菌感染的新治疗策略。