The opportunistic fungus Candida albicans is a component of denture plaque and is associated with denture-related stomatitis. Inter-kingdom interactions between C. albicans and bacteria exist in such multi-species biofilms, which may affect the microbial composition of the plaque. This study was performed to investigate the bacterial composition of denture plaques, and the correlation between the relative abundance of these bacteria and C. albicans. Thirty denture plaque and 16 dental plaque samples were collected from 18 denture wearers (mean age, 80.3 years). After DNA extraction, a meta 16S rDNA amplicon library was constructed using PCR primers targeting the V3-V4 hypervariable region of bacteria. The amplicon was evaluated by high-throughput sequencing, followed by bacterial population analysis. The concentrations of both C. albicans DNA and total bacterial DNA were determined by real-time PCR. The correlation between the relative abundance of each bacterial genus and C. albicans was analyzed through Spearman's rank correlation. The genera Streptococcus, Lactobacillus, Rothia, and Corynebacterium were found to be more abundant in dentures than in dental plaques. The predominant bacteria in healthcare-associated pneumonia also inhabited denture surfaces. Candida albicans was positively correlated with three acidogenic bacteria and negatively correlated with Leptotrichia and pathogens associated with periodontitis and endocarditis. Dentures may be significant reservoirs of pathogens causing aspiration pneumonia. Bacteria showing negative correlation with C. albicans, such as Leptotrichia, may be useful for controlling the growth of C. albicans in antifungal therapies.
Porphyromonas gingivalis , a Gram-negative oral anaerobe, is strongly associated with chronic adult periodontitis, and it utilizes FimA fimbriae to persistently colonize and evade host defenses in the periodontal crevice. The FimA-related gene cluster (the fim gene cluster) is positively regulated by the FimS-FimR two-component system. In this study, comparative analyses between fimbriate type strain ATCC 33277 and fimbria-deficient strain W83 revealed differences in their fimS loci, which encode FimS histidine kinase. Using a reciprocal gene exchange system, we established that FimS from W83 is malfunctional. Complementation analysis with chimeric fimS constructs revealed that W83 FimS has a defective kinase domain due to a truncated conserved G3 box motif that provides an ATP-binding pocket. The introduction of the functional fimS from 33277 restored the production, but not polymerization, of endogenous FimA subunits in W83. Further analyses with a fimA -exchanged W83 isogenic strain showed that even the fimbria-deficient W83 retains the ability to polymerize FimA from 33277, indicating the assembly of mature FimA by a primary structure-dependent mechanism. It also was shown that the substantial expression of 33277-type FimA fimbriae in the W83 derivative requires the introduction and expression of the functional 33277 fimS . These findings indicate that FimSR is the unique and universal regulatory system that activates the fim gene cluster in a fimA genotype-independent manner.
Antimicrobial photodynamic therapy (aPDT) is considered a potential treatment for biofilm infections, which have become an increasing health issue because of the rise in antimicrobial resistance. This study aimed to evaluate the bactericidal effect of aPDT using indocyanine green-loaded nanospheres with chitosan coating (ICG-Nano/c) against polymicrobial periodontal biofilms. Composite biofilms of Porphyromonas gingivalis and Streptococcus gordonii were constructed in 96-well plates, and aPDT with ICG-Nano/c and an 810 nm diode laser was performed either by direct irradiation or transmitting irradiation through a 3-mm-thick gingival model. The efficacy of ICG-Nano/c-based aPDT was compared with antibiotics (minocycline and amoxicillin). Additionally, attenuated aPDT under sublethal conditions was used to investigate gene expression related to the antioxidant response (oxyR and sod of P. gingivalis) and biofilm formation via quorum sensing (luxS of both species) with real-time polymerase chain reaction. ICG-Nano/c-based aPDT significantly reduced the bacterial load in the biofilm, achieving at least a 2 log10 reduction in colony-forming units within 5 minutes for both irradiation methods. After 6 hours of treatment, the bactericidal effects of aPDT and antibiotics were similar, but after 32 hours, antibiotics were more effective, particularly against P. gingivalis. Attenuated aPDT showed a slight increase in sod expression in P. gingivalis, while luxS expression decreased in both bacteria. The ICG-Nano/c-based aPDT system exerted a certain degree of bactericidal activity against a composite biofilm of periodontal bacteria. Therefore, it has potential as an alternative option or adjunctive therapy to conventional antibiotics in periodontal treatment.