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. 2020 Dec 9;8(12):1949.
doi: 10.3390/microorganisms8121949.

Distinct Functional Traits of Lactobacilli from Women with Asymptomatic Bacterial Vaginosis and Normal Microbiota

Rinku Pramanick  1 , Clara Aranha  1
Affiliations

Distinct Functional Traits of Lactobacilli from Women with Asymptomatic Bacterial Vaginosis and Normal Microbiota

Rinku Pramanick et al. Microorganisms. .

Abstract

Asymptomatic bacterial vaginosis (BV) in reproductive-age women has serious obstetric and gynecological consequences. Despite its high incidence, the behavior of vaginal lactobacilli in asymptomatic BV is unknown. We analyzed the functional properties of previously isolated vaginal lactobacilli from asymptomatic women with normal, intermediate, and BV microbiota. Lactic acid and antimicrobial activity against seven urogenital pathogens were evaluated from lactobacilli cell-free culture supernatants (CFCs) (n = 207) after 48 h incubation in MRS. Lactobacilli isolates were used to evaluate H2O2, autoaggregation and coaggregation with C. albicans. Lactobacilli from normal microbiota produced more d-lactate than lactobacilli from intermediate and asymptomatic BV (p = 0.007). L. plantarum, L. fermentum and L. reuteri produced greater d-lactate whereas L. rhamnosus, L. crispatus, L. johnsonii were greater producers of l-lactate. Interspecies positive correlation was observed in the lactic acid contents of CFCs. Distribution of H2O2-producing lactobacilli did not vary significantly among the groups. When lactic acid isomers were considered, species from intermediate and BV microbiota clustered together with each other and distinctly from species of normal microbiota. Broad-spectrum antagonism (≥90% inhibition) against E. coli, C. albicans, S. aureus, P. aeruginosa, G. vaginalis, N. gonorrhoeae, S. agalactiae were displayed by 46.86% (97) of isolates. Our study highlights the differential functional properties of vaginal lactobacilli from women with normal microbiota and asymptomatic BV.

Keywords: Lactobacillus; asymptomatic BV; bacterial vaginosis; lactic acid; probiotic properties; vaginal microbiota.

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Conflict of interest statement

The authors declare that there are no conflict of interest in the present study.

Figures

Figure 1
Figure 1
The acidity of CFCs (cell-free culture supernatants) was measured after the growth of different Lactobacillus species. The pH of the CFCs varied significantly among the species, as determined by the Kruskal–Wallis test. The topmost line indicates the Kruskal–Wallis test and downward pointing lines indicate Dunn’s multiple comparison test. * p < 0.05, ** p < 0.001, *** p < 0.001. Data are represented as a box plot wherein the box indicates the interquartile ranges, line within the bars represents the median and whiskers represent the minimum and the maximum values.
Figure 2
Figure 2
Lactic concentrations: (a) total lactic acid, (b) d-lactic acid, (c) l-lactic acid, (d) d-/l-lactic acid ratio of lactobacilli recovered from different vaginal microbiota. The amount of lactic acid isomers and d-/l-lactic acid ratio of lactobacilli metabolites differed significantly between the groups. Data are represented as mean ± SD. Ns = not significant and * p < 0.05, ** p < 0.01 indicating statistical significance after performing the Kruskal–Wallis test.
Figure 3
Figure 3
Lactic concentrations: (a) total lactic acid, (b) d-lactic acid, (c) l-lactic acid, (d) d-/l-lactic acid ratio of lactobacilli recovered from different vaginal microbiota. The amount of lactic acid isomers and d-/l-lactic acid ratio of lactobacilli metabolites differed significantly between the groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 indicating statistical significance after performing the Kruskal–Wallis test. The topmost line indicates the Kruskal–Wallis test and downward pointing lines indicate Dunn’s multiple comparison test. Data are represented as a box plot wherein the box indicates the interquartile ranges, line within the bars represents the median and whiskers represent the minimum and the maximum values.
Figure 4
Figure 4
d-/l- lactic ratios of (a) L. jensenii, (b) L. gasseri, (c) L. reuteri, (d) L. salivarius, (e) L. johnsonii, (f) L. rhamnosus from normal, intermediate and bacterial vaginosis (BV) microbiota. Data are represented as mean ± SD. ns =not significant and * p < 0.05, indicates statistical significance after performing the Kruskal–Wallis test.
Figure 5
Figure 5
Correlation between Lactobacillus species from normal microbiota in d-lactic acid production (a to d- top panel) (a) L. gasseri with L. reuteri, (b) L. reuteri with L. rhamnosus, (c) L. rhamnosus with L. crispatus, (d) L. fermentum with L. reuteri and l- lactic acid production (e to h- bottom panel) of (e) L. gasseri with L. reuteri, (f) L. reuteri with L. rhamnosus, (g) L. rhamnosus with L. crispatus, (h) L. fermentum with L. reuteri. Correlations between species were carried out by Spearman’s rank correlation. The correlation coefficient and p value for each analysis are mentioned in the graph.
Figure 6
Figure 6
Autoaggregation of (a) Lactobacillus isolates from different groups and (b) species-specific % autoaggregation. Candida coaggregation of (c) Lactobacillus from different microbiota and (d) coaggregation of various species. The topmost line indicates the Kruskal–Wallis test and downward pointing lines indicate Dunn’s multiple comparison test. Data are represented as a box plot wherein the box indicates the interquartile ranges, line within the bars represents the median and whiskers represent the minimum and the maximum values. ns = not significant, * p < 0.05, ** p < 0.01.
Figure 7
Figure 7
Lactobacilli with broad-spectrum antimicrobial activity. Lactobacilli isolates showing ≥90% growth inhibition on all the tested urogenital pathogens. (a) Proportions of lactobacilli isolates from normal, intermediate, and BV microbiota. The parts of the whole plot indicate that 76.29%, 8.25%, 15.46% of the isolates with broad-spectrum antimicrobial activity belonged to normal, intermediate and BV microbiota, respectively. (b) Lactobacillus species distribution from each normal, intermediate, and BV microbiota. The stack bar graph indicates the diversity of lactobacilli with broad-spectrum antimicrobial potential in normal microbiota is different from intermediate and BV microbiota.
Figure 8
Figure 8
Heatmap of concentrations of lactic acid produced by different species of Lactobacillus present in normal, intermediate and asymptomatic BV microbiota. Mean values of d- and l-lactic acid amount in CFCs of each species from the three groups were used for hierarchical clustering. Lactobacillus species are shown on the right side and the microbiota from which they were isolated are represented on the left side of the heatmap. A color bar with scales is shown, indicating that dark red corresponds to the maximum value and dark blue to minimum value.
Figure 9
Figure 9
Relationship of different functional properties of lactobacilli with each other. The Spearman’s correlation matrix indicates the correlation nature between the lactobacilli probiotic traits. Dark blue color on the scale denotes a perfect positive correlation (r = 1) and dark red indicates a perfect negative correlation (r = −1).
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References

    1. Wiesenfeld H.C., Hillier S.L., Krohn M.A., Landers D.V., Sweet R.L. Bacterial vaginosis is a strong predictor of Neisseria gonorrhoeaee and Chlamydia trachomatis infection. Clin. Infect. Dis. 2003;36:663–668. doi: 10.1086/367658. - DOI - PubMed
    1. Schwebke J.R. Abnormal vaginal flora as a biological risk factor for acquisitionof HIV infection and sexually transmitted diseases. J. Infect. Dis. 2005;192:1315–1317. doi: 10.1086/462430. - DOI - PubMed
    1. Brotman R.M. Vaginal microbiome and sexually transmitted infections: An epidemiologic perspective. J. Clin. Investig. 2011;121:4610–4617. doi: 10.1172/JCI57172. - DOI - PMC - PubMed
    1. Koumans E.H., Sternberg M., Bruce C., McQuillan G., Kendrick J., Sutton M., Markowitz L.E. The Prevalence of Bacterial Vaginosis in the United States, 2001–2004; Associations with Symptoms, Sexual Behaviors, and Reproductive Health. Sex. Transm. Dis. 2007;34:864–869. doi: 10.1097/OLQ.0b013e318074e565. - DOI - PubMed
    1. Joyisa N., Moodley D., Nkosi T., Talakgale R., Sebitloane M., Naidoo M., Karim Q.A. Asymptomatic Bacterial Vaginosis in Pregnancy and Missed Opportunities for Treatment: A Cross-Sectional Observational Study. Infect. Dis. Obstet. Gynecol. 2019;2019:7808179. doi: 10.1155/2019/7808179. - DOI - PMC - PubMed

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