Biology subjects

Wittouck, S.

Publications and source records attributed to Wittouck, S..

7 recordsLinked to original sources

bioRxiv2026

Exopolysaccharides of Lactobacillus crispatus mediate key balancing interactions with the vaginal mucosa

Lactobacillus crispatus is a dominant member of the healthy vaginal microbiota, yet the mechanisms by which it modulates host immunity remain poorly defined, in part due to the lack of tractable in vivo models. Here, we integrate bacterial genetics, in vitro epithelial systems, human-derived data and proteomic approach (Olink) to uncover a critical role for L. crispatus exopolysaccharides (EPS) in shaping the bacteria-vagina interactions. Comparative genomics identified a conserved EPS biosynthetic locus, with the priming glycosyltransferase gene epsE emerging as a regulatory node, in line with its distinct expression in human vaginal samples. Functional disruption of epsE abrogated L. crispatus EPS production and revealed its role for immune modulation. In human vaginal epithelial monolayers, EPS presence enhanced immune-regulatory (LAP TGF-beta-1) and anti-inflammatory (CST5) responses, whereas its absence triggered elevated pro-inflammatory cytokines (IL-1{beta}, IL-6, IL-8) and matrix metalloproteinase (MMP-10). In a 3D vaginal organotypic model, EPS increased chemokines (CXCL5, CXCL6) linked to immune surveillance and the presence of the markers was validated in vaginal samples of healthy volunteers. These findings position EPS as a key immunomodulatory structure of L. crispatus, advancing our mechanistic understanding of host-commensal interactions and informing microbiome-based strategies to promote vaginal health.

microbiology↗

bioRxiv2025

From Diversity to Dominance: How Salt and CO2 Shape LAB-dominated Ecosystems in Vegetable Fermentations

Research on microbial ecosystems is often challenging due to high diversity of microbial taxa present and the complexity of controlling environmental variables. To address these challenges, fermented foods are simpler and more reproducible model ecosystems, where both community composition and environmental factors can be more precisely controlled and manipulated. In this study, we focused on fermented vegetables which are typically dominated by lactic acid bacteria (LAB). It is not completely understood why lactic acid bacteria (LAB) consistently drive the spontaneous fermentation of vegetables such as cabbage and carrots and how variables such as vegetable substrates, salt addition, and carbon dioxide levels can impact microbial community dynamics. Here, we explored the temporal microbial dynamics in standardized fermentations of 11 different vegetables (including beetroot, bell pepper, cabbage, carrot, cucumber, fennel, green asparagus, leek, parsnip, sunroot, and tomato), revealing a consistent dominance of Leuconostoc and other LAB. Additionally, we investigated the impact of varying salt concentrations, demonstrating that lower salt levels resulted in a delayed appearance of the typically dominant LAB community, while simultaneously revealing a higher abundance of Weissella and various Enterobacterales taxa. These effects imposed by reduced salt concentrations were mitigated by CO2 injection, which reverted the enhanced Enterobacterales levels and increased the overall abundance of Lactobacillales. This study demonstrates how targeted manipulation of environmental parameters, such as salinity and gas composition, can be used to uncover ecological principles governing microbial succession and community assembly in reproducible fermentation-based model ecosystems. ImportanceUnderstanding the ecological principles that shape microbial community assembly is essential for advancing our knowledge of microbial ecosystems. Fermented vegetables, increasingly popular among the general population, provide tractable and reproducible model systems to study microbial succession under controlled environmental conditions. By systematically manipulating variables such as vegetable type, salinity and gas composition, we uncovered the effect of these factors on the microbial dynamics throughout the fermentation. These insights not only contribute to a better understanding of the microbial ecology of these man-made food systems but also suggest directions for novel strategies to optimize fermentation processes for producing faster, safer, and more flavorful foods.

microbiology↗

bioRxiv2025

First-void urine and cervicovaginal brushes as sampling devices for vaginal microbiome studies: a proof-of-concept study

This proof-of-concept study compares first-void urine (FVU) and cervicovaginal brushes to vaginal swabs for microbiome profiling in ten women. FVU and brushes provided comparable results in most cases, though storage conditions influenced microbial composition, especially for FVU. Despite limitations in sample size and diversity in participants, findings support these methods potential for self-sampling in womens health research, emphasizing the need for optimized techniques to improve vaginal microbiome profiling.

microbiology↗

bioRxiv2025

The female intimate microbiome space

Despite its importance in health, the female skin microbiome remains understudied. In this study we explored microbial dispersal across intimate body sites, including the vagina, groins, breast and mouth. Microbial similarity correlated with physical proximity, suggesting dispersal influenced by hygiene or sexual activity. Notably, lactobacilli were unexpectedly abundant on breast skin. These findings highlight the need for research into microbiome dynamics and their implications for womens health.

microbiology↗

bioRxiv2019

Comparative genome analysis of Lactobacillus mudanjiangensis, an understudied member of the Lactobacillus plantarum group

The genus Lactobacillus is known to be extremely diverse and consists of different phylogenetic groups that show a diversity roughly equal to the expected diversity of a typical bacterial genus. One of the most prominent phylogenetic groups within this genus is the Lactobacillus plantarum group which contains the understudied Lactobacillus mudanjiangensis species. Before this study, only one L. mudanjiangensis strain, DSM 28402T, was described but without whole-genome analysis. In this study, three strains classified as L. mudanjiangensis, were isolated from three different carrot juice fermentations and their whole-genome sequence was determined, together with the genome sequence of the type strain. The genomes of all four strains were compared with publicly available L. plantarum group genome sequences. This analysis showed that L. mudanjiangensis harbored the second largest genome size and gene count of the whole L. plantarum group. In addition, all members of this species showed the presence of a gene coding for a putative cellulose-degrading enzyme. Finally, three of the four L. mudanjiangensis strains studied showed the presence of pili on scanning electron microscopy (SEM) images, which were linked to conjugative gene regions, coded on plasmids in at least two of the strains studied. Author summaryLactobacillus mudanjiangensis is an understudied species within the Lactobacillus plantarum group. Since its first description, no other studies have reported its isolation. Here, we present the first four genome sequences of this species, which include the genome sequence of the type strain and three new L. mudanjiangensis strains isolated from fermented carrot juice. The genomes of all four strains were compared with publicly available L. plantarum group genome sequences. We found that this species harbored the second largest genome size and gene count of the whole L. plantarum group. Furthermore, we present the first scanning electron microscopy (SEM) images of L. mudanjiangensis, which showed the formation of pili in three strains that we linked to genes related to conjugation. Finally, we found the presence of a unique putative cellulose-degrading enzyme, opening the door for different industrial applications of these Lactobacillus strains.

microbiology↗

bioRxiv2019

A genome-based species taxonomy of the Lactobacillus Genus Complex

BackgroundThere are over 200 published species within the Lactobacillus Genus Complex (LGC), the majority of which have sequenced type strain genomes available. Although gold standard, genome-based species delimitation cutoffs are accepted by the community, they are seldom checked against currently available genome data. In addition, there are many species-level misclassification issues within the LGC. We constructed a de novo species taxonomy for the LGC based on 2,459 publicly available, decent-quality genomes and using a 94% core nucleotide identity threshold. We reconciled these de novo species with published species and subspecies names by (i) identifying genomes of type strains in our dataset and (ii) performing comparisons based on 16S rRNA sequence identity against type strains. ResultsWe found that genomes within the LGC could be divided into 239 clusters (de novo species) that were discontinuous and exclusive. Comparison of these de novo species to published species lead to the identification of ten sets of published species that can be merged and one species that can be split. Further, we found at least eight genome clusters that constitute new species. Finally, we were able to accurately classify 98 unclassified genomes and reclassify 74 wrongly classified genomes. ConclusionsThe current state of LGC species taxonomy is largely consistent with genome data, but there are some inconsistencies as well as genome misclassifications. These inconsistencies should be resolved to evolve towards a meaningful taxonomy where species have a consistent size in terms of sequence divergence.

bioinformatics↗

bioRxiv2018

Topical cream with live lactobacilli modulates the skin microbiome and reduce acne symptoms

The skin is home to an important part of our commensal microbiota, despite it being a cool, acidic and desiccated environment. Tailored microbiome modulation approaches with, for example probiotics, are highly challenging for this body site. Here we show by next-generating sequencing that Lactobacillus taxa -especially those known to be dominant in the human vagina- are underestimated members of the skin microbiota. Specific Lactobacillus strains were selected in the lab and formulated in a viable form in an oil in water-based topical cream. Facial application by patients with mild-to-moderate acne symptoms was able to reduce inflammatory lesions and comedone formation. This was associated with a temporary modulation of the skin microbiome, including a reduction in relative abundance of staphylococci and an increase in lactobacilli. Skin microbiome modulation by addition of carefully formulated lactobacilli seems to be new therapeutic option to reduce antibiotic use for common acne symptoms.

clinical trials↗