Biofilm Response Research Articles

Identification and comparison of protein composition of biofilms in response to EGCG from Enterococcus faecalis and Staphylococcus lugdunensis, which showed opposite patterns in biofilm-forming abilities.

Bacterial biofilm is resistant to conventional antibiotic treatments, leading to complications associated with many infection-related human diseases. Epigallocatechin Gallate (EGCG), a phenolic catechin enriched in green tea, is recognized for its anti-bacterial and anti-biofilm activities. In this study, we examined the protein components of the biofilms formed in the absence or presence of EGCG using Enterococcus faecalis and Staphylococcus lugdunensis, which had shown opposing patterns in biofilm formation. A clustering heatmap revealed that the two microorganisms expressed the different protein sets in response to EGCG. Proteins that were noticeably upregulated included those associated with stress responsiveness and gluconeogenesis in E. faecalis, and gene modification in S. lugdunensis. Conversely, downregulated proteins were related to tRNA-modifying enzyme activity in E. faecalis, and anabolic metabolism in S. lugdunensis. Among the proteins identified only in EGCG-responsive biofilms, enzymes involved in de novo purine biosynthesis were enriched in E. faecalis, while proteins likely to cause DNA instability and pathogenicity changes were abundantly present in S. lugdunensis. The classification based on gene ontology (GO) terms by microorganism exhibited that metabolic process or catabolic activity was at the top rank in E. faecalis with more than 33 proteins, and in S. lugdunensis, localization or transport was highly ranked with 4 proteins. These results support the hypothesis that EGCG might cause different cellular programs in each microorganism. Finally, comparison of the proteomes between two groups that form biofilms to similar extents discovered that 2 proteins were commonly found in the weak biofilm-forming groups (E. faecalis and EGCG-responding S. lugudunensis), whereas 9 proteins were common among the strong biofilm-forming groups (S. lugdunensis and EGCG-responding E. faecalis). It was suggested that these proteins could serve as potential indicators to detect the presence and predict the extent of biofilm formation by multiple microorganisms. Taken all together, proteomics data and analyses performed in this study provided useful and new information on the proteins embedded in the biofilms formed at the specific conditions, which can aid in diagnosis and the development of tailored treatment strategies.

Improve anti-biofilm efficacy of ultrasound by modulating the phase transition of exopolysaccharides

This study focused on the adverse sonochemical effect of ultrasound on biofilm extracellular polysaccharide and the adaptive biofilm responses for ultrasound resistance. Results showed ultrasound triggered phase transition of polysaccharides within biofilm from solation to gelation, which induced following biofilm viscoelasticity enhancement, consequential failure of biofilm removal and bacteria killing. Introducing additional cationic polysaccharide, 1.25 % chitosan, inhibited the ultrasound responsive polysaccharides gelation and biofilm viscoelasticity enhancement, exerted synergistic antibacterial (97.40 %) and antibiofilm (96.38 %) effects with 120 W ultrasound combined on S. aureus biofilm, prolonged the preservation time of milk 2.45 times longer compared with ultrasound alone. These findings indicated the possible mechanism and solution to improve ultrasound efficacy on biofilm control and bacteria suppression, exhibit the promising prospect of ultrasound combined strategy in hygiene issues of food and medical industry

Lysozyme-Responsive Hydrogels of Chitosan-Streptomycin Conjugates for the On-Demand Release of Biofilm-Dispersing Enzymes for the Efficient Eradication of Oral Biofilms.

Hydrogels with controlled degradation and sustained antibiofilm activity are promising biomaterials for the treatment of oral infections such as periodontitis or caries. In this article, an in situ forming chitosan-streptomycin hydrogel is developed that can target established bacterial biofilms in response to lysozyme, an enzyme that is overexpressed in saliva during oral infections. When the new hydrogel is applied to simulated oral biofilms, the overexpressed lysozyme degrades the hydrogel and releases chitosan-streptomycin oligosaccharides that can eradicate the biofilm. This work has shown that the coupling of chitosan and streptomycin can have a synergistic effect and that the new hydrogel based on chitosan-streptomycin conjugate can effectively combat biofilms of E. coli, S. aureus, and P. aeruginosa formed in vitro achieving a significant reduction in the biomass of the biofilm and a substantial reduction in the population of viable bacteria in established biofilms. Finally, the CS-Str hydrogel loaded with biofilm-disrupting enzymes, in particular, DNase I and/or DspB, showed a significantly increased ability to reduce the biofilm biomass of P. aeruginosa and S. aureus (by over 84% and up to 92%, respectively), resulting in a drastic reduction in cell viability, which fell below 4% for P. aeruginosa and below 5% for S. aureus.

Mitigation of biocorrosion of X80 carbon steel by a shale microbiome biofilm using a green biocide enhanced by d-amino acids

Microbiologically influenced corrosion (MIC) in shale gas field is a major threat with the hydraulic fracturing fluid injected into the subsurface. In this study, the microbiome collected from a shale gas produced water sample was extracted and cultivated in ATCC 1249 medium modified with 10 g/L NaCl anaerobically at 30 °C. d-amino acids, which were reported as biocide enhancers, were found to enhance 2,2-dibromo-3-nitrilopropionamide (DBNPA) biocide on the mitigation of shale microbiome MIC on X80 carbon steel. The combination of 50 ppm (w/w) d-leucine + 50 ppm d-alanine + 1 ppm d-tyrosine had the best enhancement effect on 50 ppm DBNPA with 84 % less weight loss, and 67 % lower corrosion current density (icorr) compared to 50 ppm DBNPA alone. The corrosion data were consistent with the enhanced biofilm inhibition observation. The experimental data also indicated that d-tyrosine used alone at a low dosage of 1 ppm enhanced DBNPA considerably, with 44 % less weight loss and 47 % less icorr. The electrochemical results showed the positive response of shale gas microbiome biofilm to the injected magnetite nanoparticles indicating the extracellular electron transfer might be a main mechanism for its corrosion.

Soluble epoxide hydrolase inhibition impairs triggering receptor expressed on myeloid cells-1 in periodontal tissue.

Periodontitis is a prevalent inflammatory disorder affecting the oral cavity, driven by dysbiotic oral biofilm and host immune response interactions. While the major clinical focus of periodontitis treatment is currently controlling oral biofilm, understanding the immune response is crucial to prevent disease progression. Soluble epoxide hydrolase (sEH) inhibition has shown promise in preventing alveolar bone resorption. Triggering receptors expressed on myeloid cells (TREMs) play pivotal roles in regulating inflammation and bone homeostasis, and dysregulation of TREM signaling is implicated in periodontitis. Here, we investigated the impact of sEH inhibition on TREM 1 and 2 expression, associated with inflammatory cytokines, and histologically assessed the inflammatory infiltrate in periodontal tissue. The experimental periodontitis model was induced by placing a ligature around the upper second molar. For 14 days, animals were treated daily with a sEH inhibitor (TPPU) or vehicle. The alveolar bone loss was examined using a methylene blue stain. Gingival tissues were used to measure the mRNA expression of TREM-1, TREM-2, IKKβ, NF-κB, IL-1β, IL-6, IL-8, and TNF-α by RT-qPCR. Another set of experiments was performed to determine the histological inflammatory scores. In a ligature-induced periodontitis model, sEH inhibition prevented alveolar bone loss and reduced TREM1 expression, albeit with a slight elevation compared to the disease-free group. In contrast, TREM2 expression remained elevated, suggesting sustained immunomodulation favoring resolution. The inhibition of sEH reduced the expression of NF-κB, IL-1β, and TNF-α, while no differences were found in the expression of IL-6, IL-8, and IKKβ. In histological analysis, sEH inhibition reduced the inflammatory leukocyte infiltrate in periodontal tissues close to the ligature. These findings underscore the potential of sEH inhibition to modulate periodontal inflammation by regulating TREM-1 alongside decreased IL-1β and TNF-α expression, highlighting a promising therapeutic approach for periodontitis management.

Responses of suspended sludge and biofilm in a SNAD system under C/N elevation: Microbial activity, nitrogen conversion flux and molecular ecological network

The simultaneous partial nitrification, anammox and denitrification (SNAD) process had received widespread attention as an advanced wastewater treatment process. In this study, the SNAD mainstream nitrogen removal process with the incorporation of polyurethane sponge packing under different C/N conditions was investigated. Results showed that the highest nitrogen removal efficiency of the system was achieved at the C/N of 2.0, while the high C/N (3.5) significantly deteriorate the nitrogen removal efficiency. Meanwhile, high C/N (3.5) significantly inhibited the activity and abundance of anammox bacteria (mainly Candidatus_Kuenenia), resulting in the decreased contribution of anammox (from 63.14 % to 48.09 %). The significant divergence of microbial interactions in the suspended sludge and biofilm was observed with increasing C/N. Compared with suspended sludge, biofilm facilitated higher abundance and activity of anammox bacteria, and the molecular ecological network of biofilm displayed better stability and more efficient mass transfer efficiency between microorganisms. The C/N of 3.5 simplified the subnetworks of Chloroflexi and Proteobacteria but increased the positive interactions between Planctomycetota and other microbes. Anammox bacteria were found as keystone species only in biofilm system. This study provided a theoretical basis and technical guidance for the application of SNAD process in municipal wastewater treatment.

Plastic film from the source of anaerobic digestion: Surface degradation, biofilm and UV response characteristics

This study simulates a major environmental scenario involving “organic fertilizer source” plastics, by exploring the key factors influencing the changes in plastic-films during anaerobic digestion (AD), as well as the responses of the anaerobically digested plastics to ultraviolet (UV) radiation exposure. The results demonstrate that the degradation effect of AD on plastics is reflected by their yellowish and ruptured appearance, slightly worn surfaces, hardening and opacity, and fragmentation. AD significantly increases the content of oxygen-containing functional groups and the degree of unsaturation in plastic films, with thermophilic temperature processes proving more effective than those conducted at mesophilic temperatures. Exposure to UV light has been found to amplify the degradative effects, suggesting the potential cumulative impact of AD and UV. Both AD and UV irradiation reduced the hydrophilicity of plastics. In particular, the hydrophobicity of polylactic acid films was completely disrupted under overlay-exposure. Furthermore, microbial populations on plastic surfaces were mainly bacterial. These bacterial populations were primarily influenced by temperature, and moderately by the plastic types. In contrast, archaea were predominantly affected by both temperature and digested substrate. This study offers a theoretical foundation for strategies aimed at preventing and controlling plastic pollution derived from organic fertilizers.

Unravelling the mechanisms of PFAS toxicity to submerged macrophytes and epiphytic biofilms at metabolic and molecular levels

Per- and poly-fluoroalkyl substances (PFAS) are an emerging class of persistent organic pollutants that are widespread in aquatic ecosystems and pose a serious threat to aquatic organisms. It is thus crucial to explore the toxicity mechanisms of PFAS to submerged macrophytes and biofilms. In this study, Vallisneria natans (V. natans) was exposed to environmentally relevant concentrations of perfluorooctanoic acid (PFOA) and perfluorooctane sulphonate (PFOS). Results showed that PFAS induced the excessive production of reactive oxygen species, triggering antioxidant responses. V. natans exhibited an improved stress tolerance by altering the biosynthesis of several plant secondary metabolites and the histidine, arginine, proline pathways in response to PFAS exposure. Moreover, PIP1–1, PIP2–2, SLAH1 and SLAH2 genes were upregulated, indicating the activation of aquaporins and slow-type anion channels. The uptake of PFOA and PFOS by V. natans was 41.74 % and 52.31 %, respectively. Notably, PFAS bound to functional proteins (GSTF10), promoting the detoxification of plants. Exposure to PFAS also altered the structure of biofilms by inducing the synthesis of large amounts of polysaccharides and proteins. The diversity and richness of the microbial community within periphytic biofilms changed significantly. These results provide a comprehensive description of the responses of aquatic plants and periphytic biofilms to PFAS and the removal mechanism of PFAS, contributing to the environmental risk assessments and removal of PFAS in aquatic ecosystems.

Elemental composition of household dusts extracted in simulated body fluids and their impact on culturable pathogenic bacteria responses

In the last decade, a great deal of research has focused on the determination of potential toxic elements by total concentration and identification the microorganisms in dust. However, determining bio-relevant (e.g., inhalable) forms of elements instead of total contents in acids is necessary for human health. Moreover, examination of the behavior of microorganism under these bio-relevant conditions and revealing the interaction between elements and pathogens is vital and necessary for deeper understanding. However, previous studies have ignored these topics. Therefore, the present study aimed to (i) investigate elements in household dusts extracted in simulated lung fluids, (ii) examine the total concentration of culturable bacteria and their biochemical responses with exposure to bio-fractions of household dusts, and (iii) assess their relations and risks using the model approaches by inhalation. Here, settled dusts were collected in 25 houses, and extracted in four simulated body fluids to determine bio-fractions of elements. Moreover, total count of potentially pathogenic and heterotrophic bacteria, and four clinically important culturable pathogens were incubated in the presence of household-dusts extracted in simulated body fluids. The activity, biofilm, biochemical and oxidative responses of pathogens were measured following household-dust exposures. Afterward, the relationship between elements and pathogen responses were evaluated, and model and derived approaches were used for risk assessments of elements and pathogens. The higher daily intake of elements obtained in artificial lysosomal fluid fraction of household dust mimicking the inflammatory condition compared to other body fluids. Moreover, bacterial responses were mainly influenced from bio-fractions of household dusts and their elemental contents.

Estimating biofilm activity on biofilter elements in recirculating aquaculture systems (RAS) for rearing Atlantic salmon parr (Salmo salar) during operation with ozone and peracetic acid

Chemical disinfection in a recirculating aquaculture system (RAS) may affect biofilm-associated bacteria and the nitrification performance in the biofilter units. The biofilm response to chemical disinfectants in RAS remains unclear, but it can be understood using methods to quantify biofilm activity. Here, we compared the effects of two disinfection strategies, continuous ozone at 0.06 mg/L Cl2 equivalent (Ozone group) and peracetic acid (PAA) at 1 mg/L (PAA group) to control group without disinfectant, on biofilm activity on biofilter elements from nine identical experimental RAS with Atlantic salmon parr (Salmo salar) during a four-week trial. Biofilm activities on biofilter elements from the three groups were examined by measuring oxygen consumption rates (OCR) following sequential spiking with pure tap water, and tap water spiked with nitrite or ammonium, as well as oxygen release rate (kor) following hydrogen peroxide (H2O2) addition, to estimate metabolic activities of biofilm related to endogenous respiration and substrate turnover. The results show that the applied PAA dose increased the endogenous respiration activity of biofilm by 39–133%, stimulated the rate of biofilm enzymatic decomposition of H2O2 by 135%, and partially impaired the biofilm metabolism for nitrite oxidation (decrease by 36%), resulting in a significant nitrite accumulation (rise by 59%) in the cultured water, compared to control over experimental period. Ozone treatment caused an enhanced endogenous respiration activity of biofilm at the beginning of the experiment (increase by 45–74%), but dropped to control levels at the end of the experiment. The results indicate that chronic exposure to PAA can alter the metabolic state of biofilm, which can have consequences for biofilter functions, while chronic exposure to ozone improved water clarity without compromising the metabolic status of biofilm. The investigations provided insights into biofilm response to chemical disinfectant in RAS, which would benefit the optimization of an effective and safe use of disinfectants in RAS.

Environmental health in the upper-middle Luján River basin from a multi-biomarker approach

The objectives of this study were to evaluate the ecophysiological state of the biota using a set of biomarkers in the upper-middle Luján River. To this aim, we collected adult Cnesterodon decemmaculatus fish, biofilm and water at three sampling sites in the upper-middle Luján River (S1: rural area, S2: Luján City and S3: urban area after passing Lujan City). For each site we determined physicochemical variables, heavy metal concentration in water, 19 biomarkers in fish (morphometric, histological, genotoxic, oxidative stress, metabolic and neurotoxic) and six biomarkers in biofilm (oxidative stress and extracellular enzyme). Additionally, we compared the responses of fish and biofilm with those of laboratory controls obtained from outdoor cultures. Our results indicated increased heavy metal concentration at all sites, mainly As and Cd, and decreased dissolved oxygen at S1 and S3. In fish, genotoxic biomarkers showed significant differences with respect to the control. The comet assay indicated damage in fish at the urbanized sites (S2 and S3) and an increased frequency of erythrocytes with nuclear aberrations at all sites. The CEA index (cellular energy allocation), calculated from the metabolic biomarkers and lipid concentration were significantly increased at S1. The gill damage evaluated histologically and with three indices indicated severe damage at all sites. Gills showed thickened primary and secondary lamellae and fusion of filaments at all sites, but a significant increase in mucous cells was only found at S1 and S3. Biofilm showed increased values of extracellular enzymes (β-glucosidase and alkaline phosphatase, lipid peroxidation and oxidative stress enzymes (i.e., catalase) at S3. These results are novel in that they incorporated laboratory controls allowing for comparisons with fish and biofilm from the field. They provided information on the status of a fish population and biofilm community, indicating the negative effect of river water deterioration on the tested organisms. Moreover, results showed what biomarkers were most sensitive for each biological sample.

Response of hyporheic biofilms to temperature changes and dissolved organic carbon enrichment: a mesocosm study

PurposeHyporheic biofilms are the central site for biogeochemical cycling in streams and rivers. In view of global warming and increasing human pressures, this study aimed to compare the response of hyporheic biofilm biomass and activities from an unpolluted reference stream reach surrounded by forest with those from a stream reach exposed to agricultural and urban land use using a mesocosm experiment in which the water temperature and dissolved organic carbon (DOC) contents were manipulated.MethodsHyporheic sediments collected in the field from the two study reaches (i.e. reference and impacted) were incubated in the laboratory at two different temperatures (10 °C, 14 °C) and wetted with three types of synthetic water (control [C] – 0 mg L−1; low DOC – 5 mg L−1; high DOC – 30 mg L−1) for four weeks. The responses of the hyporheic biofilms were measured weekly using structural (total protein content [TPC] as a proxy for biofilm biomass) and functional measures (electron transport system activity [ETSA] and community-level physiological profiling [CLPP]).ResultsThe response of hyporheic biofilms to temperature changes and DOC enrichment was site-specific for all studied measures (TPC, ETSA and CLPP, including measured average well colour development [AWCD]). The addition of DOC to biofilms from the pristine stream reach significantly heightened the responses at 10 °C, a temperature within the normal environmental temperature ranges of the reference location, but not at 14 °C, which was here, a temperature outside normal environmental range. On the other hand, biofilms from the impacted stream reach exhibited increased responses following DOC enrichment under both temperature regimes, with a particularly pronounced response at 14 ºC, in this case, both experimental temperatures were within the normal environmental temperature ranges of the study locations.ConclusionHyporheic biofilms were shown to be, like benthic biofilms, sensitive to temperature changes and organic enrichment, but their response to temperature changes and enrichment caused by climate change and/or other anthropogenic pressures (i.e. point and non-point pollution, removal of the riparian zone, hydromorphological modifications, etc.) was not simply linear but site-specific. The intensity of the response, characterized by increased activity and biomass production, appears to be constrained within the temperature ranges prevalent in the environment from which the biofilms originate. These findings emphasize the importance of site-specific considerations in predicting the impacts of climate change and anthropogenic pressures on these critical components of river and stream ecosystems.

Varying removal of emerging organic contaminants in drinking water treatment residue-based biofilter systems: Influence of hydraulic loading rates on microbiology

Drinking water treatment residue (DWTR)-based biofilter is an environment-friendly and cost-effective technology for eliminating nutrients and emerging organic contaminants (EOCs) from natural water. This study explored the removal efficiencies of five trace-level EOCs and the associated microbial response of biofilm in two identical DWTR-based biofilters under varying hydraulic loading rate (HLR) conditions over six months of operation. The results showed that the most recalcitrant EOC (carbamazepine) was mainly removed by DWTR adsorption (7.8–11.0 ng g−1), while the response of the biofilm community to HLR significantly affected the degradation of 17α-ethinylestradiol, sulfamethoxazole, roxithromycin, and sulfathiazole (P < 0.001). A higher HLR significantly stimulated the production of extracellular polymeric substance and increased the stochasticity and diversity of community assembly, improving denitrification and the removal efficiency of DWTR-based biofilm for biodegradable roxithromycin (80 %) and sulfamethoxazole (76 %). In contrast, lower HLR led to carbon limitation and exerted substrate selection pressures, resulting in more deterministic and stable community assembly, with keystone species such as Bacillus enriched in EOC degradation-related functional genes. Moreover, co-occurrence network analysis revealed a simpler but more intensive interaction network among EOC degraders under the lower HLR condition. This interaction network facilitated the co-metabolism of influent organic carbon and the removal of more recalcitrant contaminants, 17α-ethinylestradiol (48 %) and sulfathiazole (37 %). This study offered a novel insight into biofilter management from a hybrid perspective of environmental microbiology and engineering science, highlighting the dynamic adaptation of microbes to specific EOC degradation under varying HLR conditions.

Dynamic and structural response of a multispecies biofilm to environmental perturbations induced by the continuous increase of benzimidazole fungicides in a permeable reactive biobarrier.

This work explores the dynamics of spatiotemporal changes in the taxonomic structure of biofilms and the degradation kinetics of three imidazole group compounds: carbendazim (CBZ), methyl thiophanate (MT), and benomyl (BN) by a multispecies microbial community attached to a fixed bed horizontal tubular reactor (HTR). This bioreactor mimics a permeable reactive biobarrier, which helps prevent the contamination of water bodies by pesticides in agricultural wastewater. To rapidly quantify the microbial response to crescent loading rates of benzimidazole compounds, a gradient system was used to transiently raise the fungicide volumetric loading rates, measuring the structural and functional dynamics response of a microbial community in terms of the volumetric removal rates of the HTR entering pollutants. The loading rate gradient of benzimidazole compounds severely impacts the spatiotemporal taxonomic structure of the HTR biofilm-forming microbial community. Notable differences with the original structure in HTR stable conditions can be noted after three historical contingencies (CBZ, MT, and BN gradient loading rates). It was evidenced that the microbial community did not return to the composition prior to environmental disturbances; however, the functional similarity of microbial communities after steady state reestablishment was observed. The usefulness of the method of gradual delivery of potentially toxic agents for a microbial community immobilized in a tubular biofilm reactor was shown since its functional and structural dynamics were quickly evaluated in response to fungicide composition and concentration changes. The rapid adjustment of the contaminants' removal rates indicates that even with changes in the taxonomic structure of a microbial community, its functional redundancy favors its adjustment to gradual environmental disturbances.

Structural insights on anti-biofilm mechanism of heated slightly acidic electrolyzed water technology against multi-resistant Staphylococcus aureus biofilm on food contact surface

Slightly acidic electrolyzed water (SAEW) has proven to be an efficient and novel sanitizer in food and agriculture field. This study assessed the efficacy of SAEW (30 mg/L) at 40 °C on the inactivation of foodborne pathogens and detachment of multi-resistant Staphylococcus aureus (MRSA) biofilm. Furthermore, the underlying mechanism of MRSA biofilm under heated SAEW at 40 °C treatment on metabolic profiles was investigated. The results showed that the heated SAEW at 40 °C significantly effectively against foodborne pathogens of 1.96–7.56 (lg (CFU/g)) reduction in pork, chicken, spinach, and lettuce. The heated SAEW at 40 °C treatment significantly reduced MRSA biofilm cells by 2.41 (lg (CFU/cm2)). The synergistic effect of SAEW treatment showed intense anti-biofilm activity in decreasing cell density and impairing biofilm cell membranes. Global metabolic response of MRSA biofilms, treated by SAEW at 40 °C, revealed the alterations of intracellular metabolites, including amino acids, organic acid, fatty acid, and lipid. Moreover, signaling pathways involved in amino acid metabolism, energy metabolism, nucleotide synthesis, carbohydrate metabolites, and lipid biosynthesis were functionally disrupted by the SAEW at 40 °C treatment. As per our knowledge, this is the first research to uncover the potential mechanism of heated SAEW treatment against MRSA biofilm on food contact surface.

The interplay of hematite and photic biofilm triggers the acceleration of biotic nitrate removal

The soil-water interface is replete with photic biofilm and iron minerals; however, the potential of how iron minerals promote biotic nitrate removal is still unknown. This study investigates the physiological and ecological responses of photic biofilm to hematite (Fe2O3), in order to explore a practically feasible approach for in-situ nitrate removal. The nitrate removal by photic biofilm was significantly higher in the presence of Fe2O3 (92.5%) compared to the control (82.8%). Results show that the presence of Fe2O3 changed the microbial community composition of the photic biofilm, facilitates the thriving of Magnetospirillum and Pseudomonas, and promotes the growth of photic biofilm represented by the extracellular polymeric substance (EPS) and the content of chlorophyll. The presence of Fe2O3 also induces oxidative stress (•O2−) in the photic biofilm, which was demonstrated by electron spin resonance spectrometry. However, the photic biofilm could improve the EPS productivity to prevent the entrance of Fe2O3 to cells in the biofilm matrix and mitigate oxidative stress. The Fe2O3 then promoted the relative abundance of Magnetospirillum and Pseudomonas and the activity of nitrate reductase, which accelerates nitrate reduction by the photic biofilm. This study provides an insight into the interaction between iron minerals and photic biofilm and demonstrates the possibility of combining biotic and abiotic methods to improve the in-situ nitrate removal rate.

Insights into the response of electroactive biofilm with petroleum hydrocarbons degradation ability to quorum sensing signals

Bioelectrochemical technologies based on electroactive biofilms (EAB) are promising for petroleum hydrocarbons (PHs) remediation as anode can serve as inexhaustible electron acceptor. However, the toxicity of PHs might inhibit the formation and function of EABs. Quorum sensing (QS) is ideal for boosting the performance of EABs, but its potential effects on reshaping microbial composition of EABs in treating PHs are poorly understood. Herein, two AHL signals, C4-HSL and C12-HSL, were employed to promote EABs for PHs degradation. The start-times of AHL-mediated EABs decreased by 18–26%, and maximum current densities increased by 28–63%. Meanwhile, the removal of total PHs increased to over 90%. AHLs facilitate thicker and more compact biofilm as well as higher viability. AHLs enhanced the electroactivity and direct electron transfer capability. The total abundance of PH-degrading bacteria increased from 52.05% to 75.33% and 72.02%, and the proportion of electroactive bacteria increased from 26.14% to 62.72% and 63.30% for MFC-C4 and MFC-C12. Microbial networks became more complex, aggregated, and stable with addition of AHLs. Furthermore, AHL-stimulated EABs showed higher abundance of genes related to PHs degradation. This work advanced our understanding of AHL-mediated QS in maintaining the stable function of microbial communities in the biodegradation process of petroleum hydrocarbons.

Optimization of periodic treatment strategies for bacterial biofilms using an agent-based in silico approach.

Biofilms are responsible for most chronic infections and are highly resistant to antibiotic treatments. Previous studies have demonstrated that periodic dosing of antibiotics can help sensitize persistent subpopulations and reduce the overall dosage required for treatment. Because the dynamics and mechanisms of biofilm growth and the formation of persister cells are diverse and are affected by environmental conditions, it remains a challenge to design optimal periodic dosing regimens. Here, we develop a computational agent-based model to streamline this process and determine key parameters for effective treatment. We used our model to test a broad range of persistence switching dynamics and found that if periodic antibiotic dosing was tuned to biofilm dynamics, the dose required for effective treatment could be reduced by nearly 77%. The biofilm architecture and its response to antibiotics were found to depend on the dynamics of persister cells. Despite some differences in the response of biofilm governed by different persister switching rates, we found that a general optimized periodic treatment was still effective in significantly reducing the required antibiotic dose. As persistence becomes better quantified and understood, our model has the potential to act as a foundation for more effective strategies to target bacterial infections.

New sensitive tools to characterize meta-metabolome response to short- and long-term cobalt exposure in dynamic river biofilm communities

Untargeted metabolomics is a non-a priori analysis of biomolecules that characterizes the metabolome variations induced by short- and long-term exposures to stressors. Even if the metabolite annotation remains lacunar due to database gaps, the global metabolomic fingerprint allows for trend analyses of dose-response curves for hundreds of cellular metabolites. Analysis of dose/time-response curve trends (biphasic or monotonic) of untargeted metabolomic features would thus allow the use of all the chemical signals obtained in order to determine stress levels (defense or damage) in organisms. To develop this approach in a context of time-dependent microbial community changes, mature river biofilms were exposed for 1 month to four cobalt (Co) concentrations (from background concentration to 1 × 10−6 M) in an open system of artificial streams. The meta-metabolomic response of biofilms was compared against a multitude of biological parameters (including bioaccumulation, biomass, chlorophyll a content, composition and structure of prokaryotic and eukaryotic communities) monitored at set exposure times (from 1 h to 28 d). Cobalt exposure induced extremely rapid responses of the meta-metabolome, with time range inducing defense responses (TRIDeR) of around 10 s, and time range inducing damage responses (TRIDaR) of several hours. Even in biofilms whose structure had been altered by Co bioaccumulation (reduced biomass, chlorophyll a contents and changes in the composition and diversity of prokaryotic and eukaryotic communities), concentration range inducing defense responses (CRIDeR) with similar initiation thresholds (1.41 ± 0.77 × 10−10 M Co2+ added in the exposure medium) were set up at the meta-metabolome level at every time point. In contrast, the concentration range inducing damage responses (CRIDaR) initiation thresholds increased by 10 times in long-term Co exposed biofilms. The present study demonstrates that defense and damage responses of biofilm meta-metabolome exposed to Co are rapidly and sustainably impacted, even within tolerant and resistant microbial communities.

Glycerol metabolism impacts biofilm phenotypes and virulence in Pseudomonas aeruginosa via the Entner-Doudoroff pathway.

Pseudomonas aeruginosa is a ubiquitous bacterium and a notorious opportunistic pathogen that forms biofilm structures in response to many environmental cues. Biofilm formation includes attachment to surfaces and the production of the exopolysaccharide Pel, which is present in both the PAO1 and PA14 laboratory strains of P. aeruginosa. Biofilms help protect bacterial cells from host defenses and antibiotics and abet infection. The carbon source used by the cells also influences biofilm, but these effects have not been deeply studied. We show here that glycerol, which can be liberated from host surfactants during infection, encourages surface attachment and magnifies colony morphology differences. We find that glycerol kinase is important but not essential for glycerol utilization and relatively unimportant for biofilm behaviors. Among downstream enzymes predicted to take part in glycerol utilization, Edd stood out as being important for glycerol utilization and for enhanced biofilm phenotypes in the presence of glycerol. Thus, gluconeogenesis and catabolism of anabolically produced glucose appear to impact not only the utilization of glycerol but also glycerol-stimulated biofilm phenotypes. Finally, waxworm moth larvae and nematode infection models reveal that interruption of the Entner-Doudoroff pathway, but not abrogation of glycerol phosphorylation, unexpectedly increases P. aeruginosa lethality in both acute and chronic infections, even while stimulating a stronger immune response by Caenorhabditis elegans.IMPORTANCEPseudomonas aeruginosa, the ubiquitous environmental bacterium and human pathogen, forms multicellular communities known as biofilms in response to various stimuli. We find that glycerol, a common carbon source that bacteria can use for energy and biosynthesis, encourages biofilm behaviors such as surface attachment and colony wrinkling by P. aeruginosa. Glycerol can be derived from surfactants that are present in the human lungs, a common infection site. Glycerol-stimulated biofilm phenotypes do not depend on phosphorylation of glycerol but are surprisingly impacted by a glucose breakdown pathway, suggesting that it is glycerol utilization, and not its mere presence or cellular import, that stimulates biofilm phenotypes. Moreover, the same mutations that block glycerol-stimulated biofilm phenotypes also impact P. aeruginosa virulence in both acute and chronic animal models. Notably, a glucose-breakdown mutant (Δedd) counteracts biofilm phenotypes but shows enhanced virulence and stimulates a stronger immune response in Caenorhabditis elegans.