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

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.

The Performance of Different AgTiO2 Loading into Poly(3-Nitrothiophene) for Efficient Adsorption of Hazardous Brilliant Green and Crystal Violet Dyes

The in-situ polymerization technology was used to successfully produce nanostructured binary nanocomposites (NCs) made from a poly (3-nitrothiophen) matrix (P3NT) that were loaded effectively with nanoparticles (NPs) of silver titanium dioxide (AgTiO2), of varying percentages (10%, 20%, and 30%). A uniform coating of P3NT covers the AgTiO2 NPs. Various methods were performed to confirm the fabrication of the binary P3NT/AgTiO2 NCs adsorbents, such as FTIR, XRD, SEM, and EDX. Both dyes (brilliant green (B.G.) and crystal violet (C.V.)) were removed from liquid media by using the binary P3NT/AgTiO2 NCs. A range of batch adsorption studies was used to optimize various factors that impact the elimination of B.G. or C.V. dyes, including the pH, weight of the binary P3NT/AgTiO2 NC, proportion of AgTiO2 NP, time, and temperature. The pseudo-second-order kinetics ( R 2 = 0.999 ) was better adapted for the adsorption procedure’s empirical data whereby the maximum adsorption capacity of the C.V. dye was 43.10 mg/g and ( R 2 = 0.996 ) the maximum adsorption potential was 40.16 mg/g for B.G. dye, succeeded by the pseudo-second-order kinetics. Moreover, the adhesion of B.G. and C.V. pigments on the layers of NCs involves an endothermic reaction. In addition, the concocted adsorbent not only exhibited strong adsorption characteristics during four consecutive cycles but also possessed a higher potential for its reuse. According to the findings, the NCs might possibly be used as a robust and reusable adsorbent to remove B.G. and C.V. pigments from an aqueous medium.