scholarly journals Atopobium vaginae and Prevotella bivia Are Able to Incorporate and Influence Gene Expression in a Pre-Formed Gardnerella vaginalis Biofilm

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 247
Author(s):  
Joana Castro ◽  
Aliona S. Rosca ◽  
Christina A. Muzny ◽  
Nuno Cerca
Keyword(s):  
Virulence Genes ◽  
Gardnerella Vaginalis ◽  
Bacterial Populations ◽  
Biofilm Model ◽  
Prevotella Bivia ◽  
The Impact ◽  
Influence Gene Expression ◽  
Pna Fish ◽  
Atopobium Vaginae

Bacterial vaginosis (BV) is associated with a highly structured polymicrobial biofilm on the vaginal epithelium where Gardnerella species presumably play a pivotal role. Gardnerella vaginalis, Atopobium vaginae, and Prevotella bivia are vaginal pathogens detected during the early stages of incident BV. Herein, we aimed to analyze the impact of A. vaginae and P. bivia on a pre-established G. vaginalis biofilm using a novel in vitro triple-species biofilm model. Total biofilm biomass was determined by the crystal violet method. We also discriminated the bacterial populations in the biofilm and in its planktonic fraction by using PNA FISH. We further analyzed the influence of A. vaginae and P. bivia on the expression of key virulence genes of G. vaginalis by quantitative PCR. In our tested conditions, A. vaginae and P. bivia were able to incorporate into pre-established G. vaginalis biofilms but did not induce an increase in total biofilm biomass, when compared with 48-h G. vaginalis biofilms. However, they were able to significantly influence the expression of HMPREF0424_0821, a gene suggested to be associated with biofilm maintenance in G. vaginalis. This study suggests that microbial relationships between co-infecting bacteria can deeply affect the G. vaginalis biofilm, a crucial marker of BV.

scholarly journals Prospective Evaluation of Molecular Assays for Diagnosis of Vaginitis

2019 ◽  
Vol 58 (1) ◽  
Author(s):  
Sandra S. Richter ◽  
Joshua Otiso ◽  
Oluwatosin J. Goje ◽  
Sherilynn Vogel ◽  
Jory Aebly ◽  
...  
Keyword(s):  
Gardnerella Vaginalis ◽  
Reference Standard ◽  
Content Type ◽  
Molecular Assays ◽  
Molecular Tests ◽  
Normal Microbiota ◽  
The Impact ◽  
Specific Reagent ◽  
Atopobium Vaginae

ABSTRACT Molecular tests to diagnose conditions involving the disruption of normal microbiota are difficult to optimize. Using Nugent-scored Gram stain (NS) as the reference standard, we evaluated the performance of 3 molecular assays for the diagnosis of bacterial vaginosis (BV) and examined the impact of an incremental increase in bacterial targets. The BD Affirm assay includes a DNA probe for Gardnerella vaginalis, the Hologic transcription-mediated amplification (TMA) analyte-specific reagent (ASR) assay adds a second Lactobacillus sp. target, and the recently cleared in vitro diagnostic use (IVD) Aptima BV assay includes a third target (Atopobium vaginae). The diagnosis of vulvovaginal candidiasis (VVC) by the Affirm and Candida vaginitis Hologic TMA ASR assays was assessed using microscopy for yeast as the reference standard. From May to December 2018, 111 women with vaginitis symptoms prompting the clinician to order an Affirm test were enrolled with informed consent for the collection of additional specimens. Clinicians accurately predicted BV as the most likely diagnosis for 71% of the 45 patients with BV. Coinfection occurred in 13.5% of patients. For BV, the specificity of the Aptima IVD assay (86.3%) was higher than the Affirm assay (60.6%, P = 0.0002), but sensitivities were not significantly different. For VVC, the sensitivity of the ASR assay (100%) was higher than Affirm (75.9%; P = 0.023) and the specificity of the Affirm assay (98.8%) was higher than the ASR assay (86.6%; P = 0.004).

scholarly journals Relevance of Biofilm Models in Periodontal Research: From Static to Dynamic Systems

2021 ◽  
Vol 9 (2) ◽  
pp. 428
Author(s):  
María Carmen Sánchez ◽  
Andrea Alonso-Español ◽  
Honorato Ribeiro-Vidal ◽  
Bettina Alonso ◽  
David Herrera ◽  
...  
Keyword(s):  
Research Group ◽  
Surface Composition ◽  
Bacterial Colonization ◽  
Implant Surface ◽  
Biofilm Growth ◽  
Biofilm Model ◽  
Dental Implant Surface ◽  
The Impact

Microbial biofilm modeling has improved in sophistication and scope, although only a limited number of standardized protocols are available. This review presents an example of a biofilm model, along with its evolution and application in studying periodontal and peri-implant diseases. In 2011, the ETEP (Etiology and Therapy of Periodontal and Peri-Implant Diseases) research group at the University Complutense of Madrid developed an in vitro biofilm static model using representative bacteria from the subgingival microbiota, demonstrating a pattern of bacterial colonization and maturation similar to in vivo subgingival biofilms. When the model and its methodology were standardized, the ETEP research group employed the validated in vitro biofilm model for testing in different applications. The evolution of this model is described in this manuscript, from the mere observation of biofilm growth and maturation on static models on hydroxyapatite or titanium discs, to the evaluation of the impact of dental implant surface composition and micro-structure using the dynamic biofilm model. This evolution was based on reproducing the ideal microenvironmental conditions for bacterial growth within a bioreactor and reaching the target surfaces using the fluid dynamics mimicking the salivary flow. The development of this relevant biofilm model has become a powerful tool to study the essential processes that regulate the formation and maturation of these important microbial communities, as well as their behavior when exposed to different antimicrobial compounds.

scholarly journals Gardnerella vaginalis Enhances Atopobium vaginae Viability in an in vitro Model

2020 ◽  
Vol 10 ◽  
Author(s):  
Joana Castro ◽  
Aliona S. Rosca ◽  
Piet Cools ◽  
Mario Vaneechoutte ◽  
Nuno Cerca
Keyword(s):  
In Vitro Model ◽  
Gardnerella Vaginalis ◽  
Vitro Model ◽  
Atopobium Vaginae

scholarly journals Implications of Antimicrobial Combinations in Complex Wound Biofilms Containing Fungi

2017 ◽  
Vol 61 (9) ◽  
Author(s):  
Eleanor M. Townsend ◽  
Leighann Sherry ◽  
Ryan Kean ◽  
Donald Hansom ◽  
William G. Mackay ◽  
...  
Keyword(s):  
Foot Ulcer ◽  
Diabetic Foot Ulcer ◽  
Biofilm Model ◽  
Discernible Effect ◽  
Complex Wound ◽  
Bacteria And Fungi ◽  
Wound Biofilms ◽  
The Impact ◽  
Antimicrobial Combinations

ABSTRACT Diabetic foot ulcer treatment currently focuses on targeting bacterial biofilms, while dismissing fungi. To investigate this, we used an in vitro biofilm model containing bacteria and fungi, reflective of the wound environment, to test the impact of antimicrobials. Here we showed that while monotreatment approaches influenced biofilm composition, this had no discernible effect on overall quantity. Only by combining bacterium- and fungus-specific antibiotics were we able to decrease the biofilm bioburden, irrespective of composition.

scholarly journals Vaginal Microbiota of the Sexually Transmitted Infections Caused by Chlamydia trachomatis and Trichomonas vaginalis in Women with Vaginitis in Taiwan

2021 ◽  
Vol 9 (9) ◽  
pp. 1864
Author(s):  
Shu-Fang Chiu ◽  
Po-Jung Huang ◽  
Wei-Hung Cheng ◽  
Ching-Yun Huang ◽  
Lichieh Julie Chu ◽  
...  
Keyword(s):  
Chlamydia Trachomatis ◽  
Sexually Transmitted Infections ◽  
Trichomonas Vaginalis ◽  
Vaginal Microbiota ◽  
Gardnerella Vaginalis ◽  
Associated Bacteria ◽  
Sexually Transmitted ◽  
Prevotella Bivia ◽  
Initial Interaction

The three most common sexually transmitted infections (STIs) are Chlamydia trachomatis (CT), Neisseria gonorrhoeae (GC) and Trichomonas vaginalis (TV). The prevalence of these STIs in Taiwan remains largely unknown and the risk of STI acquisition affected by the vaginal microbiota is also elusive. In this study, a total of 327 vaginal swabs collected from women with vaginitis were analyzed to determine the presence of STIs and the associated microorganisms by using the BD Max CT/GC/TV molecular assay, microbial cultures, and 16S rRNA sequencing. The prevalence of CT, TV, and GC was 10.8%, 2.2% and 0.6%, respectively. A culture-dependent method identified that Escherichia coli and Streptococcus agalactiae (GBS) were more likely to be associated with CT and TV infections. In CT-positive patients, the vaginal microbiota was dominated by L. iners, and the relative abundance of Gardnerella vaginalis (12.46%) was also higher than that in TV-positive patients and the non-STIs group. However, Lactobacillus spp. was significantly lower in TV-positive patients, while GBS (10.11%), Prevotella bivia (6.19%), Sneathia sanguinegens (12.75%), and Gemella asaccharolytica (5.31%) were significantly enriched. Using an in vitro co-culture assay, we demonstrated that the growth of L. iners was suppressed in the initial interaction with TV, but it may adapt and survive after longer exposure to TV. Additionally, it is noteworthy that TV was able to promote GBS growth. Our study highlights the vaginal microbiota composition associated with the common STIs and the crosstalk between TV and the associated bacteria, paving the way for future development of health interventions targeting the specific vaginal bacterial taxa to reduce the risk of common STIs.

scholarly journals Crystal Violet Staining Alone Is Not Adequate to Assess Synergism or Antagonism in Multi-Species Biofilms of Bacteria Associated With Bacterial Vaginosis

2022 ◽  
Vol 11 ◽  
Author(s):  
Joana Castro ◽  
Ângela Lima ◽  
Lúcia G. V. Sousa ◽  
Aliona S. Rosca ◽  
Christina A. Muzny ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
Crystal Violet ◽  
Single Species ◽  
Gardnerella Vaginalis ◽  
Individual Species ◽  
Vaginal Epithelial Cells ◽  
Prevotella Bivia ◽  
Depth Study ◽  
Biofilm Quantification

Bacterial Vaginosis (BV) involves the presence of a multi-species biofilm adhered to vaginal epithelial cells, but its in-depth study has been limited due to the complexity of the bacterial community, which makes the design of in vitro models challenging. Perhaps the most common experimental technique to quantify biofilms is the crystal violet (CV) staining method. Despite its widespread utilization, the CV method is not without flaws. While biofilm CV quantification within the same strain in different conditions is normally accepted, assessing multi-species biofilms formation by CV staining might provide significant bias. For BV research, determining possible synergism or antagonism between species is a fundamental step for assessing the roles of individual species in BV development. Herein, we provide our perspective on how CV fails to properly quantify an in vitro triple-species biofilm composed of Gardnerella vaginalis, Fannyhessea (Atopobium) vaginae, and Prevotella bivia, three common BV-associated bacteria thought to play key roles in incident BV pathogenesis. We compared the CV method with total colony forming units (CFU) and fluorescence microscopy cell count methods. Not surprisingly, when comparing single-species biofilms, the relationship between biofilm biomass, total number of cells, and total cultivable cells was very different between each tested method, and also varied with the time of incubation. Thus, despite its wide utilization for single-species biofilm quantification, the CV method should not be considered for accurate quantification of multi-species biofilms in BV pathogenesis research.

scholarly journals Adhesion of Human ProbioticLactobacillus rhamnosusto Cervical and Vaginal Cells and Interaction with Vaginosis-Associated Pathogens

2008 ◽  
Vol 2008 ◽  
pp. 1-5 ◽  
Author(s):  
Sophie Coudeyras ◽  
Gwendoline Jugie ◽  
Marion Vermerie ◽  
Christiane Forestier
Keyword(s):  
Candida Albicans ◽  
Lactobacillus Rhamnosus ◽  
Probiotic Strain ◽  
Gardnerella Vaginalis ◽  
Promising Candidate ◽  
Division Rate ◽  
Adhesion Ability ◽  
Cell Division Rate ◽  
Prevotella Bivia

Objectives. The ability of a probioticLactobacillus rhamnosusstrain (Lcr35) to adhere to cervical and vaginal cells and to affect the viability of two main vaginosis-associated pathogens,Prevotella bivia, Gardnerella vaginalis, as well asCandida albicanswas investigated.Methods. Adhesion ability was determined in vitro with immortalized epithelial cells from the endocervix, ectocervix, and vagina. Coculture experiments were performed to count viable pathogens cells in the presence of Lcr35.Results. Lcr35 was able to specifically and rapidly adhere to the three cell lines. In coculture assays, a decrease in pathogen cell division rate was observed as from 4 hours of incubation and bactericidal activity after a longer period of incubation, mostly withP. bivia.Conclusion. The ability of Lcr35 to adhere to cervicovaginal cells and its antagonist activities against vaginosis-associated pathogens suggest that this probiotic strain is a promising candidate for use in therapy.

scholarly journals Adaptation of Staphylococcus aureus in a Medium Mimicking a Diabetic Foot Environment

Toxins ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 230
Author(s):  
Cassandra Pouget ◽  
Claude-Alexandre Gustave ◽  
Christelle Ngba-Essebe ◽  
Frédéric Laurent ◽  
Emmanuel Lemichez ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Diabetic Foot ◽  
In Vitro Model ◽  
Virulence Genes ◽  
Chronic Wounds ◽  
Ring Test ◽  
Vitro Model ◽  
The Impact

Staphylococcus aureus is the most prevalent pathogen isolated from diabetic foot infections (DFIs). The purpose of this study was to evaluate its behavior in an in vitro model mimicking the conditions encountered in DFI. Four clinical S. aureus strains were cultivated for 16 weeks in a specific environment based on the wound-like medium biofilm model. The adaptation of isolates was evaluated as follows: by Caenorhabditis elegans model (to evaluate virulence); by quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) (to evaluate expression of the main virulence genes); and by Biofilm Ring test® (to assess the biofilm formation). After 16 weeks, the four S. aureus had adapted their metabolism, with the development of small colony variants and the loss of b-hemolysin expression. The in vivo nematode model suggested a decrease of virulence, confirmed by qRT-PCRs, showing a significant decrease of expression of the main staphylococcal virulence genes tested, notably the toxin-encoding genes. An increased expression of genes involved in adhesion and biofilm was noted. Our data based on an in vitro model confirm the impact of environment on the adaptation switch of S. aureus to prolonged stress environmental conditions. These results contribute to explore and characterize the virulence of S. aureus in chronic wounds.

scholarly journals Acid Production by Vaginal Flora In Vitro Is Consistent with the Rate and Extent of Vaginal Acidification

1999 ◽  
Vol 67 (10) ◽  
pp. 5170-5175 ◽  
Author(s):  
E. R. Boskey ◽  
K. M. Telsch ◽  
K. J. Whaley ◽  
T. R. Moench ◽  
R. A. Cone
Keyword(s):  
Epithelial Cells ◽  
Growth Medium ◽  
Acid Production ◽  
Vaginal Flora ◽  
Gardnerella Vaginalis ◽  
Primary Role ◽  
Vaginal Ph ◽  
Prevotella Bivia ◽  
Human Vagina

Perinatally, and between menarche and menopause, increased levels of estrogen cause large amounts of glycogen to be deposited in the vaginal epithelium. During these times, the anaerobic metabolism of the glycogen, by the epithelial cells themselves and/or by vaginal flora, causes the vagina to become acidic (pH ∼4). This study was designed to test whether the characteristics of acid production by vaginal flora in vitro can account for vaginal acidity. Eight vaginalLactobacillus isolates from four species—L. gasseri, L. vaginalis, L. crispatus, andL. jensenii—acidified their growth medium to an asymptotic pH (3.2 to 4.8) that matches the range seen in theLactobacillus-dominated human vagina (pH 3.6 to 4.5 in most women) (B. Andersch, L. Forssman, K. Lincoln, and P. Torstensson, Gynecol. Obstet. Investig. 21:19–25, 1986; L. Cohen, Br. J. Vener. Dis. 45:241–246, 1969; J. Paavonen, Scand. J. Infect. Dis. Suppl. 40:31–35, 1983; C. Tevi-Bénissan, L. Bélec, M. Lévy, V. Schneider-Fauveau, A. Si Mohamed, M.-C. Hallouin, M. Matta, and G. Grésenguet, Clin. Diagn. Lab. Immunol. 4:367–374, 1997). During exponential growth, all of theseLactobacillus species acidified their growth medium at rates on the order of 106 protons/bacterium/s. Such rates, combined with an estimate of the total number of lactobacilli in the vagina, suggest that vaginal lactobacilli could reacidify the vagina at the rate observed postcoitally following neutralization by the male ejaculate (W. H. Masters and V. E. Johnson, Human sexual response, p. 93, 1966). During bacterial vaginosis (BV), there is a loss of vaginal acidity, and the vaginal pH rises to >4.5. This correlates with a loss of lactobacilli and an overgrowth of diverse bacteria. Three BV-associated bacteria, Gardnerella vaginalis, Prevotella bivia, andPeptostreptococcus anaerobius, acidified their growth medium to an asymptotic pH (4.7 to 6.0) consistent with the characteristic elevated vaginal pH associated with BV. Together, these observations are consistent with vaginal flora, rather than epithelial cells, playing a primary role in creating the acidity of the vagina.

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