Early childhood caries (ECC) is a severe manifestation of carious pathology with rapid and disruptive progression. The ECC microbiota includes a wide variety of bacterial species, among which is an anaerobic newly named species, Scardovia wiggsiae, a previously unidentified Bifidobacterium. Our aim was to provide the first ultrastructural characterization of S. wiggsiae and its biofilm by scanning electron microscopy (SEM) using a protocol that faithfully preserved the biofilm architecture and allowed an investigation at very high magnifications (order of nanometers) and with the appropriate resolution. To accomplish this task, we analyzed Streptococcus mutans’ biofilm by conventional SEM and VP-SEM protocols, in addition, we developed an original procedure, named OsO₄-RR-TA-IL, which avoids dehydration, drying and sputter coating. This innovative protocol allowed high-resolution and high-magnification imaging (from 10000× to 35000×) in high-vacuum and high-voltage conditions. After comparing three methods, we chose OsO₄-RR-TA-IL to investigate S. wiggsiae. It appeared as a fusiform elongated bacterium, without surface specialization, arranged in clusters and submerged in a rich biofilm matrix, which showed a well-developed micro-canalicular system. Our results provide the basis for the development of innovative strategies to quantify the effects of different treatments, in order to establish the best option to counteract ECC in pediatric patients.

中文翻译：

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通过原始扫描电子显微镜协议对金龟子生物膜进行表征。

幼儿龋齿（ECC）是龋齿病理的一种严重表现，具有快速而破坏性的进展。ECC微生物群包括各种各样的细菌，其中包括一个厌氧的新命名种，Scardovia wiggsiae ，一种以前未鉴定的双歧杆菌。我们的目的是通过扫描电子显微镜（SEM）使用忠实保留生物膜结构的协议，并以很高的放大倍数（纳米级）进行研究并提供适当分辨率，通过扫描电子显微镜（SEM）提供金黄色葡萄球菌及其生物膜的首个超微结构表征。。为了完成这个任务，我们分析小号treptococcus链球菌通过常规SEM和VP-SEM协议制备生物膜，此外，我们开发了一种原始程序，名为OsO ₄ -RR-TA-IL，可避免脱水，干燥和溅射涂层。这一创新的协议允许在高真空和高压条件下进行高分辨率和高倍率成像（从10000x到35000x）。比较三种方法之后，我们选择的OsO ₄ -RR-TA-IL调查S. wiggsiae 。它表现为梭形细长细菌，没有表面专门化，成簇排列并浸没在丰富的生物膜基质中，显示出发达的微细管系统。我们的结果为开发创新策略以量化不同治疗的效果提供了基础，从而建立了抵消儿科患者ECC的最佳选择。