Biofilm Virulence Research Articles

Antibacterial potential of some seaweeds species to combat biofilm-producing multi-drug resistant Staphylococcus aureus of Nile tilapia

THERE is an increasing worldwide demand for seafood due to the awareness of fish as an important protein source for a growing population. However, the diseases caused by multi-drug resistant (MDR) bacterial infections and the low survival rate of the fish represent significant challenges to fish farmers. The virulence and pathogenicity of bacteria are often enhanced when growing as a biofilm. Therefore, a variety of new antimicrobial drugs has attracted wide attention in treating fish pathogen bacteria. Herein, the efficiency of macroalgal extracts as an antibacterial agent against MDR Staphylococcus aureus bacteria isolated from Nile tilapia (Oreochromis niloticus) was studied. Out of 200 bacterial isolates, 50 strains were identified as S. aureus. Of these strains, 37 were MDR and showed a potential role in the production of virulence factors, including staphylokinase (70.2%), lecithinase (81%), protease (56.7%) and lipase (59.4%). Significant production of biofilm virulence factor by MDR S. aureus strains was also observed from the quantitative and qualitative analysis. Four algal species namely Jania rubens, Ulva lactuca, Sargassum vulgare, and Sargassum fusiforme were tested for their antibacterial activity against MDR S. aureus strains. Of those, S. vulgare diethyl ether extract showed the highest antibacterial. In addition, GC-MS analysis revealed 20 identified components in S. vulgare diethyl ether extract, in which Longifolene was dominant (16.5%). This study thus established the possibility of developing an antibacterial agent to combat developing MDR S. aureus and biofilm-related infections in Nile tilapia.

Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans.

N-Acetylglucosamine (GlcNAc) and glucosamine (GlcN) enhance the competitiveness of the laboratory strain DL1 of Streptococcus gordonii against the caries pathogen Streptococcus mutans Here, we examine how amino sugars affect the interaction of five low-passage-number clinical isolates of abundant commensal streptococci with S. mutans by utilizing a dual-species biofilm model. Compared to that for glucose, growth on GlcN or GlcNAc significantly reduced the viability of S. mutans in cocultures with most commensals, shifting the proportions of species. Consistent with these results, production of H2O2 was increased in most commensals when growing on amino sugars, and inhibition of S. mutans by Streptococcus cristatus, Streptococcus oralis, or S. gordonii was enhanced by amino sugars on agar plates. All commensals except S. oralis had higher arginine deiminase activities when grown on GlcN and, in some cases, GlcNAc. In ex vivo biofilms formed using pooled cell-containing saliva (CCS), the proportions of S. mutans were drastically diminished when GlcNAc was the primary carbohydrate. Increased production of H2O2 could account in large part for the inhibitory effects of CCS biofilms. Surprisingly, amino sugars appeared to improve mutacin production by S. mutans on agar plates, suggesting that the commensals have mechanisms to actively subvert antagonism by S. mutans in cocultures. Collectively, these findings demonstrate that amino sugars can enhance the beneficial properties of low-passage-number commensal oral streptococci and highlight their potential for moderating the cariogenicity of oral biofilms.IMPORTANCE Dental caries is driven by dysbiosis of oral biofilms in which dominance by acid-producing and acid-tolerant bacteria results in loss of tooth mineral. Our previous work demonstrated the beneficial effects of amino sugars GlcNAc and GlcN in promoting the antagonistic properties of a health-associated oral bacterium, Streptococcus gordonii, in competition with the major caries pathogen Streptococcus mutans Here, we investigated 5 low-passage-number clinical isolates of the most common streptococcal species to establish how amino sugars may influence the ecology and virulence of oral biofilms. Using multiple in vitro models, including a human saliva-derived microcosm biofilm, experiments showed significant enhancement by at least one amino sugar in the ability of most of these bacteria to suppress the caries pathogen. Therefore, our findings demonstrated the mechanism of action by which amino sugars may affect human oral biofilms to promote health.

Effect of D-cysteine on dual-species biofilms of Streptococcus mutans and Streptococcus sanguinis

Dental caries is a highly prevalent disease worldwide. It is caused by the cariogenic biofilms composed of multiple dynamic bacteria on dental surface. Streptococcus mutans and Streptococcus sanguinis are resident members within the biofilms and an antagonistic relationship has been shown between these two species. S. mutans, as the major causative microorganism of dental caries, has been reported to be inhibited by free D-cysteine (D-Cys). However, whether D-Cys could affect S. sanguinis and the interspecies relationship between S. mutans and S. sanguinis remains unknown. The aim of the current study was to investigate the effect of D-Cys on the growth and cariogenicity of dual-species biofilms formed by S. mutans and S. sanguinis. We measured dual-species biofilms biomass, metabolic activity, lactate production. We also detected the biofilms structure, the ratio of live/dead bacteria, extracellular polysaccharide (EPS) synthesis and bacterial composition in the dual-species biofilms. We found that D-Cys could reduce the metabolic activity and lactic acid production of dual-species biofilms (p < 0.05). In addition, biofilms formation, the proportion of S. mutans in dual-species biofilms, and EPS synthesis were decreased with D-Cys treatment. The results suggested that D-Cys could inhibit the growth and cariogenic virulence of dual-species biofilms formed by S. mutans and S. sanguinis, indicating the potential of D-Cys in clinical application for caries prevention and treatment.

Voriconazole inhibits cross-kingdom interactions between Candida albicans and Actinomyces viscosus through the ergosterol pathway

Candida albicans and Actinomyces viscosus are prominent microbes associated with dental root caries. The aim of this study was to investigate the effect of C. albicans on A. viscosus biofilms and to identify the mechanisms associated with this interaction. A. viscosus and C. albicans strains (wide-type and mutants) were used to form biofilms in vitro and in vivo, which were subsequently analysed by crystal violet assay and scanning electron microscopy (SEM) to investigate the effect of C. albicans on A. viscosus growth. A viable plate count and survival curve for C. albicans mutants and A. viscosus combinations were used to identify which C. albicans pathway was crucial for cross-kingdom interactions. Voriconazole was used to block their interactions both in vitro and in vivo. SEM, fluorescence in situ hybridisation (FISH), quantitative PCR and survival curve analyses were performed to evaluate the activity of voriconazole on C. albicans and A. viscosus interactions. The biomass and virulence of mixed-species biofilms were significantly enhanced compared with the A. viscosus biofilm alone. However, this was not observed in the mixed-species biofilms with the C. albicans mutant erg11Δ/Δ in vitro and in vivo, indicating that azoles may work on the mixed-species biofilms. As expected, voriconazole can effectively reduce the biomass of mixed-species biofilms. A high concentration of voriconazole (1 µg/mL) reduced the abundance of C. albicans, whilst a low voriconazole concentration (0.25 µg/mL) blocked their interactions similar to the effect of the erg11Δ/Δ mutant. Voriconazole may be a candidate strategy to combat root caries pathogens.

Cross-kingdom interaction of Candida albicans and Actinomyces viscosus elevated cariogenic virulence

ObjectiveHow the interactions between Candida albicans and Actinomyces viscosus contributed to the root caries was not clear. This study aimed to investigate their cross-kingdom interactions on the biomass and the cariogenic virulence in dual-species biofilms. DesignSuspensions of C. albicans and A. viscosus were formed the mono and polymicrobial biofilms in vitro. Crystal violet assay, viable plate count, scanning electron microscopy and fluorescence in situ hybridization were used to analyze the biomass and biofilm structure. Glycolytic pH drop and the spectrophotometric method were used to evaluate the acid production and hydroxyapatite dissolution, respectively. The exopolysaccharide production was measured by the anthrone-sulfuric acid method, while the adhesion force was measured by atomic force microscopy. ResultsThe biomass and colony-forming units of mixed-species were significantly increased compared to that of the mono-species at 24 h, 48 h, 72 h. The structure of dual-species biofilm had more microcolonies and was much denser. The dual-species biofilms significantly decreased the pH value and damaged the hydroxyapatite compared with the mono-species biofilms at various time points, indicating the strong cariogenic virulence. Moreover, the dual-species biofilms significantly enhanced the exopolysaccharide production and adhesion force suggesting the increase of biofilm adhesion. ConclusionsCross-kingdom interactions of C. albicans and A. viscosus significantly elevated the biomass and cariogenic virulence of dual-species biofilm.

Association of biofilm formation and cytotoxic potential with multidrug resistance in clinical isolates of Pseudomonas aeruginosa.

Multidrug resistant (MDR) Pseudomonas aeruginosa having strong biofilm potential and virulence factors are a serious threat for hospitalized patients having compromised immunity. In this study, 34 P. aeruginosa isolates of human origin (17 MDR and 17 non-MDR clinical isolates) were checked for biofilm formation potential in enriched and minimal media. The biofilms were detected using crystal violet method and a modified software package of the automated VideoScan screening method. Cytotoxic potential of the isolates was also investigated on HepG2, LoVo and T24 cell lines using automated VideoScan technology. Pulse field gel electrophoresis revealed 10 PFGE types in MDR and 8 in non-MDR isolates. Although all isolates showed biofilm formation potential, strong biofilm formation was found more in enriched media than in minimal media. Eight MDR isolates showed strong biofilm potential in both enriched and minimal media by both detection methods. Strong direct correlation between crystal violet and VideoScan methods was observed in identifying strong biofilm forming isolates. High cytotoxic effect was observed by 4 isolates in all cell lines used while 6 other isolates showed high cytotoxic effect on T24 cell line only. Strong association of multidrug resistance was found with biofilm formation as strong biofilms were observed significantly higher in MDR isolates (p-value < 0.05) than non-MDR isolates. No significant association of cytotoxic potential with multidrug resistance or biofilm formation was found (p-value > 0.05). The MDR isolates showing significant cytotoxic effects and strong biofilm formation impose a serious threat for hospitalized patients with weak immune system.

The Prospects of a Highly Virulent Biofilm Dispersing Bacteriophage Isolated From Environment for Controlling &lt;i&gt;Enterobacter cloacae&lt;/i&gt; in Clinical Medicine

The global emergence of multi-drug resistant Enterobacter cloacae complex is one of the critical threats faced today by health care system as indicated by World Health Organisation. E. cloacae is one of the most common Enterobacter species amongst the E. cloacae complex and is often found closely associated with Enterobacter asburiae. The recent emergence of carbapenem-resistant E. cloacae complex against last-resort carbapenems has caught the attention of medical professionals and researchers who are now looking for therapeutic alternatives. Multidrug-resistant (MDR) E. cloacae complex is responsible for a high mortality rate amongst hospitalized patients frequently causing bacteraemia, urinary tract infections, lower respiratory tract infections and others. Exploring alternate antibacterial therapeutic strategies like bacteriophages may be of paramount importance. The present study describes a highly virulent bacteriophage N5822, isolated from an environmental source displaying high lytic activity towards E. cloacae / E. asburiae. N5822 bacteriophage has a short burst time and high burst size when compared to other known Enterobacter phages. N5822 bacteriophage holds its lytic activity over several generations of exposures to the host. N5822 bacteriophage was found to be efficient in reduction of preformed static host biofilm, as well as inhibits the formation of new biofilm by up to 90%. It displayed high lytic activity, specificity, stable virulence and high biofilm dispersive capacity against the host showing great promise as antibacterial for E.cloacae / E. asburiae. While some studies today are focusing on genetically engineered phages to tackle the menace of MDR pathogens including biofilm producing MDR pathogens, we worry about the escape of such genetically engineered organisms into the environment may influence the bacterial community dynamics, bacterial genome evolution and ecosystem biogeochemistry. By employing improved and efficient phage isolation methods, the natural abundance of bacteriophages in the environment can be tapped for the greater purpose of human health. Funding Statement: This work was funded by the Department of Science and Technology – Science and Engineering Research Board (DST-SERB) [grant number GAP-1-2066] and National Mission for Clean Ganga – Namami Gange Project (G – 2103). Also, the Council of Scientific and Industrial Research (CSIR) for funded the research scholar. Declaration of Interests: The authors declare they have no conflict of interest.

Novel Approaches to the Control of Oral Microbial Biofilms.

Effective management of biofilm-related oral infectious diseases is a global challenge. Oral biofilm presents increased resistance to antimicrobial agents and elevated virulence compared with planktonic bacteria. Antimicrobial agents, such as chlorhexidine, have proven effective in the disruption/inhibition of oral biofilm. However, the challenge of precisely and continuously eliminating the specific pathogens without disturbing the microbial ecology still exists, which is a major factor in determining the virulence of a multispecies microbial consortium and the consequent development of oral infectious diseases. Therefore, several novel approaches are being developed to inhibit biofilm virulence without necessarily inducing microbial dysbiosis of the oral cavity. Nanoparticles, such as pH-responsive enzyme-mimic nanoparticles, have been developed to specifically target the acidic niches within the oral biofilm where tooth demineralization readily occurs, in effect controlling dental caries. Quaternary ammonium salts (QAS) such as dimethylaminododecyl methacrylate (DMADDM), when incorporated into dental adhesives or resin composite, have also shown excellent and durable antimicrobial activity and thus could effectively inhibit the occurrence of secondary caries. In addition, custom-designed small molecules, natural products and their derivatives, as well as basic amino acids such as arginine, have demonstrated ecological effects by modulating the virulence of the oral biofilm without universally killing the commensal bacteria, indicating a promising approach to the management of oral infectious diseases such as dental caries and periodontal diseases. This article aims to introduce these novel approaches that have shown potential in the control of oral biofilm. These methods may be utilized in the near future to effectively promote the clinical management of oral infectious diseases and thus benefit oral health.

Molecular signalling inPseudomonas syringaepv.actinidiae

Bacterial communication is a complex phenomenon that involves a heterogeneous class of compounds acting as signal molecules and receptors that specifically recognize some of the signal molecules. This communication system occurs between bacteria of the same species, bacteria of different species and also between bacteria and eukaryotes; it affects different physiological features of the bacteria, including biofilm formation, motility and virulence. Pseudomonas syringae pv. actinidiae (Psa), the pathogen responsible of the most severe disease of kiwifruit, lacks a canonical signal molecule but harbours at least three signal receptors. This study aimed to determine i) whether Psa was able to sense its own density population using a yet-to-be characterized cell-density signalling system, ii) whether Psa was sensing the presence of other bacteria that share its ecological niche, and iii) whether any inter-kingdom interaction occurred between Psa and its hosts. Expression of genes related to Psa biofilm production, motility and virulence were analysed by qPCR in Psa samples derived from different culture conditions [high/low density, exogenous addition of synthetic acyl homoserine lactones (AHLs), co-culture with other bacteria and presence of plant-derived medium]. Biofilm formation, motility and growth in planta seemed to be only slightly influenced by Psa cell density. However, the addition of C6-OH-homoserine lactones and C8-OH-homoserine lactones increased both motility and biofilm formation. The presence of AHL-producing bacteria also modified Psa phenotype, suggesting the existence of an interspecies signalling system between kiwifruit epiphytic bacteria. In addition, we showed that crude extracts of Actinidia deliciosa were able to influence biofilm formation, Psa mobility and virulence. This phenomenon was confirmed by qPCR analysis of biofilm, motility and virulence-related gene expression. Non-host plant extracts did not affect Psa behaviour or phenotype.

A Novel Silver Bioactive Glass Elicits Antimicrobial Efficacy Against Pseudomonas aeruginosa and Staphylococcus aureus in an ex Vivo Skin Wound Biofilm Model.

Biofilm infection is now understood to be a potent contributor to the recalcitrant nature of chronic wounds. Bacterial biofilms evade the host immune response and show increased resistance to antibiotics. Along with improvements in antibiotic stewardship, effective new anti-biofilm therapies are urgently needed for effective wound management. Previous studies have shown that bioactive glass (Bg) is able to promote healing with moderate bactericidal activity. Here we tested the antimicrobial efficacy of a novel BG incorporating silver (BgAg), against both planktonic and biofilm forms of the wound-relevant bacteria Pseudomonas aeruginosa and Staphylococcus aureus. BgAg was stable, long lasting, and potently effective against planktonic bacteria in time-kill assays (6-log reduction in bacterial viability within 2 h) and in agar diffusion assays. BgAg reduced bacterial load in a physiologically relevant ex vivo porcine wound biofilm model; P. aeruginosa (2-log reduction) and S. aureus (3-log reduction). BgAg also conferred strong effects against P. aeruginosa biofilm virulence, reducing both protease activity and virulence gene expression. Co-culture biofilms appeared more resistant to BgAg, where a selective reduction in S. aureus was observed. Finally, BgAg was shown to benefit the host response to biofilm infection, directly reducing host tissue cell death. Taken together, the findings provide evidence that BgAg elicits potent antimicrobial effects against planktonic and single-species biofilms, with beneficial effects on the host tissue response. Further investigations are required to elucidate the specific consequences of BG administration on polymicrobial biofilms, and further explore the effects on host–microbe interactions.

Biofilm inhibitory efficiency of phytol in combination with cefotaxime against nosocomial pathogen Acinetobacter baumannii.

This study aimed to evaluate the antibiofilm potential of phytol and cefotaxime combinations (PCCs) against Acinetobacter baumannii and to elucidate the molecular mechanism of their antibiofilm potential through the transcriptomic approach. Phytol and cefotaxime combination(s) (PCC(s) [160μgml-1 +8μgml-1 for microbial type culture collection (MTCC) strain and 160μgml-1 +0.5μgml-1 for clinical isolate] effectively inhibited the A. baumannii biofilm formation. Additionally, light, confocal laser scanning and scanning electron microscopic analyses validated the antibiofilm potential of PCCs. Furthermore, PCCs treated A. baumannii cells showed a decreased level of hydrophobicity index compared to their respective controls. Fourier-transform infrared (FT-IR) spectra of exopolysaccharide matrix extracted from PCCs-treated A. baumannii cells showed a visible decrease in absorbance of polysaccharides, nucleic acids and protein regions compared to the spectra of untreated controls. In the blood sensitivity assay, the PCCs-treated A. baumannii plates showed reduced a number of bacterial colonies compared to their control plates. Reduced level of catalase production was also observed in the PCCs treatment compared to their controls. Transcriptomic analysis revealed the downregulation of bfmR, bap, csuA/B, ompA, pgaA, pgaC and katE biofilm virulence genes in both the A. baumannii strains on treatment with PCCs. The obtained results of this study indicate that PCCs have potent antibiofilm activity and downregulate the biofilm-related virulence genes expression in A. baumannii. To the best of our knowledge, this is the pioneering study, which shows the antibiofilm effect of PCCs against A. baumannii along with their molecular mechanism. The antibiofilm effect of PCCs could be a successful strategy for eradicating infections related to A. baumannii biofilms in nosocomial settings.

Difference in virulence and composition of a cariogenic biofilm according to substratum direction

The aim of this study was to investigate the difference in composition and virulence of Streptococcus mutans biofilms according to substratum direction. S. mutans biofilms (46-h-old) were formed on three different saliva-coated hydroxyapatite (sHA) disc direction groups: downward (discs placed in the direction of gravity), vertical (discs placed parallel to gravity direction), and upward (discs placed opposite to gravity). The 46-h-old biofilms on sHA discs in the upward direction showed the highest biofilm accumulation, colony forming unit (CFU) count, and extracellular polysaccharide (EPS) amount, followed by those in the vertical and downward directions. In the confocal laser scanning microscopy (CLSM) study, the biofilms in the upward direction also showed the highest bacterial count (live or dead cells) and EPS biovolume. Scanning electron microscopy (SEM) analysis confirmed the microbiological and biochemical results. In addition, biofilm density and acid production were higher in the upward direction than those in the other directions. Our findings suggest that substratum direction, which might be related to gravity, strongly influences the formation and virulence of cariogenic biofilms and subsequent initiation of dental caries. Collectively, the differences in the formation and virulence of cariogenic biofilms are related to the direction of tooth surface (occlusal surfaces of mandibular teeth > proximal surfaces > occlusal surfaces of maxillary teeth).

A novel Triclosan Methacrylate-based composite reduces the virulence of Streptococcus mutans biofilm.

The use of antimicrobial monomers, linked to the polymer chain of resin composites, is an interesting approach to circumvent the effects of bacteria on the dental and material surfaces. In addition, it can likely reduce the incidence of recurrent caries lesions. The aim of this study was to evaluate the effects of a novel Triclosan Methacrylate (TM) monomer, which was developed and incorporated into an experimental resin composite, on Streptococcus mutans (S. mutans) biofilms, focusing on the analyses of vicR, gtfD, gtfC, covR, and gbpB gene expression, cell viability and biofilm characteristics. The contact time between TM-composite and S. mutans down-regulated the gbpB and covR and up-regulated the gtfC gene expression, reduced cell viability and significantly decreased parameters of the structure and characteristics of S. mutans biofilm virulence. The presence of Triclosan Methacrylate monomer causes harmful effects at molecular and cellular levels in S. mutans, implying a reduction in the virulence of those microorganisms.

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English

Oral spray containing plant-derived compounds is effective against common oral pathogens

ObjectivesPlant-derived compounds are a good source of therapeutic agents and inhibitors of inflammatory process. Dental caries, periodontal diseases and candidiasis are common oral infections caused by virulent biofilms. The objectives of this study were to develop oral spray containing plant-derived compounds; α-mangostin (α-MG) and/or lawsone methyl ether (2-methoxy-1,4-naphthoquinone) (LME) and determine its antimicrobial, anti-biofilm, and anti-inflammatory activities. DesignOral spray formulations were prepared containing α-MG (5 mg/ml) and/or LME (250 μg/ml). Antimicrobial activity against Candida albicans, Streptococcus mutans, and Porphyromonas gingivalis and anti-biofilm formation activities were determined as well as cytotoxicity and anti-inflammatory effects. ResultsThe oral spray demonstrated antimicrobial activity against all three of the oral pathogens tested with stronger effects on C. albicans and S. mutans than P. gingivalis. The formulation containing α-MG (2.5 mg/ml) and LME (125 ug/ml) reduced growth of the microorganisms about 1–2 Log CFU/ml at 1–3 h and the killing effects were complete at 24 h. Based on biofilm assay, the oral spray containing both α-MG and LME showed greater inhibitory effects than those with α-MG or LME. In addition, the oral spray containing both α-MG and LME demonstrated more inhibition of nitric oxide production than α-MG alone. All the formulations were safe and demonstrated greater anti-inflammatory activity at lower concentration (<6.25 μg/ml) than at a higher concentration. ConclusionOral spray containing α-MG and/or LME is effective against common oral pathogens without significant cytotoxicity. Thus, it has the potential to prevent the infections and may serve as adjunctive treatment to conventional therapy.

The Bioactive Lipid (S)-Sebastenoic Acid Impacts Motility and Dispersion in Vibrio cholerae.

Although Gram-negative bacterial pathogens continue to impart a substantial burden on global healthcare systems, much remains to be understood about aspects of basic physiology in these organisms. In recent years, cyclic-diguanylate (c-di-GMP) has emerged as a key regulator of a number of important processes related to pathogenicity, including biofilm formation, motility and virulence. In an effort to discover chemical genetic probes for studying V. cholerae we have developed a new motility-based high-throughput screen to identify compounds that modulate c-di-GMP levels. Using this new screening platform, we tested a library of microbially-derived marine natural products extracts, leading to the discovery of the bioactive lipid (S)-sebastenoic acid. Evaluation of the effect of this new compound on bacterial motility, vpsL expression and biofilm formation implied that (S)-sebastenoic acid may alter phenotypes associated to c-di-GMP signaling in V. cholerae.

Numerical Modeling of The Quorum Sensing In a Bacterial Biofilm

In the present paper we propose a bi-dimensional non-linear reaction-diffusion model de-scribing the action of antibiotics as well as quorum sensing inhibitors agents on the virulence of bacteria biofilms. We thus approximate the system by a standard finite element scheme in space and an implicit Euler method in time. We prove the existence and uniqueness of a solution to this system and prove its convergence to a weak solution of the model we introduce. We present numerical results for simulating the quorum sensing of the very human pathogenic bacteria Pseudomenas aeruginosa.

Photoreaction of BlrP1: the role of a nonlinear photo-intensity sensor.

Blue-light-regulated phosphodiesterase 1 (BlrP1) is a blue light sensor protein that controls the hydrolysis of cyclic dimeric guanosine monophosphate, which regulates cellular motility, virulence, and formation of biofilms. In this report, the photoreaction dynamics of BlrP1 and its blue light sensor using a flavin adenine dinucleotide (BLUF) domain were investigated by the time-resolved transient grating method. Only a minor conformational change of the BlrP1-BLUF domain was observed. In contrast, a significant conformational change of the full-length BlrP1 was detected as a diffusion change with a time constant of 21 ms. Interestingly, the extent of the conformational change was concentration-dependent and the dimer form of BlrP1 was found to be the species that exhibited the conformational change. In combination with circular dichroic measurements, a quaternary structural change was determined to be the main origin of the diffusion change. Surprisingly, this conformational change was found to depend strongly on the excitation light intensity. This light-intensity-dependence indicates that the conformational change is induced by the photoexcitation of two monomer units of the dimer. The results suggest that BlrP1 is not only a photosensor but also a light intensity sensor possessing a nonlinear response.

Effect of brief periodic fluoride treatments on the virulence and composition of a cariogenic biofilm

The present study investigated the effect of periodic 1-min fluoride treatments on Streptococcus mutans biofilms and then determined the relationship between anti-biofilm activity, treatment frequency, and fluoride concentration using a linear-fitting procedure. S. mutans biofilms were periodically treated (1-min/treatment) with fluoride during biofilm formation and analyzed using microbiological methods, confocal microscopy, and real-time PCR. The results indicated that reductions in the dry weight and acidogenicity of biofilms due to periodic fluoride treatment occurred in a concentration dependent manner. The reduction in dry weight without affecting bacterial cell viability was observed mainly due to the inhibitory effect of fluoride on gtfB and gtfC gene expression, which suppresses EPS production and avoids reduction of the pH below the critical point on the tooth surface. This study suggests that brief periodic exposure to appropriate fluoride concentrations through mouthwashes and toothpastes may affect the virulence and composition of cariogenic biofilms and subsequently prevent dental caries.

Contribution of the csgA and bcsA genes to Salmonella enterica serovar Pullorum biofilm formation and virulence

ABSTRACTSalmonella biofilm formation is important to environmental stress resistance and virulence. However, the roles of the csgA and bcsA genes, which affect curli protein and cellulose production, respectively, in Salmonella enterica serovar Pullorum, are unknown. Here we constructed deletions in the csgA and bcsA genes in S. enterica serovar Pullorum strain S6702 and evaluated several aspects of biofilm formation and virulence. ΔcsgA showed decreased production of curli fimbriae, while ΔbcsA had reduced cellulose production. Both mutants had a reduced ability to form biofilms. ΔcsgA was reduced in adhesion and invasion to HeLa cells and exhibited decreased intracellular proliferation in HD11 macrophages. ΔbcsA exhibited increased proliferation in HD11 cells and replicated better in chicken spleens, as compared to the wild-type strain. ΔcsgA virulence was attenuated in assays involving oral challenge of one-day-old chickens.