Multispecies Biofilm Research Articles

Amyloid hexapeptide prevent dental caries by antibiofilm formation

ObjectivesBiofilm formed by cariogenic microbes is the direct cause of dental caries, therefore, prevention of dental caries should be anti-biofilm-based. Previously, we found the amyloid hexapeptides efficiently inhibited biofilm formation by aggregating into amyloid fibrils agglutinating microbes. This study aimed to select the most stable amyloid hexapeptide GIDLKI (GI6) and study its anti-caries effect. MethodsBiofilms of multi-species bacteria, derived from mixed saliva, were cultured to evaluate the anti-biofilm formation effect of GI6. And then, the primary cariogenic bacterium Streptococcus mutans (S.mutans) was cultured in BHI with various pH, gradient concentrations of sucrose, glucose, and calcium ions to evaluate the anti-biofilm formation effects of GI6. Then models of human enamel block caries and twenty male SPF-SD rat caries induced by S. mutans biofilm were constructed, and confocal laser scanning microscopy, scanning electron microscopy, and micro-computed tomography were applied to investigate the anti-biofilm formation, anti-caries effects and use safety of GI6. ResultsGI6 could inhibit the multi-species bacteria biofilm formation and remained effective in anti-biofilm activity against S. mutans in environments closely related to caries. GI6 suppressed S. mutans biofilm formation and thus prevented or alleviated the development of caries in human tooth blocks and rat teeth. GI6 did not affect the intestinal flora, serum biochemical parameters, and the pathological changes of various organs. ConclusionsAmyloid hexapeptides, including but not limited to GI6, are novel effective anti-caries agents that can be used to prevent dental caries safely. Clinical significanceThis study explored the anti-biofilm formation and anti-caries effect of GI6 in vitro, highlighting the anti-biofilm formation therapy for dental caries and setting a foundation for the practical application of GI6 for the treatment of dental caries.

Effects of caffeic acid phenethyl ester against multi-species cariogenic biofilms.

Dental caries is a biofilm-related disease, widely perceived to be caused by oral ecological imbalance when cariogenic/aciduric bacteria obtain an ecological advantage. Compared with planktonic bacteria, dental plaques are difficult to remove under extracellular polymeric substance protection. In this study, the effect of caffeic acid phenethyl ester (CAPE) on a preformed cariogenic multi-species biofilm was evaluated, which was comprised of cariogenic bacteria (Streptococcus mutans), commensal bacteria (Streptococcus gordonii), and a pioneer colonizer (Actinomyces naeslundii). Our result revealed that treatment with 0.08mg/mL CAPE reduced live S. mutans in the preformed multi-species biofilm while not significantly changing the quantification of live S. gordonii. CAPE significantly reduced the production of lactic acid, extracellular polysaccharide, and extracellular DNA and made the biofilm looser. Moreover, CAPE could promote the H2O2 production of S. gordonii and inhibit the expression of SMU.150 encoding mutacin to modulate the interaction among species in biofilms. Overall, our results suggested that CAPE could inhibit the cariogenic properties and change the microbial composition of the multi-species biofilms, indicating its application potential in dental caries prevention and management.

Effect of different disinfection protocols in bacterial viability of an intraradicular biofilm formed in situ.

The present study aimed to evaluate bacterial viability after the use of different disinfection protocols in root canals infected with a multispecies biofilm (MB) formed in situ. Palatal roots with a single canal were obtained from extracted maxillary molars and sterilized before being inserted into the mouth. The roots were contaminated with a MB in an intraoral appliance worn by ten volunteers. All volunteers wore six roots simultaneously in two intraoral devices for 21 days. One root from each volunteer was assigned to each group (n=10): PUI - passive ultrasonic irrigation; EC - Easy Clean; XPF - XP-endo Finisher; aPDT - antimicrobial photodynamic therapy; CI - conventional irrigation; and NC - negative control. The samples were evaluated under confocal laser scanning microscopy. The percentage of viable cells (VC) was calculated over the total percentage of MB biovolume. Data were statistically analyzed (α=5%). The cell viability in the entire root canal or for each third was compared between groups (Kruskal-Wallis test, Dunn post-hoc test) and for the same group (Friedman test, Dunn post-hoc test). Disinfection protocols were not significantly different from each other (P>.05). Samples in EC, PUI, and aPDT had lower cell viability than in NC (P<.05). In the coronal third of samples in the EC, XPF, PUI and aPDT, the percentage of VC biovolume was lower than in the NC (P<.05). The percentage of VC in EC samples was lower in the coronal and middle thirds than in the apical third (P<.05). EC, PUI and aPDT had significant effects on cell viability in intraradicular multispecies biofilm formed in situ when compared with untreated samples.

Analysis of antimicrobial efficacy of sodium hypochlorite and ozonated water against biofilm in oval canals.

This in vitro study compared the antimicrobial efficacy of 2.5% sodium hypochlorite (NaOCl) and 8 µg/mL ozonated water agitated by passive ultrasonic irrigation (PUI) or PUI combined with EndoActivator (EA) against mature multispecies biofilm. One hundred and five oval-shaped mandibular premolars were instrumented, sterilized, and inoculated with Enterococcus faecalis, Candida albicans, and Staphylococcus aureus, divided into: control group - saline; O3 group - ozonated water; O3 PUI group - ozonated water with PUI agitation; O3 PUI+EA group - ozonated water with PUI+EA agitation; NaOCl group - NaOCl; NaOCl PUI group - NaOCl with PUI agitation; and NaOCl PUI+EA group - NaOCl with PUI+EA agitation. Microbiological samples were collected before (S1) and after (S2) the disinfection procedures and the data were statistically analyzed using the Kruskal-Wallis test. In the culture method, there was significant disinfection in the O3 PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). The combination of NaOCl with PUI+EA reduced microbial counts to zero (p˂0.05). In the qPCR method, there was a significant reduction in the total count of viable microorganisms in the O3 PUI, O3 PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). It can be concluded that 2.5% NaOCl with and without agitation, as well as 8 µg/mL ozonated water with its action enhanced by the agitation techniques, were effective in root canal disinfection, and their antimicrobial efficacy is related to the microorganisms present in the biofilm.

In vitro evaluation of membranes for regenerative procedures against oral bacteria.

The current literature on guided bone regeneration (GBR) and guided tissue regeneration (GTR) membrane contamination reports that the physicochemical characteristics of these biomaterials might influence affinity to bacteria, which appears to be a major drawback for the clinical outcome of the regenerative procedures. Thus, this study aimed to evaluate, in vitro, a multispecies biofilm adherence and passage of bacteria through different types of commercially available membranes for GTR/GBR. Four types of membranes were tested (n=12): LC) Lumina Coat®; JS) Jason®; BG) Biogide®; and LP) Lumina PTFE®. Aluminum foil (AL) simulated an impermeable barrier and was used as the control. The membranes were adapted to specific apparatus and challenged with a mixed bacterial culture composed of A. actinomycetemcomitans b, S. mutans, S. mitis, and A. israelii. After 2 h or 7 days, bacterial adhesion and passage of bacteria were evaluated through CFU counting, which was analyzed by two-way ANOVA e post hoc Tukey, at a 5% significance level. Representative areas of two membranes of each group were analyzed through scanning electron microscopy (SEM) to assess the morphology and organization of the biofilm over the membrane fibers. LC and LP presented similar values of adhered bacterial cells (p > 0.05), significantly inferior when compared to the other groups, in both time points (p < 0.05). All the tested groups were permeable to bacterial cells, with no significant difference between the trial period of 2 h and 7 days (p > 0.05). SEM analyses demonstrated that adhered bacteria number increased throughout the time points (2 h < 7 days). Commercially available biological membranes demonstrated intense bacterial adherence and passage of bacteria, which increased throughout the trial period.

In vitro and in situ caries-preventive effect of a new combined fluoride and calcium experimental nanocomposite solution.

To assess the in vitro and in situ effect of experimental combined fluoride and calcium nanocomposite solutions on dental caries prevention. Nanocompound mesoporous silica (MS) with calcium (Ca) and sodium fluoride (NaF) - (MSCaNaF); MS with NaF (MSNaF), NaF solution (positive control), and deionized water (negative control - CG) were studied. The specimens (n=130) were submitted in vitro to a multispecies biofilm in the presence of 2% sucrose. After 24 h and 48 h, the culture medium pH, the percent of surface mineral loss (%SML), and lesion depth (ΔZ) were analyzed. In the in situ study, 10 volunteers participated in four phases of 7-days each. The products were applied on the specimens (n=240) before 20% sucrose solution drips. The polysaccharides (SEPS and IEPS), %SML and roughness (Sa) were evaluated. There was an in vitro decrease in pH values in 24h and 48h, compared to baseline. The MSCaNaF and MSNaF groups obtained lower values of %SML and ΔZ (p < 0.05) than CG and NaF after 24h and were similar to NaF after 48h (p<0.05). In situ results showed similar SEPS and IEPS among all groups after 48h. An after 7-days, the nanocomposites had similar values (p>0.05), while NaF was similar to CG (p>0.05). After 48h, the MSCaNaF and MSNaF reduced the %SML (p<0.05). After 7-days, both experimental nanocomposites were similar to NaF (p>0.05). Regarding Sa, MSCaNaF was better than NaF for both periods (p<0.05). The nanocomposites controlled the in vitro and in situ enamel demineralization, mainly in the initial periods.

High Disinfectant Tolerance in Pseudomonas spp. Biofilm Aids the Survival of Listeria monocytogenes.

Pseudomonas spp. are the most commonly found bacteria in food-processing environments due to properties such as a high growth rate at low temperatures, a high tolerance of antimicrobial agents, and biofilm formation. In this study, a set of Pseudomonas isolates originating from cleaned and disinfected surfaces in a salmon processing facility were screened for biofilm formation at 12 °C. A high variation in biofilm formation between the isolates was observed. Selected isolates, in both planktonic and biofilm states, were tested for resistance/tolerance to a commonly used disinfectant (peracetic acid-based) and antibiotic florfenicol. Most isolates showed a much higher tolerance in the biofilm state than in the planktonic state. In a multi-species biofilm experiment with five Pseudomonas strains with and without a Listeria monocytogenes strain, the Pseudomonas biofilm appeared to aid the survival of L. monocytogenes cells after disinfection, underscoring the importance of controlling the bacterial load in food-processing environments.

Phylogenetic Constitution and Survival of Microbial Biofilms Formed on the Surface of Polyethylene Composites Protected with Polyguanidine Biocides

A series of biocide-containing polyethylene composites were obtained using novel guanidine-containing copolymers immobilized on an inert mineral carrier. Multispecies microbial communities were isolated from the surface of polyethylene samples either incubated or found in the environment, and their taxonomic composition was determined. Biofilms reconstructed using microorganisms obtained from different ecotopes were shown to intensively foul polyethylene surfaces. The presence of polyguanidine biocide suppressed the growth and survival of both binary and multispecies biofilms, with a cumulative effect during long-term incubation. When microorganisms were co-cultivated in binary biofilms, the phenomenon of a decrease in biocide effectiveness was demonstrated. This protective effect is potentially based on cooperative interactions inside the binary biofilm community. Scanning electron microscopy showed a pronounced difference in the architecture of reconstructed biofilms incubated in the presence of biocide in comparison to control samples, where biocide suppressed the formation of dense and well-organized three-dimensional structures. Biofilm disruption by immobilized biocides occurred primarily during the later stages of biofilm formation, probably caused by polycation interaction with their negatively charged extracellular components.

Discovery of phosphorylated lantibiotics with proimmune activity that regulate the oral microbiome

Lantibiotics are ribosomally synthesized and posttranslationally modified peptides (RiPPs) that are produced by bacteria. Interest in this group of natural products is increasing rapidly as alternatives to conventional antibiotics. Some human microbiome-derived commensals produce lantibiotics to impair pathogens' colonization and promote healthy microbiomes. Streptococcus salivarius is one of the first commensal microbes to colonize the human oral cavity and gastrointestinal tract, and its biosynthesis of RiPPs, called salivaricins, has been shown to inhibit the growth of oral pathogens. Herein, we report on a phosphorylated class of three related RiPPs, collectively referred to as salivaricin 10, that exhibit proimmune activity and targeted antimicrobial properties against known oral pathogens and multispecies biofilms. Strikingly, the immunomodulatory activities observed include upregulation of neutrophil-mediated phagocytosis, promotion of antiinflammatory M2 macrophage polarization, and stimulation of neutrophil chemotaxis-these activities have been attributed to the phosphorylation site identified on the N-terminal region of the peptides. Salivaricin 10 peptides were determined to be produced by S. salivarius strains found in healthy human subjects, and their dual bactericidal/antibiofilm and immunoregulatory activity may provide new means to effectively target infectious pathogens while maintaining important oral microbiota.

Effect of cetrimonium carrier micelles on bacterial membranes and extracellular DNA, an in silico study

Microorganisms do not live as dispersed single cells but rather they form aggregates with extracellular polymeric substances at interfaces. Biofilms are considered efficient life forms because they shield bacteria from biocides and collect dilute nutrients. This is a big concern in industry since the microorganisms can colonize a wide range of surfaces, accelerating material deterioration, colonizing medical devices, contaminating ultrapure drinking water, increasing energy costs and creating focus of infection. Conventional biocides that target a specific component of the bacteria are not effective in the presence of biofilms. Efficient biofilm inhibitors are based on a multitarget approach interacting with the bacteria and the biofilm matrix. Their rationale design requires a thorough understanding of inhibitory mechanisms that are still largely lacking today. Herein we uncover via molecular modelling the inhibition mechanism of cetrimonium 4-OH cinnamate (CTA-4OHcinn). Simulations show that CTA-4OH micelles can disrupt symmetric and asymmetric bilayers, representative of inner and outer bacterial membranes, following three stages: adsorption, assimilation, and defect formation. The main driving force for micellar attack is electrostatic interactions. In addition to disrupting the bilayers, the micelles work as carriers facilitating the trapping of 4OH cinnamate anions within the bilayer upper leaflet and overcoming electrostatic repulsion. The micelles also interact with extracellular DNA (e-DNA), which is one of the main components of biofilms. It is observed that CTA-4OHcinn forms spherical micelles on the DNA backbone; which hinders their ability to pack. This is demonstrated by modelling the DNA along the hbb histone-like protein, showing that in the presence of CTA-4OHcinn, DNA does not pack properly around hbb. The abilities of CTA-4OHcinn to cause cell death through membrane disruption and to disperse a mature, multi-species biofilm are also confirmed experimentally.

Inhibitory effects of piperine and black pepper essential oil on multispecies biofilm formation by Listeria monocytogenes, Salmonella Typhimurium, and Pseudomonasaeruginosa

This study aimed to evaluate the antimicrobial and antibiofilm activities of black pepper essential oil (BPEO) and piperine and the effect of piperine on gene expression in a multispecies biofilm composed of Listeria monocytogenes, Salmonella Typhimurium, and Pseudomonas aeruginosa on a polypropylene surface. The minimal inhibitory concentrations of BPEO and piperine were 100 and 25 mg/mL, respectively, against this consortium of microorganisms. Sessile cell counts were 5.35–7.35 log CFU/cm2 and varied over time. The total population eradicated in the biofilm ranged from 78.9% (5.88 log CFU/cm2) to 99.8% (4.16 log CFU/cm2). Evaluation of biofilm-related gene expression showed upregulation of the L. monocytogenes genes agrC (24 and 72 h), agrD (72 h), and prfA (72 h) and downregulation of all evaluated S. Typhimurium and P. aeruginosa genes (24 and 72 h) in the untreated control biofilm. The addition of piperine resulted in upregulation of the L. monocytogenes genes agrB (24 and 72 h), agrC (72 h), agrD (24 and 72 h), and prfA (24 h); the S. Typhimurium genes agfA (24 and 72 h), adrA (24 and 72 h), and csgD (72 h); and all evaluated P. aeruginosa genes (24 and 72 h). Piperine more effectively controlled the multispecies biofilm.

A review of mechanism analysis methods in multi-species biofilm of foodborne pathogens.

Biofilms are an aggregation of microorganisms that have high resistance to antimicrobial agents. In the food industry, it has been widely studied that foodborne pathogens on both food surfaces and food-contact surfaces can form biofilms thereby threatening the safety of the food. In the natural environment, multi-species biofilms formed by more than two different microorganisms are abundant. In addition, the resistance of multi-species biofilms to antimicrobial agents is higher than that of mono-species biofilms. Therefore, studies to elucidate the mechanisms of multi-species biofilms formed by foodborne pathogens are still required in the food industry. In this review paper, we summarized the novel analytical methods studied to evaluate the mechanisms of multi-species biofilms formed by foodborne pathogens by dividing them into four categories: spatial distribution, bacterial interaction, extracellular polymeric substance production and quorum sensing analytical methods.

Effect of outdoor pipe materials and community-intrinsic properties on biofilm formation and chlorine resistance: Black sheep or team leader

Community-intrinsic properties affect the function and composition of microbial community, and understanding of the microbial community-intrinsic properties in different pipe materials of drinking water distribution systems can help to prevent microbial pollution more reasonably and efficiently. This study investigated the biofilm formation and chlorine resistance of multi-species biofilms in two representative outdoor pipe materials (polyethylene (PE) and cast iron pipe). The results showed that cast iron pipes have higher biofilm biomass but lower biofilm activity than PE pipes. The different growth environments of the two pipe materials resulted in diverse microbial community-intrinsic properties and affected multi-species biofilm formation. The microbial communities without Sphingomonas or Acidovorax in cast iron pipe exhibited higher biofilm biomass, when compared with those in PE pipe. In contrast, the microbial community without Bacillus in PE pipe presented higher biofilm biomass, when compared with those in cast iron pipe. Extracellular polymeric substance (EPS) and corrosion products protected bacteria from chlorination. Although community-intrinsic properties helped bacteria resist chlorination, the same microbial community exhibited diverse chlorine resistance in different pipe materials owing to different community-intrinsic properties. While the chlorine resistance of microbial community without Bacillus was low in PE pipe, that in cast iron pipe was better owing to high corrosion degree. Multi-species biofilm without Sphingomonas or Acidovorax presented higher biofilm biomass, but lower EPS content and corrosion products in cast iron pipe, resulting in low chlorine resistance. In summary, this study revealed that outdoor pipe materials could affect microbial community-intrinsic properties, which in turn could influence biofilm characteristics, and provided unique insights for ensuring drinking water safety.

The effect of synbiotic-fluoride therapy on multi-species biofilm

ObjectivesThe study objective was to examine the effect of synbiotic-fluoride (SF) therapy within a multi-species cariogenic biofilm model system comprising of S. mutans, S. sanguinis, and S. gordonii. MethodsThe SF therapy was prepared using 2% L-arginine (Arg), 0.2% NaF and probiotic L. rhamnosus GG (LRG). The 8 treatment groups were: Group 1: No treatment, Group 2: 2% Arg, Group 3: 0.2% NaF, Group 4: LRG, Group 5: 2% Arg+0.2% NaF, Group 6: 2% Arg+LRG, Group 7: 0.2% NaF+LRG, and Group 8: SF therapy (2% Arg+0.2% NaF +LRG). Multi-species biofilm model over 96 h comprising Streptococcus mutans, Streptococcus sanguinis, and Streptococcus gordonii was utilized. The biofilms received cariogenic challenge and SF therapy 2 × /day. The extracellular matrix components were analyzed for carbohydrates, proteins, and extra-cellular DNA (eDNA). The live/dead cells were imaged and quantified using confocal microscopy. The viable/dead bacterial concentrations were estimated using propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR). The gene expressions for gtfB, sagP, arcA, argG, and argH were measured using real-time reverse transcriptase qPCR. ResultsCarbohydrates and protein content with SF therapy were higher than non-LRG containing groups, while eDNA content was lower than other groups (p<0.05). Live bacterial proportions determined using confocal imaging with SF therapy were the lowest (p<0.05). The 2% Arg+LRG and SF therapy showed higher viable L. rhamnosus GG than 0.2% NaF+LRG (p<0.05). The dead S. mutans with SF therapy were higher than the other groups (p<0.05) with no difference from 2% Arg+0.2% NaF and 2% Arg+LRG (p>0.05). The SF therapy significantly downregulates gtfB and upregulates sagP, arcA, argG, argH gene expression (p<0.05). ConclusionSynbiotic-fluoride therapy effectuates multi-fold changes in the multi-species biofilm matrix and cellular components leading to superior ecological homeostasis than its individual contents, prebiotics (arginine), probiotic (L. rhamnosus GG), and fluorides (NaF). Clinical significanceThe ecological-based synbiotic-fluoride caries-preventive therapy aids in maintaining biofilm homeostasis to preempt/restore dysbiosis thereby sustaining dynamic-diverse health-associated microbial stability significant as a preventive regimen for high caries-risk patients.

Streptococcus salivarius as an Important Factor in Dental Biofilm Homeostasis: Influence on Streptococcus mutans and Aggregatibacter actinomycetemcomitans in Mixed Biofilm.

A disturbed balance within the dental biofilm can result in the dominance of cariogenic and periodontopathogenic species and disease development. Due to the failure of pharmacological treatment of biofilm infection, a preventive approach to promoting healthy oral microbiota is necessary. This study analyzed the influence of Streptococcus salivarius K12 on the development of a multispecies biofilm composed of Streptococcus mutans, S. oralis and Aggregatibacter actinomycetemcomitans. Four different materials were used: hydroxyapatite, dentin and two dense polytetrafluoroethylene (d-PTFE) membranes. Total bacteria, individual species and their proportions in the mixed biofilm were quantified. A qualitative analysis of the mixed biofilm was performed using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The results showed that in the presence of S. salivarius K 12 in the initial stage of biofilm development, the proportion of S. mutans was reduced, which resulted in the inhibition of microcolony development and the complex three-dimensional structure of the biofilm. In the mature biofilm, a significantly lower proportion of the periodontopathogenic species A. actinomycetemcomitans was found in the salivarius biofilm. Our results show that S. salivarius K 12 can inhibit the growth of pathogens in the dental biofilm and help maintain the physiological balance in the oral microbiome.

Synergistic interactions in multispecies biofilm combinations of bacterial isolates recovered from diverse food processing industries.

Most biofilms within the food industry are formed by multiple bacterial species which co-exist on surfaces as a result of interspecies interactions. These ecological interactions often make these communities tolerant against antimicrobials. Our previous work led to the identification of a large number (327) of highly diverse bacterial species on food contact surfaces of the dairy, meat, and egg industries after routine cleaning and disinfection (C&D) regimes. In the current study, biofilm-forming ability of 92 bacterial strains belonging to 26 genera and 42 species was assessed and synergistic interactions in biofilm formation were investigated by coculturing species in all possible four-species combinations. Out of the total 455 four-species biofilm combinations, greater biofilm mass production, compared to the sum of biofilm masses of individual species in monoculture, was observed in 34 combinations. Around half of the combinations showed synergy in biofilm mass > 1.5-fold and most of the combinations belonged to dairy strains. The highest synergy (3.13-fold) was shown by a combination of dairy strains comprising Stenotrophomonas rhizophila, Bacillus licheniformis, Microbacterium lacticum, and Calidifontibacter indicus. The observed synergy in mixed biofilms turned out to be strain-specific rather than species-dependent. All biofilm combinations showing remarkable synergy appeared to have certain common species in all combinations which shows there are keystone industry-specific bacterial species which stimulate synergy or antagonism and this may have implication for biofilm control in the concerned food industries.

Antimicrobial Photodynamic Inactivation: An Alternative for Group B Streptococcus Vaginal Colonization in a Murine Experimental Model.

Streptococcus agalactiae, referred to as Group B Streptococcus (GBS), is a prominent bacterium causing life-threatening neonatal infections. Although antibiotics are efficient against GBS, growing antibiotic resistance forces the search for alternative treatments and/or prevention approaches. Antimicrobial photodynamic inactivation (aPDI) appears to be a potent alternative non-antibiotic strategy against GBS. The effect of rose bengal aPDI on various GBS serotypes, Lactobacillus species, human eukaryotic cell lines and microbial vaginal flora composition was evaluated. RB-mediated aPDI was evidenced to exert high bactericidal efficacy towards S. agalactiae in vitro (>4 log10 units of viability reduction for planktonic and >2 log10 units for multispecies biofilm culture) and in vivo (ca. 2 log10 units of viability reduction in mice vaginal GBS colonization model) in microbiological and metagenomic analyses. At the same time, RB-mediated aPDI was evidenced to be not mutagenic and safe for human vaginal cells, as well as capable of maintaining the balance and viability of vaginal microbial flora. aPDI can efficiently kill GBS and serve as an alternative approach against GBS vaginal colonization and/or infections.

Core Species Derived from Multispecies Interactions Facilitate the Immobilization of Cadmium

Microbial consortia have opened new avenues for heavy-metal remediation. However, the limited understanding of the overall effect of interspecific interactions on remediation efficacy hinders its application. Here, the effects of multispecies growth and biofilm formation on Cd immobilization were explored from direct and multiple interactions through random combinations of two or three rhizosphere bacteria. In monocultures, Cd stress resulted in an average decrease in planktonic biomass of 26%, but through cooperation, the decrease was attenuated in dual (21%) and triple cultures (13%), possibly involving an increase in surface polysaccharides. More than 65% of the co-cultures exhibited induction of biofilm formation under Cd stress, which further enhanced the role of biofilms in Cd immobilization. Notably, excellent biofilm-forming ability or extensive social induction makes Pseudomonas putida and Brevundimonas diminuta stand out in multispecies biofilm formation and Cd immobilization. These two core species significantly increase the colonization of soil microorganisms on rice roots compared to the control, resulting in a 40% decrease in Cd uptake by rice. Our study enhances the understanding of bacterial interactions under Cd stress and provides a novel strategy for adjusting beneficial soil consortia for heavy-metal remediation.

Electrochemical decontamination of titanium dental implants. An in vitro biofilm model study.

The objective of this study is to study the effect of electrochemical treatment on biofilms developed on titanium dental implants, using a six-species in vitro model simulating subgingival oral biofilms. Direct electrical current (DC) of 0.75 V, 1.5V, and 3V (anodic polarization, oxidation processes) and of -0.75 V, -1.5V, and -3V (cathodic polarization, reduction processes) was applied between the working and the reference electrodes for 5min on titanium dental implants, which have been previously inoculated with a multispecies biofilm. This electrical application consisted of a three-electrode system where the implant was the working electrode, a platinum mesh was the counter electrode, and an Ag/AgCl electrode was the reference. The effect of the electrical application on the biofilm structure and bacterial composition was evaluated by scanning electron microscopy and quantitative polymerase chain reaction. A generalized linear model was applied to study the bactericidal effect of the proposed treatment. The electrochemical construct at 3V and -3V settings significantly reduced total bacterial counts (p < .05) from 3.15 × 106 to 1.85 × 105 and 2.92 × 104 live bacteria/mL, respectively. Fusobacterium nucleatum was the most affected species in terms of reduction in concentration. The 0.75 V and -0.75 V treatments had no effect on the biofilm. Electrochemical treatments had a bactericidal effect on this multispecies subgingival in vitro biofilm model, being the reduction more effective than the oxidative treatment.

Biofilm formation on dental implants with a hybrid surface microtopography: An in vitro study in a validated multispecies dynamic biofilm model.

The objective of this study is to qualitatively and quantitatively evaluate biofilm formation on hybrid titanium implants (HS), with moderately rough and turned surface topographies. A validated dynamic in vitro multispecies biofilm model, based on bacterial growth under flow and shear conditions resembling the oral cavity, was used to evaluate biofilm formation on the tested implant surfaces. Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) were used to compare the biofilm structure and microbial biomass deposited on either the moderately rough or the turned surface of HS. Quantitative polymerase chain reaction (qPCR) was used to evaluate the total bacterial counts and counts of each specific bacterium in biofilms formed on implants with either the moderately rough or the turned surfaces, as in the hybrid titanium implants, after 24, 48 and 72 h. A general linear model was applied to compare the CLSM and qPCR results between the tested implant surfaces. A significantly higher bacterial biomass grew on the moderately rough implant surfaces, compared to the turned surface area of HS implants (p < .05), at all incubation times, as evidenced with both CLSM and SEM. qPCR analysis also demonstrated an important increase in the total and specific bacterial counts in moderately rough surface implants at the three incubation times. Implant surface topography (moderately rough versus turned) significantly influenced in vitro biofilm formation in terms of biofilm structure, bacterial biomass and quantity of the specific species selected for the model used.