Lactobacilli, the most common group of bacteria found in a healthy vaginal microbiota, have been demonstrated to act as a defence against colonisation and overgrowth of vaginal pathogens. These groups of bacteria have sparked interests in incorporating them as probiotics aimed at re-establishing balance within the urogenital ecosystem. In this study, the safety characteristics of Limosilactobacillus reuteri 29B (L29B) strain were evaluated through whole genome sequencing (WGS) and animal study. Cell culture assay and 16S rDNA analysis were done to evaluate the ability of the strain to colonise and adhere to the mouse vaginal tract, and RAST analysis was performed to screen for potential genes associated with probiotic trait. The histological study on the mice organs and blood analysis of the mice showed there was no incidence of inflammation. We also found no evidence of bacterial translocation. The cell culture assay on HeLa cells showed 85% of adhesion, and there was a significant reduction of Candida strain viability in displacement assay. As for the 16S rDNA analysis, there was a significant amount of L29B colonisation of the vaginal microflora. Taken together, the intravaginal administration of L29B significantly reduced the number Enterobacteriaceae and Staphylococcaceae that were present in mouse vaginal tract. It also improved and promoted a balanced vaginal microflora environment without causing any harm or irritation to mice. Limosilactobacillus 29B (L29B) is safe to be administered intravaginally.
Candida glabrata is an emerging human fungal pathogen that has efficacious nutrient sensing and responsiveness ability. It can be seen through its ability to thrive in diverse range of nutrient limited-human anatomical sites. Therefore, nutrient sensing particularly glucose sensing is thought to be crucial in contributing to the development and fitness of the pathogen. This study aimed to elucidate the role of SNF3 (Sucrose Non Fermenting 3) as a glucose sensor and its possible role in contributing to the fitness and survivability of C. glabrata in glucose-limited environment. The SNF3 knockout strain was constructed and subjected to different glucose concentrations to evaluate its growth, biofilm formation, amphotericin B susceptibility, ex vivo survivability and effects on the transcriptional profiling of the sugar receptor repressor (SRR) pathway-related genes. The CgSNF3Δ strain showed a retarded growth in low glucose environments (0.01 and 0.1%) in both fermentation and respiration-preferred conditions but grew well in high glucose concentration environments (1 and 2%). It was also found to be more susceptible to amphotericin B in low glucose environment (0.1%) and macrophage engulfment but showed no difference in the biofilm formation capability. The deletion of SNF3 also resulted in the down-regulation of about half of hexose transporters genes (four out of nine). Overall, the deletion of SNF3 causes significant reduction in the ability of C. glabrata to sense limited surrounding glucose and consequently disrupts its competency to transport and perform the uptake of this critical nutrient. This study highlighted the role of SNF3 as a high affinity glucose sensor and its role in aiding the survivability of C. glabrata particularly in glucose limited environment.