Biofilm Response Research Articles

High fluoride resistance and virulence profile of environmental Pseudomonas isolated from water sources.

In our previous study, all Pseudomonas strains THP6, THP41, and OHP5 were identified as fluoride-resistant bacteria isolated from Dindigul district, Tamilnadu, India. The selected strains exhibiting a high level of fluoride resistance was determined in Luria broth (LB) medium and LB agar plates. In a further effort, fluoride-resistant organisms were tested for hemolytic activity and showed β-hemolysis on blood agar plates. The virulence factors such as gyrB, toxA, algD and lasB, plcH, rhlC and biofilm response genes (pslA, pelA, ppyR) were detected by PCR analysis. The putative genus-specific and species-specific PCR also confirmed that the selected fluoride-resistant strains were belonging to Pseudomonas aeruginosa species. Fluoride-resistance gene crcB was amplified by gene-specific primers. The crcB gene was cloned in TA vector and transformed into E. coli DH5α. Comparative and blast analysis of THP6, THP41, and OHP5 strains crcB gene sequences were high homology with P. aeruginosa fluoride efflux transporter crcB and P. aeruginosa putative fluoride ion transporter crcB. The recombinants were efficiently growing in the NaF containing LB agar plates. The fluoride tolerance of these strains was also associated with resistance to multiple antibiotics. These results can lead to the use of the fluoride resistance gene of P. aeruginosa for the development of a biosensor for fluoride detection.

Longitudinal optical coherence tomography to visualize the in vivo response of middle ear biofilms to antibiotic therapy

Studying the impact of antibiotic treatment on otitis media (OM), the leading cause of primary care office visits during childhood, is critical to develop appropriate treatment strategies. Tracking dynamic middle ear conditions during antibiotic treatment is not readily applicable in patients, due to the limited diagnostic techniques available to detect the smaller amount and variation of middle ear effusion (MEE) and middle ear bacterial biofilm, responsible for chronic and recurrent OM. To overcome these challenges, a handheld optical coherence tomography (OCT) system has been developed to monitor in vivo response of biofilms and MEEs in the OM-induced chinchilla model, the standard model for human OM. As a result, the formation of MEE as well as biofilm adherent to the tympanic membrane (TM) was longitudinally assessed as OM developed. Various types of MEEs and biofilms in the chinchilla model were identified, which showed comparable features as those in humans. Furthermore, the effect of antibiotics on the biofilm as well as the amount and type of MEEs was investigated with low-dose and high-dose treatment (ceftriaxone). The capability of OCT to non-invasively track and examine middle ear conditions is highly beneficial for therapeutic OM studies and will lead to improved management of OM in patients.

Microbial Functional Responses in Marine Biofilms Exposed to Deepwater Horizon Spill Contaminants.

Marine biofilms are essential biological components that transform built structures into artificial reefs. Anthropogenic contaminants released into the marine environment, such as crude oil and chemical dispersant from an oil spill, may disrupt the diversity and function of these foundational biofilms. To investigate the response of marine biofilm microbiomes from distinct environments to contaminants and to address microbial functional response, biofilm metagenomes were analyzed from two short-term microcosms, one using surface seawater (SSW) and the other using deep seawater (DSW). Following exposure to crude oil, chemical dispersant, and dispersed oil, taxonomically distinct communities were observed between microcosms from different source water challenged with the same contaminants and higher Shannon diversity was observed in SSW metagenomes. Marinobacter, Colwellia, Marinomonas, and Pseudoalteromonas phylotypes contributed to driving community differences between SSW and DSW. SSW metagenomes were dominated by Rhodobacteraceae, known biofilm-formers, and DSW metagenomes had the highest abundance of Marinobacter, associated with hydrocarbon degradation and biofilm formation. Association of source water metadata with treatment groups revealed that control biofilms (no contaminant) harbor the highest percentage of significant KEGG orthologs (KOs). While 70% functional similarity was observed among all metagenomes from both experiments, functional differences between SSW and DSW metagenomes were driven primarily by membrane transport KOs, while functional similarities were attributed to translation and signaling and cellular process KOs. Oil and dispersant metagenomes were 90% similar to each other in their respective experiments, which provides evidence of functional redundancy in these microbiomes. When interrogating microbial functional redundancy, it is crucial to consider how composition and function evolve in tandem when assessing functional responses to changing environmental conditions within marine biofilms. This study may have implications for future oil spill mitigation strategies at the surface and at depth and also provides information about the microbiome functional responses of biofilms on steel structures in the marine built environment.

Proteomic Analysis of Caspofungin-Induced Responses in Planktonic Cells and Biofilms of Candida albicans.

Candida albicans biofilms display markedly increased antifungal resistance, and the underlying mechanisms remain unclear. This study investigated the signature profiles of C. albicans planktonic cells and biofilms in response to caspofungin (CAS) by mass spectrometry-based shotgun proteomics. We found that C. albicans biofilms were twofold more resistant to CAS with reference to planktonic cells. Notably, 9.6% of C. albicans biofilm cells survived the lethal treatment of CAS (128 μg/ml), confirmed by LIVE/DEAD staining, confocal laser scanning microscopy (CLSM) and scanning electron microscopy analyses. The responses of C. albicans planktonic cells and biofilms to CAS treatment at respective minimum inhibitory concentrations (MICs) were assessed by high-throughput proteomics and bioinformatics approaches. There were 148 and 224 proteins with >twofold difference identified from the planktonic cells and biofilms, respectively. CAS treatment downregulated several cell wall- and oxidative stress-related proteins. Whereas, CAS-induced action was compensated by markedly increased expression of many other proteins involved in cell wall integrity and stress response (e.g., heat shock proteins). Moreover, considerable expression changes were identified in metabolism-associated proteins like glycolysis, tricarboxylic acid (TCA) cycle and ATP biosynthesis. Importantly, various key proteins for cell wall integrity, stress response and metabolic regulation (e.g., PIL1, LSP1, HSP90, ICL1, and MLS1) were exclusively enriched and implicated in C. albicans biofilms. This study demonstrates that C. albicans biofilms undergo highly complicated yet complex regulation of multiple cellular pathways in response to CAS. Signature proteins essential for modulating cell wall integrity, stress response and metabolic activities may account for the antifungal resistance of C. albicans biofilms.

Ecotoxicological effects of erythromycin on a multispecies biofilm model, revealed by metagenomic and metabolomic approaches.

The presence of antibiotics such as erythromycin, even in trace amounts, has long been acknowledged for negatively impacting ecosystems in freshwater environments. Although many studies have focused on the impact of antibiotic pollution at a macroecological level, the impact of erythromycin on microecosystems, such as freshwater biofilms, is still not fully understood. This knowledge gap may be attributed to the lack of robust multispecies biofilm models for fundamental investigations. Here, we used a lab-cultured multispecies biofilm model to elucidate the holistic response of a microbial community to erythromycin exposure using metagenomic and metabolomic approaches. Metagenomic analyses revealed that biofilm microbial diversity did not alter following erythromycin exposure. Notably, certain predicted metabolic pathways such as cell-cell communication pathways, amino acid metabolism, and peptidoglycan biosynthesis, mainly by the phyla Actinobacteria, Alpha/Beta-proteobacteria, Bacteroidetes, and Verrucomicrobia, were found to be involved in the maintenance of homeostasis-like balance in the freshwater biofilm. Further untargeted metabolomics data highlighted changes in lipid metabolism and linoleic acid metabolism and their related molecules as a direct consequence of erythromycin exposure. Overall, the study presented a unique picture of how multispecies biofilms respond to single environmental stress exposures. Moreover, the study demonstrated the feasibility of using lab simulated multispecies biofilms for investigating their interaction and reactivity of specific bioactive compounds or pollutants at a fundamental level.

Microbial Metabolic Genes Crucial for S. aureus Biofilms: An Insight From Re-analysis of Publicly Available Microarray Datasets.

Bacterial biofilms are microbial lifestyles found in all environments. Up to 80% of human infections and 60–70% of hospital-acquired infections have a biofilm origin, with Staphylococcus aureus one of the leading causes of these infections. Microorganisms in biofilms exhibit significant antimicrobial resistance which poses important treatment challenges, hence the urgent need to identify novel antibiofilm strategies. Microbes form biofilms in response to various factors, and once these 3-dimentional structures form they are highly recalcitrant to removal. The switch from planktonic lifestyle to the biofilm protected mode of growth results in a phenotypic shift in the behavior of the microorganisms in terms of growth rate and gene expression. Given these changes, investigation of microbial gene expression and their modulation at different stages of biofilm maturation is needed to provide vital insight into the behavior of biofilm cells. In this study, we analyzed publicly available transcriptomic dataset of S. aureus biofilms at different stages of maturation to identify consistently upregulated genes irrespective of the biofilm maturation stage. Our reanalysis identified a total of 6 differentially expressed genes upregulated in both 48 and 144-h old S. aureus biofilms. Functional analysis revealed that these genes encode for proteins which play a role in key microbial metabolic pathways. However, these genes, as yet, are unrelated or fully studied in the context of biofilm. Moreover, the findings of this in silico work, suggest that these genes may represent potential novel targets for the development of more effective antibiofilm strategies against S. aureus biofilm-associated infections.

Reduction of alternative electron acceptors drives biofilm formation in Shewanella algae

Shewanella spp. possess a broad respiratory versatility, which contributes to the occupation of hypoxic and anoxic environmental or host-associated niches. Here, we observe a strain-specific induction of biofilm formation in response to supplementation with the anaerobic electron acceptors dimethyl sulfoxide (DMSO) and nitrate in a panel of Shewanella algae isolates. The respiration-driven biofilm response is not observed in DMSO and nitrate reductase deletion mutants of the type strain S. algae CECT 5071, and can be restored upon complementation with the corresponding reductase operon(s) but not by an operon containing a catalytically inactive nitrate reductase. The distinct transcriptional changes, proportional to the effect of these compounds on biofilm formation, include cyclic di-GMP (c-di-GMP) turnover genes. In support, ectopic expression of the c-di-GMP phosphodiesterase YhjH of Salmonella Typhimurium but not its catalytically inactive variant decreased biofilm formation. The respiration-dependent biofilm response of S. algae may permit differential colonization of environmental or host niches.

Temperature modulates stress response in mainstream anammox reactors

Autotrophic nitrogen removal by anaerobic ammonium oxidizing (anammox) bacteria is an energy-efficient nitrogen removal process in wastewater treatment. However, full-scale deployment under mainstream conditions remains challenging for practitioners due to the high stress susceptibility of anammox bacteria towards fluctuations in dissolved oxygen (DO) and temperature. Here, we investigated the response of microbial biofilms with verified anammox activity to DO shocks under 20 °C and 14 °C. While pulse disturbances of 0.3 mg L−1 DO prompted only moderate declines in the NH4+ removal rates, 1.0 mg L−1 DO led to complete but reversible inhibition of the NH4+ removal activity in all reactors. Genome-centric metagenomics and metatranscriptomics were used to investigate the stress response on various biological levels. We show that temperature regime and strength of DO perturbations induced divergent responses from the process level down to the transcriptional profile of individual taxa. Community-wide gene expression differed significantly depending on the temperature regime in all reactors, and we found a noticeable impact of DO disturbances on genes involved in transcription, translation, replication and posttranslational modification at 20 °C but not 14 °C. Genome-centric analysis revealed that different anammox species and other key biofilm taxa differed in their transcriptional responses to distinct temperature regimes and DO disturbances.

Numerical modeling of the mechanical response of bacterial biofilm to flow by using an SPH poroviscoelastic model

AbstractWe present a weakly‐compressible two‐phase poroviscoelastic biofilm model based on Smoothed Particle Hydrodynamics (SPH) method. The biofilm is modeled as a mixture of solid and fluid phases interacting via drag forces. Benefiting from the Lagrangian feature of the SPH method, large deformations of the solid structure can be modeled straightforwardly.

Influence of caries activity and number of saliva donors: mineral and microbiological responses in a microcosm biofilm model.

Objective this study evaluated the mineral and microbiological response of biofilms originating from different types of saliva inoculum with distinct levels of caries activity.Methodology the biofilms grown over enamel specimens originated from saliva collected from a single donor or five donors with two distinct levels of caries activity (caries-active and caries-free) or from pooling saliva from ten donors (five caries-active and five caries-free). The percentage surface hardness change (%SHC) and microbiological counts served as outcome variables.Results the caries activity of donors did not affect the %SHC values. Inoculum from five donors compared to a single donor showed higher %SHC values (p=0.019). Higher lactobacilli counts were observed when saliva from caries-active donors was used as the inoculum (p=0.017). Pooled saliva from both caries activity levels showed higher mutans streptococci counts (p<0.017).Conclusion Overall, pooled saliva increased the mineral response of the derived biofilms, but all the inoculum conditions formed cariogenic biofilms and caries lesions independently of caries activity.

Measurement of free chlorine levels in water using potentiometric responses of biofilms and applications for monitoring and managing the quality of potable water

Residual free chlorine is not monitored continuously at scale in drinking water distribution systems because existing real-time sensor technologies require frequent maintenance, cleaning, and calibration, which makes these products too costly to be used throughout a distribution system. As a result, current measurement approaches require manual sampling, which is not feasible for the consistent monitoring of free chlorine because chlorine concentrations vary significantly throughout pipeline distribution and over time and space. This research presents an alternative and cost-effective method of predicting free chlorine levels in drinking water using graphite electrodes coated with naturally grown microbial biofilms. This Microbial Potentiometric Sensor (MPS) array was installed in a Continuously Mixed Batch Reactor (CMBR), and drinking water containing variable free chlorine concentrations. The chlorine concentrations were introduced in a controlled manner, and the MPS signals were monitored over time. MPS signals were measured from the change in Open Circuit Potential (OCP) across the MPS array in real-time. An empirically derived relationship between the normalized change in OCP and free chlorine was established by fitting individual and average MPS data to a decaying exponential growth function in order to predict free chlorine levels. The results show that free chlorine can be predicted with reasonable accuracy, with model validation showing an average absolute error of ±0.09 ppm below 1.1 ppm and ±0.30 ppm between 1.1 and 2.7 ppm. However, the accuracy of predictions was reduced at higher free chlorine levels. The researchers conclude that MPS systems may benefit drinking water distribution systems by measuring free chlorine. These advantages of the MPS are especially pronounced in the developing world because this system is inexpensive and does not require routine maintenance or cleaning. The system relies on a naturally forming and regenerating biofilm and an inexpensive potentiometric meter to produce stable measurements.

Characterization of river biofilm responses to the exposure with heavy metals using a novel micro fluorometer biosensor

River biofilms are a suitable indicator of toxic stress in aquatic ecosystems commonly exposed to various anthropogenic pollutants from industrial, domestic, and agricultural sources. Among these pollutants, heavy metals are of particular concern as they are known to interfere with various physiological processes of river biofilm, directly or indirectly related to photosynthetic performance. Nevertheless, only limited toxicological data are available on the mechanisms and toxicodynamics of heavy metals in biofilms. Pulse Amplitude Modulated (PAM) fluorometry is a rapid, non-disruptive, well-established technique to monitor toxic responses on photosynthetic performance, fluorescence-kinetics, and changes in yield in other non-photochemical processes. In this study, a new micro-PAM-sensor was tested to assess potential acute and chronic effects of heavy metals in river biofilm. Toxicity values across the three parameters considered in this study (photosynthetic yield YII, non-photochemical quenching NPQ, and basal fluorescence F0) were comparable, as determined EC50 were within one order of magnitude (EC50 ∼1−10 mg L−1). However, the stimulation of NPQ was more clearly associated with early acute effects, especially in illuminated samples, while depression of YII and F0 were more prevalent in chronic tests. These results have implications for the development of functional indicators for the biomonitoring of aquatic health, in particular for the use of river biofilm as a bioindicator of water quality. In conclusion, the approach proposed seems promising to characterize and monitor the exposure and impact of heavy metals on river periphyton communities. Furthermore, this study provides a fast, highly sensitive, inexpensive, and accurate laboratory method to test effects of pollutants on complex periphyton communities that can also give insights regarding the probable toxicological mechanisms of heavy metals on photosynthetic performance in the river biofilm.

Staphylococcus aureus Biofilms and Their Response to a Relevant in vivo Iron Source.

Biofilm infections can be chronic, life threatening and challenging to eradicate. Understanding in vivo stimuli affecting the biofilm cycle is one step toward targeted prevention strategies. Iron restriction by the host is a stimulus for biofilm formation for some Staphylococcus aureus isolates; however, in some infection scenarios bacteria are exposed to abundant amounts of hemoglobin (Hb), which S. aureus is able to use as iron source. Thus, we hypothesized a role for Hb in the biofilm infection. Microplate “biofilm” assays showed biofilm-matrix production was increased in the presence of hemoglobin when compared to the provision of iron as an inorganic salt. Microscopic analysis of biofilms showed that the provision of iron as hemoglobin consistently caused thicker and more structured biofilms when compared to the effect of the inorganic iron source. Iron responsive biofilm gene expression analysis showed that Agr Quorum Sensing, a known biofilm dispersal marker, was repressed with hemoglobin but induced with an equivalent amount of inorganic iron in the laboratory strain Newman. The gene expression of two biofilm structuring agents, PSMα and PSMβ, differed in the response to the iron source provided and was not correlated to hemoglobin-structured biofilms. A comparison of the model pathogen S. aureus Newman with local clinical isolates demonstrated that while there was a similar phenotypic biofilm response to hemoglobin, there was substantial variation in the expression of key biofilm dispersal markers, suggesting an underappreciated variation in biofilm regulome among S. aureus isolates and that no general inferences can be made by studying the behavior of single strains.

Antibacterial response of oral microcosm biofilm to nano-zinc oxide in adhesive resin

ObjectiveVarious nanoparticles are currently under investigation to impart biointeractivity for dental materials. This study aimed to: (1) formulate an experimental dental adhesive containing ZnO nanoparticles; (2) evaluate its chemical and mechanical properties; and (3) assess the antibacterial response against oral microcosm biofilm. MethodsNanosized ZnO was chemically and morphologically evaluated. ZnO was incorporated at 0 (GCTRL), 2.5 (G2.5%), 5 (G5%) and 7.5 (G5%) wt.% in an experimental dental adhesive. The adhesives were evaluated for the degree of conversion (DC), flexural strength (FS), and elastic modulus (E). The antibacterial activity was evaluated using a 48h-microcosm biofilm model after the formation of acquired pellicle on samples’ surfaces. Colony-forming units (CFU), metabolic activity, and live/dead staining were assessed. ResultsNanosized ZnO presented characteristic peaks of Zn-O bonds, and the particles were arranged in agglomerates. The DC ranged from 62.21 (±1.05) % for GCtrl to 46.15 (±1.23) % for G7.5% (p<0.05). G7.5% showed lower FS compared to all groups (p<0.05). Despite achieving higher E (p<0.05), G2.5% did not show differences for GCtrl regarding the FS (p>0.05). G7.5% had lower CFU/mL compared to GCtrl for mutans streptococci (p<0.05) and total microorganisms (p<0.05), besides presenting lower metabolic activity (p<0.05) and higher dead bacteria via biofilm staining. SignificanceThe dental adhesives' physicochemical properties were similar to commercial adhesives and in compliance with ISO recommendations. G7.5% restricted the growth of oral microcosm biofilm without impairing the physicochemical performance.

Pseudomonas putida actively forms biofilms to protect the population under antibiotic stress.

Antibiotics are frequently used for clinical treatment and by the farming industry, and most of these are eventually released into the surrounding environment. The impact of these antibiotic pollutants on environmental microorganisms is a concern. The present study showed that after Pseudomonas putida entered the logarithmic growth phase, tetracycline strongly stimulated its biofilm formation in a dose-dependent manner. This was supported by the increased expression of the key adhesin gene lapA in response to tetracycline treatment. Tetracycline treatment also changed the expression levels of the exopolysaccharide gene clusters alg, bcs and pea and the adhesin gene lapF. However, these genes did not participate in the tetracycline-induced biofilm formation. When a biofilm had been established, the P.putida population became more tolerant to tetracycline. Confocal laser scanning microscopic images showed that the interior of the biofilm provided favorable conditions that protected bacterial cells from tetracycline. Besides, biofilm formation of P.putida was also promoted by several other antibiotics, including oxytetracycline, fluoroquinolones, rifampicin, and imipenem, but not aminoglycosides. Susceptibility tests suggested that biofilm conferred a higher tolerance on P.putida cells to specific antibiotics (e.g., tetracyclines and fluoroquinolones). These antibiotics exerted a stronger inducing effect on biofilm formation. Together, our results indicate that P.putida actively forms robust biofilms in response to antibiotic stress, and the biofilms improve the survival of bacterial population under such stress.

Electrospinned bioactive glass/nano hydroxyapatite reinforced polycaprolactone GTR/GBR membranes in periodontics- A review

Periodontitis, a chronic inflammatory infectious disease triggered by an interaction between microbial biofilm and host response leading to gingival bleeding, connective tissue destruction , alveolar bone loss and finally causing tooth loss. Periodontal regeneration or restoration is the ultimate goal of any periodontal treatment. This can be achieved by GTR/GBR [Guided Tissue Regeneration/Guided Bone Regeneration] membranes along with bone replacement grafts. With the evolution of Tissue engineering and different generation of GTR/GBR nanofibrous membranes, periodontal restoration or regeneration can be easily and rapidly achieved . Nanofibrous GTR/GBR Membranes can be fabricated using natural or synthetic polymers by different techniques such as Phase separation, Self assembly and Electrospinning [E-spinning]. Of these, E-spinning is the most widely studied technique and also seems to exhibit the most promising results for Tissue Engineering[TE] applications . This review addresses the Electrospinning method of GTR/GBR membrane fabrication using Bioactive Glass [BG]/Nano Hydroxyapatite[nHAP] reinforced polycaprolactone polymer and its advantages and disadvantages. Keywords: Electrospinning, Guided Tissue Regeneration, Periodontal Regeneration

Modelling experimentally measured of ciprofloxacin antibiotic diffusion in Pseudomonas aeruginosa biofilm formed in artificial sputum medium.

We study the experimentally measured ciprofloxacin antibiotic diffusion through a gel-like artificial sputum medium (ASM) mimicking physiological conditions typical for a cystic fibrosis layer, in which regions occupied by Pseudomonas aeruginosa bacteria are present. To quantify the antibiotic diffusion dynamics we employ a phenomenological model using a subdiffusion-absorption equation with a fractional time derivative. This effective equation describes molecular diffusion in a medium structured akin Thompson’s plumpudding model; here the ‘pudding’ background represents the ASM and the ‘plums’ represent the bacterial biofilm. The pudding is a subdiffusion barrier for antibiotic molecules that can affect bacteria found in plums. For the experimental study we use an interferometric method to determine the time evolution of the amount of antibiotic that has diffused through the biofilm. The theoretical model shows that this function is qualitatively different depending on whether or not absorption of the antibiotic in the biofilm occurs. We show that the process can be divided into three successive stages: (1) only antibiotic subdiffusion with constant biofilm parameters, (2) subdiffusion and absorption of antibiotic molecules with variable biofilm transport parameters, (3) subdiffusion and absorption in the medium but the biofilm parameters are constant again. Stage 2 is interpreted as the appearance of an intensive defence build–up of bacteria against the action of the antibiotic, and in the stage 3 it is likely that the bacteria have been inactivated. Times at which stages change are determined from the experimentally obtained temporal evolution of the amount of antibiotic that has diffused through the ASM with bacteria. Our analysis shows good agreement between experimental and theoretical results and is consistent with the biologically expected biofilm response. We show that an experimental method to study the temporal evolution of the amount of a substance that has diffused through a biofilm is useful in studying the processes occurring in a biofilm. We also show that the complicated biological process of antibiotic diffusion in a biofilm can be described by a fractional subdiffusion-absorption equation with subdiffusion and absorption parameters that change over time.

Attachment and antibiotic response of early-stage biofilms studied using resonant hyperspectral imaging

Many bacterial species readily develop biofilms that act as a protective matrix against external challenge, e.g., from antimicrobial treatment. Therefore, biofilms are often responsible for persistent and recurring infections. Established methods for studying biofilms are either destructive or focus on the biofilm’s surface. A non-destructive method that is sensitive to the underside of the biofilm is highly desirable, as it allows studying the penetration of antibiotics through the film. Here, we demonstrate that the high surface sensitivity of resonant hyperspectral imaging provides this capability. The method allows us to monitor the early stages of Escherichia coli biofilm formation, cell attachment and microcolony formation, in-situ and in real-time. We study the response of the biofilm to a number of different antibiotics and verify our observations using confocal microscopy. Based on this ability to closely monitor the surface-bound cells, resonant hyperspectral imaging gives new insights into the antimicrobial resistance of biofilms.

Multicellular and unicellular responses of microbial biofilms to stress.

Biofilms are a ubiquitous mode of microbial life and display an increased tolerance to different stresses. Inside biofilms, cells may experience both externally applied stresses and internal stresses that emerge as a result of growth in spatially structured communities. In this review, we discuss the spatial scales of different stresses in the context of biofilms, and if cells in biofilms respond to these stresses as a collection of individual cells, or if there are multicellular properties associated with the response. Understanding the organizational level of stress responses in microbial communities can help to clarify multicellular functions of biofilms.

Modulation of Oral Biofilm and Immune Response Associated to Mucosa with Probiotic Bacteria as a Potential Approach in the Prevention of Dental Caries: A Systematic Review

Background: A variety of approaches have been developed for the control of dental caries, a pathology with high incidence and prevalence worldwide. The use of probiotic strains for the modulation of dental biofilm in the prevention of caries has been studied, but the available evidence shows varied methodologies; and the strains tested differ from one study to another. Objective: To analyse through a systematic review of clinical trials, the efficacy of using probiotic strains to prevent dental caries. Methods: A search was made in the scientific bases PubMed, Cochrane and Science Direct, prioritizing randomized double and triple-blind clinical trials from 2010 to 2020, including a total of 20 studies to be analysed. The selection criteria were consistent with the Preferred Reporting Items for Systematic Reviews protocol. Results: Within the studies, different types of probiotic bacteria were analysed, dominating Lactobacillus Paracasei. While most clinical trials show a favorable response in terms of a significant reduction of Streptococcus mutans in the oral microbiota, very few studies evaluated salivary pH and sIgA levels. Conclusion: The heterogeneity of the studies analysed and the multifactorial nature of dental caries do not allow us to ensure that probiotic therapy is completely effective in preventing this pathology. Although probiotic therapy can help by regulating the microbiological factor, there are other determinants that can favor the development of caries and that are barely approached in their relationship with bacteriotherapy. Future studies that homogeneously evaluate the use of probiotics could give us a clearer idea of their effectiveness.