Dual-species Biofilms Research Articles

Efficacy of chitosan root canal medicament against cross-kingdom dual-species biofilm of Candida albicans and Enterococcus faecalis in an in vitro root-canal model

Efficacy of chitosan root canal medicament against cross-kingdom dual-species biofilm of Candida albicans and Enterococcus faecalis in an in vitro root-canal model

Influence of Flagella on Salmonella Enteritidis Sedimentation, Biofilm Formation, Disinfectant Resistance, and Interspecies Interactions.

Flagella are essential for bacterial motility and biofilm formation by aiding bacterial attachment to surfaces. However, the impact of flagella on bacterial behavior, particularly biofilm formation, remains unclear. This study constructed two flagellar mutation strains of Salmonella Enteritidis (SE), namely, SE-ΔflhD and SE-ΔflgE, and confirmed the loss of flagellar structures and motility in these strains. The mutant strains exhibited growth comparable with the wild-type (WT) strain but had higher sedimentation rates. Biofilm biomass did not differ significantly between the WT and mutant strains, except for SE-ΔflgE at 3 d. SE-ΔflgE showed increased susceptibility to sodium hypochlorite compared to the WT. The co-sedimentation rate of flagella-deficient strains was lower than the WT, and the biomass of dual-species biofilm formed by Bacillus paramycoides B5 with SE-ΔflhD or SE-ΔflgE was significantly lower than with the WT. These findings emphasize the significance of SE flagella in biofilm formation and interspecies interactions, offering insights into targeted biofilm prevention and control measures.

Effect of Nd:YAG Laser Irradiation of the Growth of Oral Biofilm

Background: Oral microbiota comprises a wide variety of microorganisms. The purpose of this study was to evaluate the effects of Nd:YAG laser with a 1064 nm wavelength on the in vitro growth of Candida albicans, Candida glabrata, and Streptococcus mutans clinical strains, as well as their biofilm. The study also aimed to determine whether the parameters recommended for photobiomodulation (PBM) therapy, typically used for tissue wound healing, have any additional antibacterial or antifungal effects. Material and Methods: Single- and dual-species planktonic cell solution and biofilm cultures of Streptococcus mutans, Candida albicans, and Candida glabrata were irradiated using an Nd:YAG laser (LightWalker; Fotona; Slovenia) with a flat-top Genova handpiece. Two test groups were evaluated: Group 1 (G-T1) exposed to low power associated parameters (irradiance 0.5 W/cm2) and Group 2 (G-T2) with higher laser parameters (irradiance 1.75 W/cm2). Group 3 (control) was not exposed to any irradiation. The lasers’ effect was assessed both immediately after irradiation (DLI; Direct Laser Irradiation) and 24 h post-irradiation (24hLI) of the planktonic suspension using a quantitative method (colony-forming units per 1 mL of suspension; CFU/mL), and the results were compared with the control group, in which no laser was applied. The impact of laser irradiation on biofilm biomass was assessed immediately after laser irradiation using the crystal violet method. Results: Nd:YAG laser irradiation with photobiomodulation setting demonstrated an antimicrobial effect with the greatest immediate reduction observed in S. mutans, achieving up to 85.4% reduction at the T2 settings. However, the laser’s effectiveness diminished after 24 h. In single biofilm cultures, the highest reductions were noted for C. albicans and S. mutans at the T2 settings, with C. albicans achieving a 92.6 ± 3.3% reduction and S. mutans reaching a 94.3 ± 5.0% reduction. Overall, the T2 settings resulted in greater microbial reductions compared to T1, particularly in biofilm cultures, although the effectiveness varied depending on the microorganism and culture type. Laser irradiation, assessed immediately after using the crystal violet method, showed the strongest biofilm reduction for Streptococcus mutans in the T2 settings for both single-species and dual-species biofilms, with higher reductions observed in all the microbial samples at the T2 laser parameters (p < 0.05) Conclusion: The Nd:YAG laser using standard parameters typically applied for wound healing and analgesic effects significantly reduced the number of Candida albicans; Candida glabrata; and Streptococcus mutans strains.

Targeting mono- and dual-species biofilms of Staphylococcus aureus and Pseudomonas aeruginosa by the recombinant anticoagulant enzyme PAPC from micromycete Aspergillus ochraceus

Targeting mono- and dual-species biofilms of Staphylococcus aureus and Pseudomonas aeruginosa by the recombinant anticoagulant enzyme PAPC from micromycete Aspergillus ochraceus

Track of methylparaben in the bulk phase and on the extracellular matrix of dual-species biofilms: Biodegradation and bioaccumulation

Methylparaben (MP) is a preservative considered an environmental contaminant of emerging concern due to its persistence in water sources, including drinking water (DW). This study assesses the interaction between MP and dual-species biofilms of Acinetobacter calcoaceticus and Stenotrophomonas maltophilia. These biofilms were grown under realism-based conditions in a multiple-cylinder biofilm reactor on polypropylene (PPL) surfaces, for 7 days, and then exposed to MP at 0.5 mg/L for three consecutive days. S. maltophilia predominantly succeeds within these biofilms compared to A. calcoaceticus. Exposure to MP resulted in a 4-fold increase in the number of culturable cells and a 1.4-fold rise in polysaccharide content, suggesting that bacterial cells may utilize MP as a carbon source to enhance biofilm fitness. MP was found to adsorb to PPL with biofilms following a pseudo-second-order kinetic model. Circa 37 % of MP adsorbed to PPL after 3 days of exposure. Besides that, MP was biodegraded by biofilms following an apparent first-order kinetic model. Part (25 %) of the MP was biodegraded whereas only 0.02 % bioaccumulated on the biofilm matrix. Biodegradation was related to esterase and lipase activity. The results provide new insights into the interaction between MP with biofilms and materials used in DW industries.

Effect of Pipe Materials and Interspecific Interactions on Biofilm Formation and Chlorine Resistance: Turn Enemies into Friends

Drinking water distribution systems (DWDSs) may be contaminated to various degrees when different microorganisms attach to the pipe walls. Understanding the characteristics of biofilms on pipe walls can help prevent and control microbial contamination in DWDSs. The biofilm formation, interspecific interactions, and chlorine resistance of 10 dual-species biofilms in polyethylene (PE) and cast iron (CI) pipes were investigated in this paper. The biofilm biomass (heterotrophic bacterial plate count and crystal violet) of dual species in CI pipes is significantly higher than that in PE pipes, but the biofilm activity in CI pipes is significantly lower than that in PE pipes. The interspecific interaction of Sphingomonas-containing group presented synergistic or neutral relationship in PE pipes, whereas the interspecific interaction of the Acidovorax-containing group showed a competitive relationship in CI pipes. Although interspecific relationships may help bacteria resist chlorine, the chlorine resistance was more reliant on dual-species groups and pipe materials. In CI pipes, the Microbacterium containing biofilm groups showed better chlorine resistance, whereas in PE pipes, most biofilm groups with Bacillus exhibited better chlorine resistance. The biofilm groups with more extracellular polymeric substance (EPS) secretion showed stronger chlorine resistance. The biofilm in the PE pipe is mainly protected by EPS, while both EPS and corrosion products shield the biofilms within CI pipe. These results supported that dual-species biofilms are affected by pipe materials and interspecific interactions and provided some ideas for microbial control in two typical pipe materials.

SDF, KI, and NSF antibacterial effects on dual-species biofilm

SDF, KI, and NSF antibacterial effects on dual-species biofilm

Evaluation of riboflavin, nanocurcumin, and hydrogen peroxide under light conditions: reduction of mature dental biofilms and enamel mineral loss

BackgroundBiofilms are a potential harbor for many microorganisms. The aim of this study was to test the efficacy of riboflavin (Rib), nano-micelle curcumin (NC), and hydrogen peroxide (HP), alone and in combination with the respective light (light-emitting diode (LED) or 980 nm diode laser) on the reduction of Streptococcus mutans and Lactobacillus acidophilus dual-species biofilms and their effect on the enamel mineral loss. Materials and methodsThe biofilms were formed on saliva-coated enamel slabs. Then, the biofilms were treated with antimicrobial photodynamic therapy (PDT) based on LED, Rib, and NC photosensitizers and with HP also based on a 980 nm diode laser (n = 8 per group). A crystal violet assay was performed to determine the reduction of the dual-species biofilms. The enamel slabs were analyzed for calcium and phosphorus content by scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX). ResultsWhile HP-PDT showed a reduction of 37% (p < 0.001), PDT with NC resulted in an even greater reduction of dual-species biofilms (40%, p < 0.001) than HP- and Rib-mediated PDT. In the EDX test, no significant difference was found between the control group and the treatment groups. ConclusionsThe use of natural photosensitizers such as NC in PDT has an effect that may be potentially important in reducing caries-causing bacteria.

Veillonella parvula acts as a pathobiont promoting the biofilm virulence and cariogenicity of Streptococcus mutans in adult severe caries.

Adult severe caries (ASC) brings severe oral dysfunction and treatment difficulties to patients, and yet no clear pathogenic mechanism for it has been found. This study is focused on the composition of dental plaque microbiome profiles in order to identify disease-relevant species and to investigate into their interactions with the S. mutans. Samples of dental plaque were collected for metagenomic analysis. The acidification, aciduricity, oxidative stress tolerance, and gtf (glucosyltransferase) gene expression of S. mutans cocultured with V. parvula which was identified as ASC-related dominant bacterium. The biofilm formation and extracellular exopolysaccharide (EPS) synthesis of dual-strain were analyzed with scanning electron microscopy (SEM), crystal violet (CV) staining, live/dead bacterial staining, and confocal laser scanning microscopy (CLSM). Furthermore, rodent model experiments were performed to validate the in vivo cariogenicity of the dual-species biofilm. The most significantly abundant taxon found associated with ASC was V. parvula. In vitro experiments found that V. parvula can effectively promote S. mutans mature biofilm formation with enhanced acid resistance, hydrogen peroxide detoxicity, and biofilm virulence. Rodent model experiments revealed that V. parvula was incapable of causing disease on its own, but it significantly heightened the biofilm virulence of S. mutans when being co-infected and augmented the progression, quantity, and severity of dental caries. Our findings demonstrated that V. parvula may act as a synergistic pathobiont to modulate the metabolic activity, spatial structure, and pathogenicity of biofilms of S. mutans in the context of ASC.IMPORTANCEAdult severe caries (ASC), as a special type of acute caries, is rarely reported and its worthiness of further study is still in dispute. Yet studies on the etiology of severe caries in adults have not found a clear pathogenic mechanism for it. Knowledge of the oral microbiota is important for the treatment of dental caries. We discovered that the interaction between V. parvula and S. mutans augments the severity of dental caries in vivo, suggesting V. parvula may act as a synergistic pathobiont exacerbating biofilm virulence of S. mutans in ASC. Our findings may improve the understanding of ASC pathogenesis and are likely to provide a basis for planning appropriate therapeutic strategies.

Klebsiella pneumoniae AI-2 transporters mediate interspecies interactions and composition in a three-species biofilm community

Biofilms in nature often exist as communities. In this study, an experimental mixed-species community consisting of Pseudomonas aeruginosa, Pseudomonas protegens and Klebsiella pneumoniae was used to investigate how AI-2 transporters affect interspecies interactions and composition. The K. pneumoniae lsrB/lsrD deletion mutants had a 10–25-fold higher concentration of extracellular AI-2 compared to the wild-type. Although these deletion mutants produced monospecies biofilms of similar biomass, the substitution of these mutants for the parental strain significantly altered composition. Dual-species biofilm assays demonstrated that the changes in composition were due to the cumulative effect of pairwise interactions. It was further revealed that K. pneumoniae being present physically in the consortium was important in AI-2 mediating composition in the consortium, and that AI-2 transporters were crucial in achieving maximum biomass in the community. In conclusion, these findings demonstrate that AI-2 transporters mediate interspecies interactions and is important in maintaining the compositional equilibrium of the community.

Assessment of the Antibiofilm Performance of Silver-Containing Wound Dressings: A Dual-Species Biofilm Model.

Background It is commonly accepted that microorganisms found within hard-to-heal wounds are present in biofilm form. Biofilms are often polymicrobial in nature, which increases their virulence and tolerance to antimicrobial agents. The aim of this study was to compare the antibiofilm activity of silver-containing antimicrobial wound dressings in a dual-species simulated wound biofilm model. Materials and methods Four silver-containing wound dressings were evaluated in vitro: Aquacel® Ag+ Extra™ dressing, KerraContact® Ag dressing, Durafiber* Ag dressing, and UrgoClean Ag dressing. Each dressing was applied to a simulated wound assembly containing biofilm-gauze inoculated with Klebsiella pneumoniae and methicillin-resistant Staphylococcus aureus (MRSA). Each biofilm-inoculated gauze was incubated at 35±3ºC for 6, 24, 48 and 72 hours. Enumeration of surviving biofilm bacteria at each time point was performed in triplicate for each test dressing and its equivalent control. Results Aquacel® Ag+ Extra™ dressing was observed to reduce the biofilm population within 24 hours with a >4 log10 kill observed for K. pneumoniae and >6 log10 for MRSA from an initial biofilm challenge of 4.16×109 CFU/mL. This kill rate was sustained for the duration of the challenge period, with Aquacel®Ag+ Extra™ dressing reducing the biofilm population to non-detectable levels (<30 Colony Forming Units (CFU) per test) by 72 hours for K. pneumoniae and by 48 hours for MRSA. KerraContact® Ag dressing demonstrated an initial reduction at 6 hours of ~2 log10 in both K. pneumoniae and MRSA. Durafiber* Ag dressing exhibited a slight, gradual reduction in biofilm population over the course of the test period, reducing each challenge organism by ~2.5 log10 by 72 hours. UrgoClean Ag was shown to have little to no impact on the dual-species biofilm with levels remaining similar or greater than that recovered prior to dressing application. The no-dressing biofilm-colonised gauze control demonstrated that the biofilm bacteria remained viable throughout the test period and species population proportionality was maintained. Conclusion Using a dual-species simulated wound biofilm model comprising the pathogens K. pneumoniae and MRSA, Aquacel® Ag+ Extra™ dressing demonstrated significantly greater antibiofilm activity than the other silver-containing dressings. The enhanced antibiofilm activity of Aquacel® Ag+ Extra™ dressing in this study may be attributed to the additional antibiofilm agents, ethylenediaminetetraacetic acid and benzethonium chloride, contained within the dressing.

Coaggregation dynamics in drinking water biofilms and implications for chlorine disinfection

Biofilms in drinking water (DW) systems persistently challenge traditional disinfection methods due to intricate microbial interactions, with coaggregation playing a crucial role in forming multispecies biofilms. This study examined the implications of coaggregation on tolerance towards sodium hypochlorite (NaOCl) disinfection. Dual-species biofilms were formed for seven days on polyvinyl chloride coupons, comprising a strain of the emerging pathogen Stenotrophomonas maltophilia and the coaggregating strain Delftia acidovorans 005 P. For comparison, dual-species biofilms were also formed with a non-coaggregation strain (D. acidovorans 009 P). The minimum bactericidal concentration (MBC) for each planktonic strain varied (D. acidovorans: 1 mg/L, S. maltophilia: 1.5 mg/L) below the safe DW treatment limits. However, high NaOCl doses (10 ×MBC and 100 ×MBC,) showed low efficacy against dual-species biofilms, indicating significant biofilm tolerance to disinfection. Membrane damage occurred at sub-MBC without culturability loss, underscoring biofilm resilience. The biofilm analysis revealed a complex interplay between the composition of extracellular polymeric substances and the architecture, which was influenced by the presence of the coaggregating strain. Overall, coaggregation significantly influenced biofilm formation and resilience, impacting NaOCl disinfection. These findings underscore the challenges of microbial interactions in biofilms, emphasizing the need for improved disinfection strategies to control biofilms in drinking water systems.

Synergistic effects of quorum-sensing molecules and antimicrobials against Candida albicans and Pseudomonas aeruginosa biofilms: in vitro and in vivo studies.

Candida albicans can form polymicrobial biofilms with other microorganisms, such as Pseudomonas aeruginosa, at infection sites. As biofilms are highly resistant to antibiotics there is a need for new antibiofilm agents that have unique targets and modes of action. In this study the antibiofilm effects of two quorum-sensing molecules (QSMs), farnesol and tyrosol, were investigated alone and in combination with antibiotics (aztreonam, colistin, tobramycin) and antifungals (fluconazole, amphotericin B, caspofungin), against single- and dual-species biofilms of C. albicans and P. aeruginosa in in vitro and in vivo systems. It was observed that QSMs alone, especially farnesol, showed at least a 1-log reduction against preformed single- and dual-species biofilms of C. albicans and P. aeruginosa. Combination of QSMs with colistin or fluconazole was found to be effective against both single- and dual-species biofilms in vitro. Increased survival was observed in C. elegans when treated with colistin or fluconazole in combination with QSMs, compared with no treatment. Additionally, the QSMs and colistin and farnesol combinations effectively inhibited biofilm formation by C. albicans and P. aeruginosa on bronchial epithelial cells, and reduced IL-1β expression in lung bronchial epithelial cells. There is a need for effective treatments for bacterial-fungal biofilm infections and, to our knowledge, there have been no studies of QSMs and antimicrobial combinations against dual-species biofilms involving C. albicans and P. aeruginosa. Hence these findings will make a significant contribution to the literature.

Inter-Species Competition of Mono- or Dual Species Biofilms- of MDR-Staphylococcus aureus and Pseudomonas aeruginosa Promotes the Killing Efficacy of Phage or Phage Cocktail

Pseudomonas aeruginosa and Staphylococcus aureus are opportunistic bacteria frequently linked to burn wound infections. These bacteria can grow as biofilms, which increases their level of drug resistance to current antibiotics. The purpose of the present study is to analyze the effect of biofilm formation, phage and phage cocktail action on single species and dual species biofilms I, e the coexistence of Gram positive (S. aureus) and Gram negative (P. aeruginosa). To this scenario, we employed multi-drug resistant bacteria (P. aeruginosa and S. aureus at 109 CFU/µL) biofilm as single and in combination of both Gram-positive and Gram-negative bacterial biofilms of 24 h grown with respective phage (109 PFU/µL) and phage cocktail (109 PFU/µL) at 4 h of incubation under static conditions. The bacteriolytic activity of phages vB_SAnS_SADP1 and vB_PAnP_PADP4 on 24-h-old biofilms of P. aeruginosa (0.761 ± 0.031) and S. aureus (0.856 ± 0.055), both alone and in combination (0.67 ± 0.02), was the focus of this investigation. The structural organization of biofilms in single- or dual-species combinations under in vitro conditions was validated by scanning and confocal laser scanning microscopy investigations. After 24 h of incubation, single-species biofilms are denser and more resilient whereas dual species biofilms are more loosely associated. Loose association of dual-species biofilm under scanning electron microscopic images at the same conditions, indicated the interspecies -competition of the Gram-positive and Gram-negative bacteria and dual-species biofilms (0.67 ± 0.02) have weak associations and are readily impacted by phage and a phage cocktail (0.16 ± 0.02). Dual-species biofilms were more readily impacted in in vitro settings.

NIR-II triggered Cu(I) phosphide for chemodynamic and photothermal periodontitis treatment: Efficient reduction of bacterial co-aggregation

The synergy between chemodynamic therapy (CDT) and photothermal therapy (PTT) offers a promising antimicrobial strategy for periodontitis, yet faces challenges like complex material structure and limited NIR-I light penetration. Additionally, low endogenous H2O2 levels in biofilm and a focus on bacterial eradication over colonization prevention limit current treatments. To address these issues, we newly introduce a single-material system (Cu3P@PAH@Lox) that integrates dual functionalities to synergistically enhance antimicrobial effects and significantly reduce pathogen co-aggregation. This system utilizes PTT to increase local temperature, boosting •OH production in CDT while downregulating heat shock proteins to enhance PTT efficacy, forming a self-reinforcing feedback loop. Lactate oxidase (Lox) is employed to convert lactate—a metabolite in periodontal biofilm—into H2O2, further amplifying CDT's potential. In vitro Cu3P@PAH@Lox demonstrates a remarkable synergistic effect against dual-species biofilms by more than 2-log reduction of colony-forming unit. Moreover, Cu3P@PAH@Lox exhibits outstanding synergistic antibacterial performances to alleviate inflammation and destruction of tissue in vivo periodontitis model. Furthermore, the mechanism of pathogen co-aggregation disruption by PTT is verified via the Cbe-Ltp1-Ptk1-fimA signaling pathway. This single-material multimodal system we have herein demonstrated for the first time marks a significant advancement in periodontitis treatment, eradicating microbes and preventing bacterial colonization, offering a path to comprehensive periodontal care. Statement of significanceThe synergy between chemodynamic therapy (CDT) and photothermal therapy (PTT) has been considered a promising therapy for periodontitis. Yet, facing challenges, the complex material structure, limited NIR-I light penetration, low endogenous H2O2 level in biofilm, and a focus on bacterial eradication over colonization prevention are still insufficient. This study pioneers a unique, single-material system (Cu3P@PAH@Lox) that synergistically enhances antimicrobial effects and substantially curtails pathogen co-aggregation, advancing periodontitis therapy. By exploiting PTT to elevate local temperatures, thereby increasing hydroxyl radical production in CDT and concurrently suppressing heat shock proteins, the system establishes a potent, self-enhancing loop. Furthermore, lactate oxidase is innovatively utilized to convert lactate from periodontal biofilm into hydrogen peroxide, augmenting the efficacy of CDT. The introduction of Cu3P@PAH@Lox is poised to revolutionize periodontitis treatment, eliminating microbes and impeding bacterial colonization, thereby charting a course for comprehensive periodontal management.

Cocktail Approach with Polyserotonin Nanoparticles and Peptides for Treatment of Streptococcus mutans.

Dental plaque, formed by a Streptococcus mutans biofilm, is a major contributor to cavity formation. While antimicrobial strategies exist, the growing risk of antibiotic resistance necessitates alternative therapeutic solutions. Polyserotonin nanoparticles (PSeNPs), recently recognized for their photothermal property and promising biomedical applications, open up a new avenue for antimicrobial use. Here, we introduced a UV-initiated synthetic route for PSeNPs with improved yield. Using these PSeNPs, a cocktail treatment to reduce the viability of this cavity-causing bacteria was developed. This cocktail comprises an S. mutans-targeting antimicrobial peptide (GH12), an intraspecies competence-stimulating peptide that triggers altruistic cell death in S. mutans, and laser-activated heating of PSeNPs. The "peptide + PSeNP + laser" combination effectively inhibits S. mutans growth in both planktonic and biofilm states. Moreover, the cocktail approach remains effective in reducing the viability of S. mutans in a more virulent dual-species biofilm with Candida albicans. Overall, our results reinforce the utility of a multipronged therapeutic strategy to reduce cariogenic bacteria in the complex model oral biofilm.

Inhibition of Development and Metabolism of Dual-Species Biofilms of Candida albicans and Candida krusei (Pichia kudriavzevii) by Organoselenium Compounds.

Although Candida albicans is the most frequently identified Candida species in clinical settings, a significant number of infections related to the non-albicans Candida (NAC) species, Candida krusei, has been reported. Both species are able to produce biofilms and have been an important resistance-related factor to antimicrobial resistance. In addition, the microbial relationship is common in the human body, contributing to the formation of polymicrobial biofilms. Considering the great number of reports showing the increase in cases of resistance to the available antifungal drugs, the development of new and effective antifungal agents is critical. The inhibitory effect of Organoselenium Compounds (OCs) on the development of Candida albicans and Candida krusei was recently demonstrated, supporting the potential of these compounds as efficient antifungal drugs. In addition, OCs were able to reduce the viability and the development of biofilms, a very important step in colonization and infection caused by fungi. Thus, the objective of this study was to investigate the effect of the Organoselenium Compounds (p-MeOPhSe)2, (PhSe)2, and (p-Cl-PhSe)2 on the development of dual-species biofilms of Candida albicans and Candida krusei produced using either RPMI-1640 or Sabouraud Dextrose Broth (SDB) media. The development of dual-species biofilms was evaluated by the determination of both metabolic activity, using a metabolic assay based on the reduction of XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide sodium salt) assay and identification of either Candida albicans and Candida krusei on CHROMagar Candida medium. Biofilm formation using RPMI-1640 was inhibited in 90, 55, and 20% by 30 µM (p-MeOPhSe)2, (PhSe)2, and (p-Cl-PhSe)2, respectively. However, biofilms produced using SDB presented an inhibition of 62, 30 and 15% in the presence of 30 µM (p-MeOPhSe)2, (PhSe)2, and (p-Cl-PhSe)2, respectively. The metabolic activity of 24 h biofilms was inhibited by 35, 30 and 20% by 30 µM (p-MeOPhSe)2, (PhSe)2, and (p-Cl-PhSe)2, respectively, with RPMI-1640; however, 24 h biofilms formed using SDB were not modified by the OCs. In addition, a great reduction in the number of CFUs of Candida albicans (93%) in biofilms produced using RPMI-1640 in the presence of 30 µM (p-MeOPhSe)2 was observed. However, biofilms formed using SDB and treated with 30 µM (p-MeOPhSe)2 presented a reduction of 97 and 69% in the number of CFUs of Candida albicans and Candida krusei, respectively. These results demonstrated that Organoselenium Compounds, mainly (p-MeOPhSe)2, are able to decrease the metabolic activity of dual-species biofilms by reducing both Candida albicans and Candida krusei cell number during biofilm formation using either RPMI-1640 or SDB. Taken together, these results demonstrated the potential of the OCs to inhibit the development of dual-species biofilms of Candida albicans and Candida krusei.

The arrangement of dual-species biofilms of Candida albicans and Issatchenkia orientalis can be modified by the medium: effect of Voriconazole

Both Candida albicans and Issatchenkia orientalis have been isolated from different types of infections over the years. They have the ability to form communities of microorganisms known as biofilms. It has been demonstrated that the medium employed in studies may affect the biofilm development. The aim of this study was to investigate the arrangement of dual-species biofilms of C. albicans and I. orientalis cultivated on either RPMI-1640 or Sabouraud Dextrose Broth (SDB), as well as the inhibitory effect of Voriconazole (VRC). For the experiments performed, ATCC strains were used, and yeast-mixed suspensions were inoculated in 96-well plates with either RPMI-1640 or SDB, in the presence or absence of VRC. The results were observed by counting the number of CFU obtained from scraping off the biofilms produced and plating the content on CHROMagar Candida medium. It was observed that for all conditions tested the medium chosen affected the arrangement of dual-species biofilms: when RPMI-1640 was used, there was a prevalence of C. albicans, while the opposite was noted when SDB was used. It could be suggested that the medium and environment could regulate interactions between both yeast species, including the response to different antifungal drugs.

Antimicrobial Tolerance in Cross-Kingdom Dual-Species Biofilms Formed by Fungi and Bacteria.

Candida albicans, the most common pathogenic fungus, can form biofilms on the surface of medical devices and often causes bloodstream infections. Biofilms have a complex structure composed of microorganisms and a surrounding extracellular matrix. Biofilms are difficult to treat because they are resistant to antifungal drugs and the host environment. Nearly one in four patients with candidemia have a polymicrobial infection. These polymicrobial biofilms, especially those comprising cross-kingdom species of fungi and bacteria, can lead to long hospital stays and high mortality rates. This review outlines the unique interactions of dual-species biofilms with Candida albicans and the clinically important bacteria Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli.

Destruction of cariogenic biofilms by antimicrobial photodynamic therapy mediated by phycocyanin and toluidine blue along with sodium fluoride varnish or titanium tetrafluoride

ObjectiveEvaluation of the effect of phycocyanin (PC) and toluidine blue (TBO) along with sodium fluoride varnish (FV) or titanium tetrafluoride (TiF4) under the conditions of antimicrobial photodynamic therapy (PDT) on a dual-species cariogenic biofilm and on remineralization process. DesignAfter the development of Streptococcus mutans and Lactobacillus acidophilus dual-species biofilms on the human enamel disks, they were divided into 11 groups (n = 9): Control (0.9 % saline), PC, TBO, FV, and TiF4 alone, PC and TBO in combination with a 635 nm diode laser (PDT treatment), PC-PDT+ (PC + FV or TiF4 + 635 nm diode laser), and TBO-PDT+ (TBO + FV or TiF4 + 635 nm diode laser). After the treatment, crystal violet assay was performed to determine the reduction of cariogenic biofilms. Enamel remineralization changes were analyzed using energy dispersive X-ray spectroscopy (EDX) and field emission scanning electron microscopy (FESEM) for the calcium and phosphorus (Ca/P) ratio. ResultsOnly TBO-PDT+ showed superior antibiofilm activity when TiF4 was applied. Furthermore, the highest Ca/P ratio was found after treatment of enamel surfaces with TiF4-TBO-PDT+. The FESEM images showed that the enamel disks treated with TiF4 plus TBO-mediated PDT exhibited surface coating. However, TiF4 plus PC-mediated PDT cannot repair demineralized enamel. ConclusionsThese data suggest that TBO-PDT along with TiF4 can effectively reduce cariogenic biofilms and significantly remineralize enamel disks, opening new avenues in caries prevention.