Bacterial Composition Of Biofilms Research Articles

Antimicrobial Activity of Methylene Blue Associated with Photodynamic Therapy: In Vitro Study in Multi-Species Oral Biofilm.

The control of infectious diseases caused by biofilms is a continuing challenge for researchers due to the complexity of their microbial structures and therapeutic implications. Photodynamic therapy as an adjunctive anti-infective treatment has been described as a possible valid approach but has not been tested in polymicrobial biofilm models. This study evaluated the effect of photodynamic therapy in vitro with methylene blue (MB) 0.01% and red LEDs (λ = 660 nm, power density ≈ 330 mW/cm2, 2 mm distance from culture) on the metabolic activity and composition of a multispecies subgingival biofilm. Test Groups LED and MB + LED showed a more significant reduction in metabolic activity than the non-LED application group (~50 and 55%, respectively). Groups LED and MB equally affected (more than 80%) the total bacterial count in biofilms. No differences were noted in the bacterial biofilm composition between the groups. In vitro LED alone or the MB + LED combination reduced the metabolic activity of bacteria in polymicrobial biofilms and the total subgingival biofilm count.

Biological and Synthetic Surfactants Increase Class I Integron Prevalence in Ex Situ Biofilms.

The role of biocides in the spread of antimicrobial resistance (AMR) has been addressed but only a few studies focus on the impact of surfactants on microbial diversity and AMR, although they are common constituents of cleaners, disinfectants, and personal care products and are thus released into the environment in large quantities. In this study, we used a static ex situ biofilm model to examine the development of four biofilms exposed to surfactants and analyzed the biofilms for their prevalence of class I integrons as a proxy for the overall abundance of AMR in a sample. We furthermore determined the shift in bacterial community composition by high-resolution melt analysis and 16S ribosomal RNA (16S rRNA) gene sequencing. Depending on the initial intrinsic prevalence of class I integrons in the respective ex situ biofilm, benzalkonium chloride, alkylbenzene sulfonate, and cocamidopropyl betaine increased its prevalence by up to 6.5× on average. For fatty alcohol ethoxylate and the biosurfactants sophorolipid and rhamnolipid, the mean increase did not exceed 2.5-fold. Across all surfactants, the increase in class I integrons was accompanied by a shift in bacterial community composition. Especially benzalkonium chloride, cocamidopropyl betaine, and alkylbenzene sulfonate changed the communities, while fatty alcohol ethoxylate, sophorolipid, and rhamnolipid had a lower effect on the bacterial biofilm composition.

The Evolving Microbiome of Dental Caries.

Dental caries is a significant oral and public health problem worldwide, especially in low-income populations. The risk of dental caries increases with frequent intake of dietary carbohydrates, including sugars, leading to increased acidity and disruption of the symbiotic diverse and complex microbial community of health. Excess acid production leads to a dysbiotic shift in the bacterial biofilm composition, demineralization of tooth structure, and cavities. Highly acidic and acid-tolerant species associated with caries include Streptococcus mutans, Lactobacillus, Actinomyces, Bifidobacterium, and Scardovia species. The differences in microbiotas depend on tooth site, extent of carious lesions, and rate of disease progression. Metagenomics and metatranscriptomics not only reveal the structure and genetic potential of the caries-associated microbiome, but, more importantly, capture the genetic makeup of the metabolically active microbiome in lesion sites. Due to its multifactorial nature, caries has been difficult to prevent. The use of topical fluoride has had a significant impact on reducing caries in clinical settings, but the approach is costly; the results are less sustainable for high-caries-risk individuals, especially children. Developing treatment regimens that specifically target S. mutans and other acidogenic bacteria, such as using nanoparticles, show promise in altering the cariogenic microbiome, thereby combatting the disease.

Microbial dynamics in Manaus river, Brazil

Rivers, which are vital ecosystems that support a variety of life forms and provide many ecosystem services, have been degraded by human activities, such as urbanization, pollution, and overexploitation. Habitat restoration projects have been implemented in different parts of the world to restore the ecological health and functioning of degraded rivers. The microbial community that forms biofilms on various substrates is one of the key components of river ecosystems. Organic matter is decomposed and carbon dioxide is produced by these biofilms, which affects the oxygen levels and nutrient cycles in the water. However, the effects of habitat restoration on the biofilm microbial community are not well understood. High-throughput DNA sequencing was used in this study to compare the bacterial composition of biofilms in three types of rivers: degraded urban rivers, urban rivers with restored habitats, and forested rivers that serve as reference conditions. The following questions were aimed to be answered: (i) How is the biofilm bacterial community composition affected by habitat restoration? (ii) How does bacterial diversity change in restored rivers compared to degraded and reference rivers? (iii) What are the environmental factors that influence the biofilm bacterial community composition? It was found that water quality was improved by habitat restoration by increasing the dissolved oxygen and reducing the organic pollutants in urban rivers. A decrease in bacterial diversity and a shift in the biofilm bacterial community structure towards that of the reference forested rivers were accompanied by this. This suggests that the biofilm microbial community can be altered by habitat restoration and its role in processing organic pollutants can be enhanced. However, it was also found that there was still a significant difference in the biofilm bacterial community between the restored and reference rivers, indicating that more time and effort are needed to achieve a similar ecological quality as the natural forested rivers.

High resolution ITS amplicon melting analysis as a tool to analyse microbial communities of household biofilms in ex-situ models

Biofilms are the most common growth types of microorganisms. These complex communities usually consist of different species and are embedded in an extracellular matrix containing polymers, proteins and DNA. This matrix offers protection against different (a)biotic environmental factors and generally increases resistances. Higher resistances against antibiotics are one of the main reasons why biofilms are often associated with healthcare settings. Nevertheless, they are also found in domestic settings, mostly in humid places with abundant nutrients like dishwashers or washing machines. Biofilms in these areas show individual compositions and are influenced for example by temperature, frequency of use or the age of the device.In this study, we introduce a model for the ex-situ cultivation of domestic biofilms from household appliances. Furthermore, we tested the ability of high resolution melting analysis (HRMA) as a tool for analysing these biofilms. Our goal was to maintain a high amount of complexity in the ex-situ biofilms that is characterized by the melting behavior of the contained DNA. Dishwasher and washing machine biofilms were sampled in private households and cultivated for 10 d. After DNA extraction, 16S rDNA was sequenced and melting behavior of the bacterial Internal Transcribed Spacer (ITS) region was analysed. Additionally, testing for independence of continuous new sampling, storage of cultivated biofilms in glycerol stocks and following recultivation of them was done up to three times.Our results show that a high level of complexity could be maintained in the ex-situ biofilms after 10 d of cultivation, although in general the bacterial diversity slightly decreased compared to the original biofilm in most cases. Recultivation of a similar biofilm from glycerol stocks was possible as well with some impact by various factors. Differences in the bacterial composition of biofilms could clearly made visible by HRMA although it was not possible to match peaks to a specific phylogenetic group. Still, HRMA proved to be a less costly and time consuming alternative to sequencing for the characterization of biofilms.

Investigating the chemical pathway to the formation of a single biofilm using infrared spectroscopy

Diagnosing biofilm infections has remained a constant challenge for the last 50 years. Existing diagnostic methods struggle to identify the biofilm phenotype. Moreover, most methods of biofilm analysis destroy the biofilm making the resultant data interpretation difficult. In this study we introduce Fourier Transform Infra-Red (FTIR) spectroscopy as a label-free, non-destructive approach to monitoring biofilm progression. We have utilised FTIR in a novel application to evaluate the chemical composition of bacterial biofilms without disrupting the biofilm architecture. S. epidermidis (RP62A) was grown onto calcium fluoride slides for periods of 30 min–96 h, before semi-drying samples for analysis. We report the discovery of a chemical marker to distinguish between planktonic and biofilm samples. The appearance of new proteins in biofilm samples of varying maturity is exemplified in the spectroscopic data, highlighting the potential of FTIR for identifying the presence and developmental stage of a single biofilm.

Are late hernia mesh complications linked to Staphylococci biofilms?

PurposeThe purpose of this study was to investigate the link between bacterial biofilms and negative outcomes of hernia repair surgery. As biofilms are known to play a role in mesh-related infections, we investigated the presence of biofilms on hernia meshes, which had to be explanted due to mesh failure without showing signs of bacterial infection.MethodsIn this retrospective observational study, 20 paraffin-embedded tissue sections from explanted groin hernia meshes were analysed. Meshes have been removed due to chronic pain, hernia recurrence or mesh shrinkage. The presence and bacterial composition of biofilms were determined. First, specimens were stained with fluorescence in situ hybridisation (FISH) probes, specific for Staphylococcus aureus and coagulase-negative staphylococci, and visualised by confocal laser scanning microscopy. Second, DNA was extracted from tissue and identified by S. aureus and S. epidermidis specific PCR.ResultsConfocal microscopy showed evidence of bacterial biofilms on meshes in 15/20 (75.0%) samples, of which 3 were positive for S. aureus, 3 for coagulase-negative staphylococci and 9 for both species. PCR analysis identified biofilms in 17/20 (85.0%) samples, of which 4 were positive for S. aureus, 4 for S. epidermidis and 9 for both species. Combined results from FISH/microscopy and PCR identified staphylococci biofilms in 19/20 (95.0%) mesh samples. Only 1 (5.0%) mesh sample was negative for bacterial biofilm by both techniques.ConclusionResults suggest that staphylococci biofilms may be associated with hernia repair failure. A silent, undetected biofilm infection could contribute to mesh complications, chronic pain and exacerbation of disease.

Acylase enzymes disrupting quorum sensing alter the transcriptome and phenotype of Pseudomonas aeruginosa, and the composition of bacterial biofilms from wastewater treatment plants

Biofilms represent an essential way of life and colonization of new environments for microorganisms. This feature is regulated by quorum sensing (QS), a microbial communication system based on autoinducer molecules, such as N-acyl-homoserine lactones (AHLs) in Gram negative bacteria. In artificial ecosystems, like Wastewater Treatment Plants (WWTPs), biofilm attachment in filtration membranes produces biofouling. In this environment, the microbial communities are mostly composed of Gram-negative phyla. Thus, we used two AHLs-degrading enzymes, obtained from Actinoplanes utahensis (namely AuAAC and AuAHLA) to determine the effects of degradation of QS signals in the biofilm formation, among other virulence factors, of a Pseudomonas aeruginosa strain isolated from a WWTP, assessing molecular mechanisms through transcriptomics. Besides, we studied the possible effects on community composition in biofilms from activated sludge samples. Although the studied enzymes only degraded the AHLs involved in one of the four QS systems of P. aeruginosa, these activities produced the deregulation of the complete QS network. In fact, AuAAC -the enzyme with higher catalytic efficiency- deregulated all the four QS systems. However, both enzymes reduced the biofilm formation and pyocyanin and protease production. The transcriptomic response of P. aeruginosa affected QS related genes, moreover, transcriptomic response to AuAAC affected mainly to QS related genes. Regarding community composition of biofilms, as expected, the abundance of Gram-negative phyla was significantly decreased after enzymatic treatment. These results support the potential use of such AHLs-degrading enzymes as a method to reduce biofilm formation in WWTP membranes and ameliorate bacterial virulence.

Bacterial Composition of Biofilms of a Local Tigernut Drink Processing Unit in Yenagoa, Nigeria

Microorganisms grow and form biofilms on surfaces of equipment employed in food processing. These biofilms are considered as the major sources of contamination in the food industry. The study determined the bacterial load and composition on surfaces of equipment and utensils employed in tigernut drink production, from a vendor with a production unit located in an open market in Yenagoa, Nigeria. Swabs were taken from different sampling points made of varying materials—the grinding machine (metal), the collection bowls, water storage containers (thick plastic) and retail bottles (light plastic). These were analysed for total heterotrophic bacterial counts (THBC) and bacterial diversity using standard procedures. The THBC of water used in the drink production was also determined. The THBC of the sampling points ranged from log 3.28±0.06 cfu/cm2 to log 5.18±0.05 cfu/cm2, and log 5.13±0.07 cfu/ml for the water sample. The grinder and water recorded higher bacterial load with a statistically significant difference (P<0.05). The retail bottles had the least bacterial count. Shigella spp. were the most isolated (27.451%), then Salmonella spp. (21.568%) and E. coli (15.686%). Streptococcus spp., Staphylococcus spp. and Vibrio spp. were the least isolated (11.765% each). Shigella and Salmonella species occurred at all sampling points but not on the retail bottles. E. coli was present in the water collection bowls and retail bottles while Streptococcus, Staphylococcus and Vibrio were found only in the retail bottles. On diversity, four bacterial genera (39.216%) were isolated from the retail bottles, three (25.490%) from the water storage containers and two (17.647%) from the grinder and collection bowls. The microorganisms possibly had preference for attachment to the surfaces based on the material makeup and nutrient availability. Their occurrence and high numbers reflect the low level of hygiene employed prior to drink production. This could be of public health concern.

Diversity and Functions of Epilithic Riverine Biofilms

This article relates epilithic dry- and wet-seasonal bacterial biofilm composition to water quality along Río de la Sabana near Acapulco, Mexico. Samples were taken from various locations including nearly pristine upland locations, adjacent to residential floodplain developments, and immediately upstream from an estuarine lagoon. Bacterial composition was identified through sequential DNA analysis at the phylum, class, order, and family levels, with most of these categorized as heterotrophs, autotrophs, denitrifiers, nitrogen fixers, pathogens, and/or potential bioremediators based on generalized literature-sourced assignments. The results were interpreted in terms of location by extent of effluent pollution, and by dry versus wet seasonal changes pertaining to biofilm composition, related bacterial functions, and the following water quality parameters: temperature, pH, dissolved oxygen, biological and chemical oxygen demand, fecal and total bacteria counts, methylene blue active substances, electrical conductivity, and nitrite, nitrate, ammonium, sulfate, and phosphate concentrations. It was found that epilithic bacterial biofilm diversity was richest during the wet season, was more varied in abundance along the upland locations, and was dominated by Proteobacteria and Bacteroidetes with bioremediation and pathogen functions along effluent-receiving river locations. Low-abundance families associated with anaerobic and denitrifying functions were more prevalent during the wet season, while low-abundance families associated with aerobic, N2-fixing and pH-elevating functions were more prevalent during the dry season.

Chlorine-based DUWL disinfectant leads to a different microbial composition of water derived biofilms compared to H2O2-based chemical disinfectants in vitro.

BackgroundBiofilm formation in dental unit waterlines (DUWL) may lead to health risks for dental staff and patients. Therefore, dental unit waterlines need to be disinfected, for instance by using chemical disinfectants. However, the application of chemical disinfectants may lead to the selection of specific microorganisms. Therefore, the aim of our study was to assess the microbial composition of water-derived biofilms, after a continuous exposure to maintenance doses of commercially available chemical disinfectants, in vitro.MethodsThe AAA-model was used to grow water derived biofilms. The biofilms were subjected to the maintenance dose of each disinfectant. To determine the microbial composition, the V4 hypervariable region of the 16S rRNA gene was sequenced. The sequences were clustered in operational taxonomic units (OTUs).ResultsThe bacterial composition of biofilms in all treatment groups differed significantly (PERMANOVA F = 4.441, p = 0.001). Pairwise comparisons revealed Anoxyl treated biofilms were significantly different from all groups (p = 0.0001). In the Anoxyl-treated biofilms, the relative abundance of Comamonadaceae and Sphingopyxis was high compared to the Dentosept, Green and Clean and Oxygenal groups.ConclusionWe concluded that exposure to low doses of the chlorine-based chemical disinfectant Anoxyl led to a substantially different composition of water derived biofilms compared to biofilms exposed to H2O2-based chemical disinfectants.

Impact of algal organic matter on the performance, cyanotoxin removal, and biofilms of biologically-active filtration systems

The occurrence of harmful algal blooms dominated by toxic cyanobacteria has induced continuous loadings of algal organic matter (AOM) and toxins in drinking water treatment plants. However, the impact of AOM on the active biofilms and microbial community structures of biologically-active filtration (BAF), which directly affects the contaminant removal, is not well understood. In this study, we systematically examined the effects of AOM on BAF performance and bacterial biofilm formation over 240 days, tracing the removal of specific AOM components, a cyanotoxin [microcystin-LR (MC-LR)], and microbial community responses. The component analysis (excitation and emission matrix analysis) results for AOM revealed that terrestrial humic-like substances showed the highest removal among all the identified components and were strongly correlated to MC-LR removal. In addition, reduced empty bed contact time and deactivation of biofilms significantly decreased BAF performances for both AOM and MC-LR. The active biofilm, bacterial community structure, and mlrA gene (involved in microcystin degradation) abundance demonstrated that bacterial biofilm composition responded to AOM and MC-LR, in which Rhodocyclaceae, Saprospiraceae, and Comamonadaceae were dominant. In addition, MC-LR biodegradation appeared to be more active at the top than at the bottom layer in BAF. Overall, this study provides deeper insights into the role of biofilms and filter operation on the fate of AOM and MC-LR in BAF.

Impact of substrate material and chlorine/chloramine on the composition and function of a young biofilm microbial community as revealed by high-throughput 16S rRNA sequencing.

The bacterial composition of biofilms in drinking water distribution systems is significantly impacted by the disinfection regime and substrate material. However, studies that have addressed the changes in the biofilm community during the early stage of formation (less than 10 weeks) were not yet adequate. Here, we explore the effects of the substrate materials (cast iron, stainless steel, copper, polyvinyl chloride, and high density polyethylene) and different disinfectants (chlorine and chloramine) on the community composition and function of young biofilm by using 16S rDNA sequencing. The results showed that Alphaproteobacteria (39.14%-80.87%) and Actinobacteria (5.90%-40.03%) were the dominant classes in chlorine-disinfection samples, while Alphaproteobacteria (17.46%-74.18%) and Betaproteobacteria (3.79%-68.50%) became dominant in a chloraminated group. The infrequently discussed genus Phreatobacter became predominant in the chlorinated samples, but it was inhibited by chloramine and copper ions. The key driver of the community composition was indicated as different disinfectants according to principle coordination analysis (PCoA) and Permutational multivariate analysis of variance (Adonis test), and the bacterial community changed significantly over time. Communities of biofilms grown on cast iron showed a great distance from the other materials according to Bray-Curtis dissimilarity, and they had a unique dominant genus, Dechloromonas. A metagenomics prediction based on 16S rDNA was used to detect the functional pathways of antibiotic biosynthesis and beta-lactam resistance, and it revealed that several pathways were significantly different in terms of their chlorinated and chloraminated groups.

Bacterial composition of biofilms formed on dairy-processing equipment

Polymicrobial biofilms often form on the surfaces of food-processing machinery, causing equipment damage and posing a contamination risk for the foods processed by the system. The composition of the microbial communities that make up these biofilms is largely unknown, especially in the dairy industry. To address this deficit, we investigated the bacterial composition of biofilms that form on the surfaces of equipment during dairy processing using Illumina MiSeq sequencing and culture-dependent methods. Illumina sequencing identified eight phyla, comprising six classes, ten orders, fifteen families, eighteen genera, and eighteen species. In contrast, only eight species were isolated from the same samples using the culture-based method. To determine the ability of the identified bacteria to form biofilms, biofilm formation analysis via crystal violet staining was performed. Five of the eight culturable species, Acinetobacter baumannii, Acinetobacter junii, Enterococcus faecalis, Corynebacterium callunae, and Stenotrophomonas maltophilia, were able to form biofilms. Since most of the identified bacteria are potential food-borne or opportunistic pathogens, this study provides guidance for quality control of products produced in dairy processing facilities.

Deep neural networks outperform human expert's capacity in characterizing bioleaching bacterial biofilm composition

BackgroundDeep neural networks have been successfully applied to diverse fields of computer vision. However, they only outperform human capacities in a few cases. MethodsThe ability of deep neural networks versus human experts to classify microscopy images was tested on biofilm colonization patterns formed on sulfide minerals composed of up to three different bioleaching bacterial species attached to chalcopyrite sample particles. ResultsA low number of microscopy images per category (<600) was sufficient for highly efficient computational analysis of the biofilm's bacterial composition. The use of deep neural networks reached an accuracy of classification of ∼90% compared to ∼50% for human experts. ConclusionsDeep neural networks outperform human experts’ capacity in characterizing bacterial biofilm composition involved in the degradation of chalcopyrite. This approach provides an alternative to standard, time-consuming biochemical methods.

Bacterial biofilm composition in healthy subjects with and without caries experience.

ABSTRACTObjective:The composition of the oral microbiome differs distinctively between subjects with and without active caries. Still, caries research has mainly been focused on states of disease; aspects about how biofilm composition and structure maintain oral health still remain widely unclear. Therefore, the aim of the study was to compare the healthy oral microbiome of caries-free adult subjects with and without former caries experience using next generation sequencing methods.Methods: 46 samples were collected from subjects without any signs of untreated active caries. Samples of pooled supragingival plaque from 19 subjects without caries experience (NH; DMFT = 0) and 27 subjects with ‘caries experience’ (CE; DMFT > 0 [F(T)> 0; D(T)= 0]) were analyzed by 16S ribosomal RNA amplicon sequencing.Results: Subjects with caries experience did not exhibit a dramatically modified supragingival plaque microbiome. However, we observed a slight and significant modification between the two groups, validated by PERMANOVA (NH vs. CE: R2 0.04; p= 0.039). The composition of the microbiome of subjects with caries experience indicates a tendency to lower α-diversity and richness. Subjects without caries experience showed a significant higher evenness compared to patients with previous caries. LDA effect size (LEfSe) analysis demonstrated that the genus Haemophilus is significantly more frequent in patients with caries experience. For the group without caries experience LefSe analysis showed a set of 11 genera being significantly more frequent, including Corynebacterium, Fusobacterium, Capnocytophaga, Porphyromonas, Prevotella,and Leptotrichia.Conclusion: The analysis of the oral microbiome of subjects with and without caries experience indicates specific differences. With the presence of Corynebacterium and Fusobacterium subjects without caries experience exhibited more frequently organisms that are considered to be main actors in structural plaque formation and integration. The abundance of Corynebacterium might be interpreted as a signature for dental health.

Microbiome Analysis of Biofilms of Silver Nanoparticle-Dispersed Silane-Based Coated Carbon Steel Using a Next-Generation Sequencing Technique.

Previously, we demonstrated that silver nanoparticle-dispersed silane-based coating could inhibit biofilm formation in conditions where seawater was used as a bacterial source and circulated in a closed laboratory biofilm reactor. However, it is still unclear whether the microbiome of a biofilm of silver nanoparticle-dispersed silane-based coating samples (Ag) differs from that of a biofilm of non-dispersed silane-based coating samples (Non-Ag). This study aimed to perform a microbiome analysis of the biofilms grown on the aforementioned coatings using a next-generation sequencing (NGS) technique. For this, a biofilm formation test was conducted by allowing seawater to flow through a closed laboratory biofilm reactor; subsequently, DNAs extracted from the biofilms of Ag and Non-Ag were used to prepare 16S rRNA amplicon libraries to analyze the microbiomes by NGS. Results of the operational taxonomy unit indicated that the biofilms of Non-Ag and Ag comprised one and no phyla of archaea, respectively, whereas Proteobacteria was the dominant phylum for both biofilms. Additionally, in both biofilms, Non-Ag and Ag, Marinomonas was the primary bacterial group involved in early stage biofilm formation, whereas Anaerospora was primarily involved in late-stage biofilm formation. These results indicate that silver nanoparticles will be unrelated to the bacterial composition of biofilms on the surface of silane-based coatings, while they control biofilm formation there.

Bacterial Colonization and Tissue Compatibility of Denture Base Resins.

Currently, there is minimal clinical data regarding biofilm composition on the surface of denture bases and the clinical tissue compatibility. Therefore, the aim of this experimental study was to compare the bacterial colonization and the tissue compatibility of a hypoallergenic polyamide with a frequently used PMMA resin tested intraorally in a randomized split-mouth design. Test specimens made of polyamide (n = 10) and PMMA (n = 10) were attached over a molar band appliance in oral cavity of 10 subjects. A cytological smear test was done from palatal mucosa at baseline and after four weeks. The monolayers were inspected for micronuclei. After four weeks in situ, the appliance was removed. The test specimens were immediately cultivated on non-selective and selective nutrient media. All growing colonies were identified using VITEK-MS. The anonymized results were analyzed descriptively. A total of 110 different bacterial species could be isolated, including putative pathogens. An average of 17.8 different bacterial species grew on the PMMA specimens, and 17.3 on the polyamide specimens. The highest number of different bacterial species was n = 24, found on a PMMA specimen. On the two specimens, a similar bacterial distribution was observed. Micronuclei, as a marker for genotoxic potential of dental materials, were not detected. This study indicates that the composition of bacterial biofilm developed on these resins after four weeks is not influenced by the type of resin itself. The two materials showed no cytological differences. This investigation suggests that polyamide and PMMA are suitable for clinical use as denture base material.

Effects of low dose silver nanoparticle treatment on the structure and community composition of bacterial freshwater biofilms.

The application of engineered silver nanoparticles (AgNPs) in a considerable amount of registered commercial products inevitably will result in the continuous release of AgNPs into the natural aquatic environment. Therefore, native biofilms, as the prominent life form of microorganisms in almost all known ecosystems, will be subjected to AgNP exposure. Despite the exponentially growing research activities worldwide, it is still difficult to assess nanoparticle-mediated toxicity in natural environments. In order to obtain an ecotoxicologically relevant exposure scenario, we performed experiments with artificial stream mesocosm systems approaching low dose AgNP concentrations close to predicted environmental concentrations. Pregrown freshwater biofilms were exposed for 14 days to citrate-stabilized AgNPs at a concentration of 600 μg l-1 in two commonly used sizes (30 and 70 nm). Sublethal effects of AgNP treatment were assessed with regard to biofilm structure by gravimetric measurements (biofilm thickness and density) and by two biomass parameters, chlorophyll a and protein content. The composition of bacterial biofilm communities was characterized by t-RFLP fingerprinting combined with phylogenetic studies based on the 16S gene. After 14 days of treatment, the structural parameters of the biofilm such as thickness, density, and chlorophyll a and protein content were not statistically significantly changed by AgNP exposure. Furthermore, t-RFLP fingerprint analysis showed that the bacterial diversity was not diminished by AgNPs, as calculated by Shannon Wiener and evenness indices. Nevertheless, t-RFLP analysis also indicated that AgNPs led to an altered biofilm community composition as was shown by cluster analysis and multidimensional scaling (MDS) based on the Bray Curtis index. Sequence analysis of cloned 16S rRNA genes further revealed that changes in community composition were related with the displacement of putatively AgNP-sensitive bacterial taxa Actinobacteria, Chloroflexi, and Cyanobacteria by taxa known for their enhanced adaptability towards metal stress, such as Acidobacteria, Sphingomonadales, and Comamonadaceae. This measurable community shift, even after low dose AgNP treatment, causes serious concerns with respect to the broad application of AgNPs and their potentially adverse impact on the ecological function of lotic biofilms, such as biodegradation or biostabilization.

Effect of Probiotic Lactobacilli on the Growth of Streptococcus Mutans and Multispecies Biofilms Isolated from Children with Active Caries.

BackgroundThe purpose of this investigation was to evaluate the effect of probiotic lactobacilli on Streptococcus mutans (MS) and multispecies biofilms isolated from children with severe caries.Material/MethodsTwenty children with active caries (DMFS ≥6) were selected as the experimental group and Streptococcus mutans (MS) were isolated from their saliva. After identification the MS strains were mixed with lactobacilli at 37°C, following which viable MS colonies were counted. At the same time dental plaques from the children were mixed with lactobacilli in vitro to form biofilms, and the population of nine common strains in the biofilms was enumerated after 24 hours of growth.ResultsLactobacillus casei Shirota, L. casei LC01, L. plantarum ST-III and L. paracasei LPC37 all had strong inhibitory effects on the majority the MS isolated from children with active caries, with the inhibition rate reaching approximately 70–90% (p<0.05). L. casei Shirota, L. casei LC01, L. plantarum ST-III, L. paracasei LPC37 also significantly reduced the numbers of MS, Streptococcus spp., S. sanguinis and total bacteria in mixed biofilms compared with the control group (p<0.05).ConclusionsThe four strains of lactobacilli were able to inhibit the growth of Streptococcus mutans and had effects on the composition of bacterial biofilms in vitro. Ingestion of probiotics may be a promising method of caries prevention.