At the interface of life and death : post-mortem and other applications of vaginal, skin, and salivary microbiome analysis in forensics

Microbial forensics represents a promising tool to strengthen traditional forensic investigative methods and fill related knowledge gaps. Large-scale microbiome studies indicate that microbial fingerprinting can assist forensics in areas such as trace evidence, source tracking, geolocation, and circumstances of death. Nevertheless, the majority of forensic microbiome studies focus on soil and internal organ samples, whereas the microbiome of skin, mouth, and especially vaginal samples that are routinely collected in sexual assault and femicide cases remain underexplored. This review discusses the current and emerging insights into vaginal, skin, and salivary microbiome-modulating factors during life (e.g., lifestyle and health status) and after death (e.g., environmental influences and post-mortem interval) based on next-generation sequencing. We specifically highlight the key aspects of female reproductive tract, skin, and mouth microbiome samples relevant in forensics. To fill the current knowledge gaps, future research should focus on the degree to which the post-mortem succession rate and profiles of vaginal, skin, and saliva microbiota are sensitive to abiotic and biotic factors, presence or absence of oxygen and other gases, and the nutrient richness of the environment. Application of this microbiome-related knowledge could provide valuable complementary data to strengthen forensic cases, for example, to shed light on the circumstances surrounding death with (post-mortem) microbial fingerprinting. Overall, this review synthesizes the present knowledge and aims to provide a framework to adequately comprehend the hurdles and potential application of vaginal, skin, and salivary post-mortem microbiomes in forensic investigations

Lactobacillus isalae sp. nov., isolated from the female reproductive tract

Abstract: A novel strain of the genus Lactobacillus, named AMBV1719T, was isolated from the vagina of a healthy participant in our largescale citizen science project on the female microbiome, named Isala. Phylogenetic analysis showed that the 16S rRNA gene of AMBV1719T is most similar to that of Lactobacillus taiwanensis with a sequence similarity of 99.873 %. However, a genome- wide comparison using average nucleotide identity (ANI) revealed that isolate AMBV1719T showed the highest ANI with Lactobacillus paragasseri JCM 5343T, with a value of only 88.17 %. This low ANI value with the most closely related strains known to date indicated that AMBV1719T represents a distinct species. This strain has a limited ability to degrade carbon sources compared to Lactobacillus gasseri, indicating its adaptation to the host. Its genome has a length of 2.12 Mb with a G+C content of 34.8 mol%. We thus propose the name Lactobacillus isalae sp. nov. for this novel species, with AMBV1719T (=LMG 32886T=CECT 30756T) as the type strain

Relationship between classic indicators of health behaviour and contraceptive choices in women in Flanders

Abstract: Background In this study we shed light on ongoing trends in contraceptive use in Flanders (Belgium). Building on the fundamental cause theory and social diffusion of innovation theory, we examine socio-economic gradients in contraceptive use and the relationship to health behaviours. Methods Using the unique and recently collected (2020) ISALA data, we used multinomial logistic regression to model the uptake of contraceptives and its association to educational level and health behaviour (N:4316 women). Results Higher educated women, and women with a healthy lifestyle especially, tend to use non-hormonal contraceptives or perceived lower-dosage hormonal contraceptives that are still trustworthy from a medical point of view. Moreover, we identified a potentially vulnerable group in terms of health as our results indicate that women who do not engage in preventive health behaviours are more likely to use no, or no modern, contraceptive method. Discussion The fact that higher educated women and women with a healthy lifestyle are less likely to use hormonal contraceptive methods is in line with patient empowerment, as women no longer necessarily follow recommendations by healthcare professionals, and there is a growing demand for naturalness in Western societies. Conclusion The results of this study can therefore be used to inform policy makers and reproductive healthcare professionals, since up-to-date understanding of women\u2019s contraceptive choices is clearly needed in order to develop effective strategies to prevent sexually transmitted infections and unplanned pregnancies, and in which women can take control over their sexuality and fertility in a comfortable and pleasurable way

Riboflavin for women's health and emerging microbiome strategies

Abstract: Riboflavin (vitamin B2) is an essential water-soluble vitamin that serves as a precursor of flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD). FMN and FAD are coenzymes involved in key enzymatic reactions in energy metabolism, biosynthesis, detoxification and electron scavenging pathways. Riboflavin deficiency is prevalent worldwide and impacts women's health due to riboflavin demands linked to urogenital and reproductive health, hormonal fluctuations during the menstrual cycle, pregnancy, and breastfeeding. Innovative functional foods and nutraceuticals are increasingly developed to meet women's riboflavin needs to supplement dietary sources. An emerging and particularly promising strategy is the administration of riboflavin-producing lactic acid bacteria, combining the health benefits of riboflavin with those of probiotics and in situ riboflavin production. Specific taxa of lactobacilli are of particular interest for women, because of the crucial role of Lactobacillus species in the vagina and the documented health effects of other Lactobacillaceae taxa in the gut and on the skin. In this narrative review, we synthesize the underlying molecular mechanisms and clinical benefits of riboflavin intake for women's health, and evaluate the synergistic potential of riboflavin-producing lactobacilli and other microbiota

Impacts of menstruation, community type, and an oral yeast probiotic on the vaginal microbiome

A healthy state of the vaginal microbiome can prevent vaginal disease and promote successful fertilization and healthy pregnancies. Little is known about the stability of the vaginal microbiome and the influence of factors such as diet and probiotics. While less explored, yeast probiotics have an interesting potential because of their immunomodulatory and pathogen inhibition capacities. In this study, we investigated the impact of the oral yeast probiotic Saccharomyces cerevisiae CNCM I-3856 on the vaginal microbiomes of 52 healthy women using 16S and internal transcribed spacer (ITS) amplicon sequencing and quantitative PCR (qPCR). The vaginal fungal loads remained low, even after oral yeast supplementation, complicating the analysis of the vaginal mycobiome. Lactobacillus crispatus and Lactobacillus iners were the most dominant species in our study population and were found to codominate in 23% of the baseline samples. Bifidobacterium, Streptococcus, and Prevotella were also frequently found. The microbiome profiles were dynamic: 69% of women showed a shift in the dominant community members at least once during the 42-day sampling period. In addition, lower Lactobacillus abundances were observed at the time points after menstruation. Higher relative abundances of Lactobacillus with more L. iners-dominated samples and a trend toward lower relative abundances of Prevotella were observed in the probiotic group, but analyses of the effects of the yeast probiotic were complicated by differences already present at the onset of the study. Thus, our findings especially highlighted that the impact of menstruation and the stratification of women based on the dominant vaginal taxa before randomization and inclusion is important for future research: while the impact of the yeast probiotic on vaginal microbiome in healthy women was limited. IMPORTANCE How to define and promote a healthy state of the vaginal microbiome is not well understood. Knowledge of which underlying factors shape the microbial community composition of the vagina and how to modulate them will contribute to vaginal disease prevention and improve fertility. Here, we found that taking the menstrual cycle into account when designing a microbiome study is highly recommended: menstruation also showed to be poses an interesting time point for intervention because of the drop in the abundance of L. crispatus. Furthermore, the early stratification of groups (e.g., placebo versus treatment) according to the dominant taxa can be of high added value since menstruation impacts vaginal taxa differently, i.e., L. iners remains stable, in contrast to L. crispatus

Salivary microbiome of healthy women of reproductive age

ABSTRACT The human salivary microbial community plays a crucial role in local and systemic diseases. Biological and lifestyle factors such as menstrual cycle, oral hygiene, and smoking have been documented to impact this community. However, while hormonal contraceptives are the most prescribed drug in healthy women and intimate partners play key roles in microbial exchange between humans, their impact on the salivary microbiome of women of reproductive age have been understudied. Additionally, the role of other lifestyle factors such as diet, allergies, age, and stress on the saliva microbiome of the general population is not well understood. Here, we studied the salivary microbiome of 255 healthy women of reproductive age using self-sampling kits and 16S rRNA amplicon sequencing combined with questionnaires on lifestyle and host-related parameters. A preserved salivary bacterial community of 12 genera (Actinobacillus, Actinomyces, Alloprevotella, Campylobacter, Fusobacterium, Gemella, Granulicatella, Leptotrichia, Neisseria, Prevotella, Streptococcus, and Veillonella) was identified. Contrary to what we expected, the number of intimate partners or specific contraceptive use did not have a major impact on these bacterial communities. However, recent use of oral antibiotics was associated with a significant decrease in richness at genus level and increase in mean relative abundances of several taxa. Being stressed or nervous was associated with a significantly increased richness of the salivary microbiome at the level of amplicon sequencing variants . Nevertheless, these associations with host-related and lifestyle variables only appeared to be subtle, suggesting that the salivary microbiome is mainly driven by the buccal environment and health status of an individual. IMPORTANCE The salivary microbiome has been proven to play a crucial role in local and systemic diseases. Moreover, the effects of biological and lifestyle factors such as oral hygiene and smoking on this microbial community have already been explored. However, what was not yet well understood was the natural variation of the saliva microbiome in healthy women and how this is associated with specific use of hormonal contraception and with the number of different sexual partners with whom microbiome exchange is expected regularly. In this paper, we characterized the salivary microbiome of 255 healthy women of reproductive age using an in-depth questionnaire and self-sampling kits. Using the large metadata set, we were able to investigate the associations of several host-related and lifestyle variables with the salivary microbiome profiles. Our study shows a high preservation between individuals

Spontaneous riboflavin-overproducing Limosilactobacillus reuteri for biofortification of fermented foods

Riboflavin-producing lactic acid bacteria represent a promising and cost-effective strategy for food biofortification, but production levels are typically insufficient to support daily human requirements. In this study, we describe the novel human isolate Limosilactobacillus reuteri AMBV339 as a strong food biofortification candidate. This strain shows a high natural riboflavin (vitamin B2) overproduction of 18.36 μg/ml, biomass production up to 6 × 10(10) colony-forming units/ml (in the typical range of model lactobacilli), and pH-lowering capacities to a pH as low as 4.03 in common plant-based (coconut, soy, and oat) and cow milk beverages when cultured up to 72 h at 37°C. These properties were especially pronounced in coconut beverage and butter milk fermentations, and were sustained in co-culture with the model starter Streptococcus thermophilus. Furthermore, L. reuteri AMBV339 grown in laboratory media or in a coconut beverage survived in gastric juice and in a simulated gastrointestinal dialysis model with colon phase (GIDM-colon system) inoculated with fecal material from a healthy volunteer. Passive transport of L. reuteri AMBV339-produced riboflavin occurred in the small intestinal and colon stage of the GIDM system, and active transport via intestinal epithelial Caco-2 monolayers was also demonstrated. L. reuteri AMBV339 did not cause fecal microbiome perturbations in the GIDM-colon system and inhibited enteric bacterial pathogens in vitro. Taken together, our data suggests that L. reuteri AMBV339 represents a promising candidate to provide riboflavin fortification of plant-based and dairy foods, and has a high application potential in the human gastrointestinal tract