Gardnerella Vaginalis Dominates Multi-Species Biofilms in both Pre-Conditioned and Competitive In Vitro Biofilm Formation Models

Roșca, A.; Castro, Joana; França, Ângela; Vaneechoutte, Mario; Cerca, Nuno

doi:10.1007/s00248-021-01917-2

Cited by 20 publications

(19 citation statements)

References 44 publications

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“…Despite BV has been associated with a polymicrobial biofilm, we first tested the EO antimicrobial activity against single-species biofilms, for comparative purposes. As shown before ( Rosca et al., 2021 ), the ability to form single-species biofilms varied significantly between species, with P. bivia and M. curtisii forming negligible biofilms after 24 h of incubation. Interestingly, when using the highest MLC concentration determined for the selected species (0.63 µL mL -1 ), most biofilms biomass was eradicated, except for G. vaginalis and L. iners ( Figure 1A ).…”

Section: Resultssupporting

confidence: 71%

“…Although BV is associated with the presence of a polymicrobial biofilm on the vaginal epithelium ( Swidsinski et al., 2005 ), up to date, no functional in vitro studies have assessed BV-associated biofilms composed of more than three different species ( Rosca et al., 2021 ). Since in vivo evidence supports the role of more species to be present during BV, in this study, we aimed to assess if six relevant BV-associated species could, in fact, establish a polymicrobial biofilm.…”

Section: Resultsmentioning

confidence: 99%

“…Gardnerella vaginalis ATCC 14018 and other five cultivable bacterial species associated with BV were used in the current study, namely Fannyhessea vaginae ATCC BAA-55, Lactobacillus iners CCUG 28746, Mobiluncus curtisii ATCC 35241, Peptostreptococcus anaerobius ATCC 27337, and Prevotella bivia ATCC 29303. All species were grown as described before ( Rosca et al., 2021 ), on plates containing Columbia Blood Agar Base medium (CBA) (Oxoid, Basingstoke, UK) supplemented with 5% (v/v) defibrinated horse blood (Oxoid) and incubated, for 48 h, at 37°C in the presence of 10% carbon dioxide ( G. vaginalis ) or in anaerobic conditions (the other five BV-associated species) generated by using AnaeroGen sachets (Thermo Fisher Scientific, Gloucester, UK) in sealed jars (Oxoid).…”

Section: Methodsmentioning

confidence: 99%

“…A previously described in vitro polymicrobial biofilm formation model ( Rosca et al., 2021 ) was followed in this study, but using six, instead of three BV-associated species. Briefly, the inoculum of each species was grown in NYC III broth and incubated for 24 h at 37°C under anaerobic conditions, as above mentioned.…”

Section: Methodsmentioning

confidence: 99%

“…Subsequently, the 24 h cultures were adjusted to a concentration of ~ 1.0 × 10 7 cells mL -1 , and the six species were further co-incubated anaerobically in 24-well tissue culture plates (Orange Scientific) for 24 h at 37°C. We used ~1.0 × 10 7 cells mL -1 as starting inoculum for each species, since we have previously demonstrated it to be a good starting concentration, to allow bacterial integration in the polymicrobial biofilms ( Castro et al., 2021 ; Rosca et al., 2021 ). The final volume in each well was 1 mL.…”

Section: Methodsmentioning

confidence: 99%

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Six Bacterial Vaginosis-Associated Species Can Form an In Vitro and Ex Vivo Polymicrobial Biofilm That Is Susceptible to Thymbra capitata Essential Oil

Roșca

Castro

Sousa

et al. 2022

Front. Cell. Infect. Microbiol.

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Bacterial vaginosis (BV) is associated with serious gynaecologic and obstetric complications. The hallmark of BV is the presence of a polymicrobial biofilm on the vaginal epithelium, but BV aetiology is still a matter of debate. We have previously developed an in vitro biofilm model that included three BV-associated species, but, up to now, no studies are available whereby more bacterial species are grown together to better mimic the in vivo situation. Herein, we characterized the first polymicrobial BV biofilm consisting of six cultivable BV-associated species by using both in vitro and ex vivo vaginal tissue models. Both models revealed that the six species were able to incorporate the polymicrobial biofilm, at different bacterial concentrations. As it has been thought that this polymicrobial biofilm may increase the survival of BV-associated species when exposed to antibiotics, we also assessed if the Thymbra capitata essential oil (EO), which has recently been shown to be highly bactericidal against several Gardnerella species, could maintain its anti-biofilm activity against this polymicrobial biofilm. Under our experimental conditions, T. capitata EO exhibited a high antibacterial effect against polymicrobial biofilms, in both tested models, with a significant reduction in the biofilm biomass and the number of culturable cells. Overall, this study shows that six BV-associated species can grow together and form a biofilm both in vitro and when using an ex vivo model. Moreover, the data obtained herein should be considered in further applications of T. capitata EO as an antimicrobial agent fighting BV.

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Section: Resultssupporting

confidence: 71%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Six Bacterial Vaginosis-Associated Species Can Form an In Vitro and Ex Vivo Polymicrobial Biofilm That Is Susceptible to Thymbra capitata Essential Oil

Roșca

Castro

Sousa

et al. 2022

Front. Cell. Infect. Microbiol.

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Efficacy of Dequalinium Chloride vs Metronidazole for the Treatment of Bacterial Vaginosis

Raba,

Ďurkech,

Malík

et al. 2024

JAMA Netw Open

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ImportanceBacterial vaginosis (BV) is a common cause of vaginal infection. First-line treatments of BV are metronidazole and clindamycin. Due to the increase in antibiotic resistance, effective nonantibiotic treatments for BV are needed.ObjectiveTo examine whether dequalinium chloride, a broad-spectrum antiseptic, is noninferior to oral metronidazole for the treatment of BV.Design, Setting, and ParticipantsThis phase 4, multicenter, triple-blind, double-dummy, parallel, noninferiority randomized clinical trial was conducted from July 29, 2021, to August 25, 2022, with a 1-month follow-up. Participants were premenopausal women 18 years or older with BV from 11 gynecologic practices and 1 hospital in Poland, Slovakia, and the Czech.InterventionPatients were randomized to treatment with dequalinium chloride vaginal tablets (10 mg once daily for 6 days) or oral metronidazole (500 mg twice daily for 7 days). Double-dummy medication kits contained vaginal and oral tablets with placebo and active medication.Main Outcomes and MeasuresThe main outcome was the noninferiority margin (of 15 percentage points) in the absolute difference in clinical cure rates between dequalinium chloride and metronidazole 7 to 11 days after start of treatment (visit 1). Noninferiority was met if the lower 95% CI for the difference in clinical cure rate was less than 15 percentage points at visit 1.ResultsA total of 147 women (mean [SD] age, 36.7 [9.0] years) were treated with dequalinium chloride (n = 72) or metronidazole (n = 75). The clinical cure rates at visit 1 were 64 of 69 (92.8%) for dequalinium chloride vs 69 of 74 (93.2%) for metronidazole in the intention-to-treat population, whereas in the per-protocol population, cure rates were 54 of 58 (93.1%) for dequalinium chloride vs 48 of 53 (90.6%) for metronidazole. The treatment differences of −0.5 percentage points (95% CI, −10.8 to 9.8 percentage points; P = .002) in the intention-to-treat population and 2.5 percentage points (95% CI, −9.4 to 14.4 percentage points; P = .001) in the per-protocol population confirmed the noninferiority of dequalinium chloride. The tolerability of dequalinium chloride was rated as very good by 30 of 50 patients (60.0%) but only by 21 of 54 (38.9%) for metronidazole. Three patients in the metronidazole group suspended treatment due to an adverse event.Conclusions and RelevanceThis randomized clinical trial showed that dequalinium chloride was not inferior to metronidazole for the treatment of BV. Dequalinium chloride had a similarly high cure rate but with better tolerability and fewer adverse events. With a similar efficacy to metronidazole and clindamycin, dequalinium chloride warrants consideration as first-line treatment for BV to help reduce antibiotic consumption.Trial RegistrationEudraCT: 2020-002489-15

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Gardnerella vaginalis, Fannyhessea vaginae, and Prevotella bivia Strongly Influence Each Other's Transcriptome in Triple-Species Biofilms

Sousa,

Novak,

França

et al. 2024

Microb Ecol

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Bacterial vaginosis (BV), the most common vaginal infection worldwide, is characterized by the development of a polymicrobial biofilm on the vaginal epithelium. While Gardnerella spp. have been shown to have a prominent role in BV, little is known regarding how other species can influence BV development. Thus, we aimed to study the transcriptome of Gardnerella vaginalis, Fannyhessea vaginae, and Prevotella bivia, when growing in triple-species biofilms. Single and triple-species biofilms were formed in vitro, and RNA was extracted and sent for sequencing. cDNA libraries were prepared and sequenced. Quantitative PCR analysis (qPCR) was performed on the triple-species biofilms to evaluate the biofilm composition. The qPCR results revealed that the triple-species biofilms were mainly composed by G. vaginalis and P. bivia was the species with the lowest percentage. The RNA-sequencing analysis revealed a total of 432, 126, and 39 differentially expressed genes for G. vaginalis, F. vaginae, and P. bivia, respectively, when growing together. Gene ontology enrichment of G. vaginalis downregulated genes revealed several functions associated with metabolism, indicating a low metabolic activity of G. vaginalis when growing in polymicrobial biofilms. This work highlighted that the presence of 3 different BV-associated bacteria in the biofilm influenced each other’s transcriptome and provided insight into the molecular mechanisms that enhanced the virulence potential of polymicrobial consortia. These findings will contribute to understand the development of incident BV and the interactions occurring within the biofilm.

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Gardnerella Vaginalis Dominates Multi-Species Biofilms in both Pre-Conditioned and Competitive In Vitro Biofilm Formation Models

Cited by 20 publications

References 44 publications

Six Bacterial Vaginosis-Associated Species Can Form an In Vitro and Ex Vivo Polymicrobial Biofilm That Is Susceptible to Thymbra capitata Essential Oil

Six Bacterial Vaginosis-Associated Species Can Form an In Vitro and Ex Vivo Polymicrobial Biofilm That Is Susceptible to Thymbra capitata Essential Oil

Efficacy of Dequalinium Chloride vs Metronidazole for the Treatment of Bacterial Vaginosis

Gardnerella vaginalis, Fannyhessea vaginae, and Prevotella bivia Strongly Influence Each Other's Transcriptome in Triple-Species Biofilms

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