Dense Biofilm Research Articles

Desulfovibrio fairfieldensis adhesion on implantable titanium used in odontology: a preliminary study.

The study presented here aimed to assess the ability of Desulfovibrio fairfieldensis bacteria to adhere to and form biofilm on the structure of titanium used in implants. D. fairfieldensis was found in the periodontal pockets in the oral environment, indicating that these bacteria can colonize the implant-bone interface and consequently cause bone infection and implant corrosion. Plates of implantable titanium, of which surfaces were characterized by scanning electronic microscopy and Raman spectroscopy, were immersed in several suspensions of D. fairfieldensis cells containing potassium nitrate on the one hand, and artificial saliva or a sulfato-reducing bacterial culture medium on the other hand. Following various incubation timepoints bacteria were counted in different media to determine their doubling time and titanium samples are checked for and determination of the total number of adhered bacteria and biofilm formation. Adhesion of D. fairfieldensis on titanium occurs at rates ranging from 2.105 to 4.6.106 bacteriah-1cm-2 in the first 18 h of incubation on both native and implantable titanium samples. Following that time, the increase in cell numbers per h and cm2 is attributed to growth in adhered bacteria. After 30 days of incubation in a nutrient-rich medium, dense biofilms are observed forming on the implant surface where bacteria became embedded in a layer of polymers D. fairfieldensis is able of adhering to an implantable titanium surface in order to form a biofilm. Further studies are still necessary, however, to assess whether this adhesion still occurs in an environment containing saliva or serum proteins that may alter the implant surface.

Low Concentrations of Chlorhexidine Inhibit the Formation and Structural Integrity of Enzyme-Treated Multispecies Oral Biofilms

The self-produced matrix of biofilms, consisting of extracellular polymeric substances, plays an important role in biofilm adhesion to surfaces and the structural integrity of biofilms. In dentistry, biofilms cause multiple diseases such as caries, periodontitis, and pulpitis. Disruption of these biofilms adhering to dental hard tissues may pose a major challenge since biofilms show higher tolerance to antimicrobials and antibiotics than planktonic cells. In this study, the effect of low concentrations of chlorhexidine (CHX) on enzyme-treated multispecies oral biofilm was investigated in an in vitro model. Six-species biofilms were enzymatically treated by anaerobic growth in a medium containing DNase I and proteinase K. Biofilms were exposed to a low concentration of CHX at defined time points. After 64h, biofilms were either harvested and quantified by cultural analyses or stained for confocal laser scanning microscopy (CLSM) analyses using either Live/Dead kit or different fluorescent dyes. A mixture of YoPro1 and SYTOX™ Green, Fluorescent Brightener 28 (Calcofluor), and SYPRO™ Ruby Protein Gel Stain was used to stain total DNA, exopolysaccharides, and extracellular proteins, respectively. Extracellular DNA (eDNA) was visualized via an indirect immunofluorescence assay (Mouse anti-DNA IgG, Goat anti-Mouse IgG, Streptavidin-Cy3). Overall, the total colony-forming units significantly decreased after combined treatment with a low concentration of CHX and enzymes compared to the group treated with CHX alone (p<0.001). These findings also apply to five species individually (Streptococcus mutans, Streptococcus oralis, Actinomyces oris, Veillonella dispar, and Candida albicans) occurring in the biofilms, with Fusobacterium nucleatum being the only exception. Furthermore, CLSM images showed less dense biofilms and a reduction in cell numbers after combined treatment compared to the group without enzymes. The combination of enzymes capable of disturbing the matrix integrity with antimicrobial agents thus appears to be a promising approach for biofilm disruption and killing.

The Terrestrial Plastisphere: Diversity and Polymer-Colonizing Potential of Plastic-Associated Microbial Communities in Soil.

The concept of a ‘plastisphere microbial community’ arose from research on aquatic plastic debris, while the effect of plastics on microbial communities in soils remains poorly understood. Therefore, we examined the inhabiting microbial communities of two plastic debris ecosystems with regard to their diversity and composition relative to plastic-free soils from the same area using 16S rRNA amplicon sequencing. Furthermore, we studied the plastic-colonizing potential of bacteria originating from both study sites as a measure of surface adhesion to UV-weathered polyethylene (PE) using high-magnification field emission scanning electron microscopy (FESEM). The high plastic content of the soils was associated with a reduced alpha diversity and a significantly different structure of the microbial communities. The presence of plastic debris in soils did not specifically enrich bacteria known to degrade plastic, as suggested by earlier studies, but rather shifted the microbial community towards highly abundant autotrophic bacteria potentially tolerant to hydrophobic environments and known to be important for biocrust formation. The bacterial inoculates from both sites formed dense biofilms on the surface and in micrometer-scale surface cracks of the UV-weathered PE chips after 100 days of in vitro incubation with visible threadlike EPS structures and cross-connections enabling surface adhesion. High-resolution FESEM imaging further indicates that the microbial colonization catalyzed some of the surface degradation of PE. In essence, this study suggests the concept of a ‘terrestrial plastisphere’ as a diverse consortium of microorganisms including autotrophs and other pioneering species paving the way for those members of the consortium that may eventually break down the plastic compounds.

Improved eradication efficacy of a combination of newly identified antimicrobial agents in C. albicans and S. aureus mixed-species biofilm

Candida albicans and Staphylococcus aureus are common human pathogens, frequently isolated independently or co-isolated from bloodstream infections, and able to form dense polymicrobial biofilms on various medical devices resulting in strong resistance to conventionally used antimicrobials. New and innovative approaches are therefore needed to ensure the successful management of biofilm related infections. In this study, a chalcone-based derivative and a polycyclic anthracene-maleimide adduct, previously ascertained by us as inhibitors of C.albicans and S.aureus growths, respectively, were reconsidered in a new perspective by evaluating the efficacy of a combined treatment against a polymicrobial biofilm. Both quantitative and qualitative analyses were carried out to delve into their inhibitory potential on the polymicrobial population. Our results indicate that these newly identified antimicrobials are effective in reducing the biomass of the mixed C.albicans-S. aureus biofilm and the viability of fungal-bacterial cells within the polymicrobial community; in addition, confocal laser scanning microscopy demonstrates that the combined treatment thoroughly modifies the architecture of the dual-species biofilm.

Confocal spectral microscopy, a non-destructive approach to follow contamination and biofilm formation of mCherry Staphylococcus aureus on solid surfaces

Methods to test the safety of wood material for hygienically sensitive places are indirect, destructive and limited to incomplete microbial recovery via swabbing, brushing and elution-based techniques. Therefore, we chose mCherry Staphylococcus aureus as a model bacterium for solid and porous surface contamination. Confocal spectral laser microscope (CSLM) was employed to characterize and use the autofluorescence of Sessile oak (Quercus petraea), Douglas fir (Pseudotsuga menziesii) and poplar (Populus euramericana alba L.) wood discs cut into transversal (RT) and tangential (LT) planes. The red fluorescent area occupied by bacteria was differentiated from that of wood, which represented the bacterial quantification, survival and bio-distribution on surfaces from one hour to one week after inoculation. More bacteria were present near the surface on LT face wood as compared to RT and they persisted throughout the study period. Furthermore, this innovative methodology identified that S. aureus formed a dense biofilm on melamine but not on oak wood in similar inoculation and growth conditions. Conclusively, the endogenous fluorescence of materials and the model bacterium permitted direct quantification of surface contamination by using CSLM and it is a promising tool for hygienic safety evaluation.

Mucosal Biofilms Are an Endoscopic Feature of Irritable Bowel Syndrome and Ulcerative Colitis

Background & AimsIrritable bowel syndrome (IBS) and inflammatory bowel diseases result in a substantial reduction in quality of life and a considerable socioeconomic impact. In IBS, diagnosis and treatment options are limited, but evidence for involvement of the gut microbiome in disease pathophysiology is emerging. Here we analyzed the prevalence of endoscopically visible mucosal biofilms in gastrointestinal disease and associated changes in microbiome composition and metabolism.MethodsThe presence of mucosal biofilms was assessed in 1426 patients at 2 European university-based endoscopy centers. One-hundred and seventeen patients were selected for in-depth molecular and microscopic analysis using 16S ribosomal RNA gene amplicon-sequencing of colonic biopsies and fecal samples, confocal microscopy with deep learning–based image analysis, scanning electron microscopy, metabolomics, and in vitro biofilm formation assays.ResultsBiofilms were present in 57% of patients with IBS and 34% of patients with ulcerative colitis compared with 6% of controls (P < .001). These yellow-green adherent layers of the ileum and right-sided colon were microscopically confirmed to be dense bacterial biofilms. 16S-sequencing links the presence of biofilms to a dysbiotic gut microbiome, including overgrowth of Escherichia coli and Ruminococcus gnavus. R. gnavus isolates cultivated from patient biofilms also formed biofilms in vitro. Metabolomic analysis found an accumulation of bile acids within biofilms that correlated with fecal bile acid excretion, linking this phenotype with a mechanism of diarrhea.ConclusionsThe presence of mucosal biofilms is an endoscopic feature in a subgroup of IBS and ulcerative colitis with disrupted bile acid metabolism and bacterial dysbiosis. They provide novel insight into the pathophysiology of IBS and ulcerative colitis, illustrating that biofilm can be seen as a tipping point in the development of dysbiosis and disease.

Microbial Communities on Plastic Polymers in the Mediterranean Sea.

Plastic particles in the ocean are typically covered with microbial biofilms, but it remains unclear whether distinct microbial communities colonize different polymer types. In this study, we analyzed microbial communities forming biofilms on floating microplastics in a bay of the island of Elba in the Mediterranean Sea. Raman spectroscopy revealed that the plastic particles mainly comprised polyethylene (PE), polypropylene (PP), and polystyrene (PS) of which polyethylene and polypropylene particles were typically brittle and featured cracks. Fluorescence in situ hybridization and imaging by high-resolution microscopy revealed dense microbial biofilms on the polymer surfaces. Amplicon sequencing of the 16S rRNA gene showed that the bacterial communities on all plastic types consisted mainly of the orders Flavobacteriales, Rhodobacterales, Cytophagales, Rickettsiales, Alteromonadales, Chitinophagales, and Oceanospirillales. We found significant differences in the biofilm community composition on PE compared with PP and PS (on OTU and order level), which shows that different microbial communities colonize specific polymer types. Furthermore, the sequencing data also revealed a higher relative abundance of archaeal sequences on PS in comparison with PE or PP. We furthermore found a high occurrence, up to 17% of all sequences, of different hydrocarbon-degrading bacteria on all investigated plastic types. However, their functioning in the plastic-associated biofilm and potential role in plastic degradation needs further assessment.

Local Acidification Limits the Current Production and Biofilm Formation of Shewanella oneidensis MR-1 With Electrospun Anodes.

The anodic current production of Shewanella oneidensis MR-1 is typically lower compared to other electroactive bacteria. The main reason for the low current densities is the poor biofilm growth on most anode materials. We demonstrate that the high current production of Shewanella oneidensis MR-1 with electrospun anodes exhibits a similar threshold current density as dense Geobacter spp biofilms. The threshold current density is a result of local acidification in the biofilm. Increasing buffer concentration from 10 to 40 mM results in a 1.8-fold increase of the current density [(590 ± 25) μA cm−2] while biofilm growth stimulation by riboflavin has little effect on the current production. The current production of a reference material below the threshold did not respond to the increased buffer concentration but could be enhanced by supplemented riboflavin that stimulated the biofilm growth. Our results suggest that the current production with S. oneidensis is limited (1) by the biofilm growth on the anode that can be enhanced by the choice of the electrode material, and (2) by the proton transport through the biofilm and the associated local acidification.

Effect of Pseudomonas aeruginosa Strain ZK Biofilm on the Mechanical and Corrosion Behavior of 316L Stainless Steel and α-brass

This research work aims to investigate the effect of the aerobic bacterium, Pseudomonas aeruginosa on the mechanical and electrochemical properties of the 316L stainless steel and α-brass. These properties of both the alloys were determined after 7 days of exposure to the controlled and inoculated media at 37°C. The microstructural and electrochemical test results revealed the deleterious effects of Pseudomonas aeruginosa. After exposure to the inoculated medium, the scanning electron microscopy (SEM) results showed the larger pitting and formation of relatively dense biofilm on α-brass compared to 316L stainless steel. The tensile strength and hardness of 316L stainless steel were slightly affected after exposure to the controlled and inoculated media. After exposure to the controlled medium and inoculated media, the tensile strength of the α-brass was least affected but a significant decrease in the hardness (from 165 HV to 124 HV) was observed due to the severe attack induced by the Pseudomonas aeruginosa. Similarly, the open-circuit potential of the 316L stainless steel in the inoculated medium was measured to be less active (−410 mV vs Ag/AgCl) than α-brass (−550 mV vs Ag/AgCl). In the inoculated medium, potentiodynamic polarization curves confirmed the severe attack of Pseudomonas aeruginosa on α-brass (7.15 × 10−2 mm/year) compared to 316L stainless steel which registered a corrosion rate of 5.14 × 10−4 mm/year.

Characterization of crude oil degrading bacterial communities and their impact on biofilm formation

In the present study, produced water sample collected from the Indian crude oil reservoir is used to enrich the bacterial communities. The impact of these enriched bacterial communities on the biodegradation of crude oil, biofilm formation, and biocorrosion process are elucidated. A crude oil degradation study is carried out with the minimal salt medium and 94% of crude oil was utilized by enriched bacterial communities. During the crude oil degradation many enzymes including alkane hydroxylase, alcohol dehydrogenase, and lipase are playing a key role in the biodegradation processes. The role of enriched bacterial biofilm on biocorrosion reactions are monitored by weight loss studies and electrochemical analysis. Weight loss study revealed that the biotic system has vigorous corrosion attacks compared to the abiotic system. Both AC-Impedance and Tafel analysis confirmed that the nature of the corrosion reaction take place in the biotic system. Very less charge transfer resistance and higher corrosion current are observed in the biotic system than in the abiotic system. Scanning electron microscope confirms that the dense biofilm formation favoured the pitting type of corrosion. X-ray diffraction analysis confirms that the metal oxides formed in the corrosion systems (biotic). From the metagenomic analysis of the V3–V4 region revealed that presence of diverse bacterial communities in the biofilm, and most of them are uncultured/unknown. Among the known genus, Bacillus, Halomonas, etc are dominant in the enriched bacterial biofilm sample. From this study, we conclude that the uncultured bacterial strains are found to be playing a key role in the pitting type of corrosion and they can utilize crude oil hydrocarbons, which make them succeeded in extreme oil reservoir environments.

Human microbiota modulation via QseC sensor kinase mediated in the Escherichia coli O104:H4 outbreak strain infection in microbiome model

BackgroundThe intestinal microbiota plays a crucial role in human health, adjusting its composition and the microbial metabolites protects the gut against invading microorganisms. Enteroaggregative E. coli (EAEC) is an important diarrheagenic pathogen, which may cause acute or persistent diarrhea (≥14 days). The outbreak strain has the potent Shiga toxin, forms a dense biofilm and communicate via QseBC two-component system regulating the expression of many important virulence factors.ResultsHerein, we investigated the QseC histidine sensor kinase role in the microbiota shift during O104:H4 C227–11 infection in the colonic model SHIME® (Simulator of the Human Intestinal Microbial Ecosystem) and in vivo mice model. The microbiota imbalance caused by C227–11 infection affected ỿ-Proteobacteria and Lactobacillus spp. predominance, with direct alteration in intestinal metabolites driven by microbiota change, such as Short-chain fatty acids (SCFA). However, in the absence of QseC sensor kinase, the microbiota recovery was delayed on day 3 p.i., with change in the intestinal production of SCFA, like an increase in acetate production. The higher predominance of Lactobacillus spp. in the microbiota and significant augmented qseC gene expression levels were also observed during C227–11 mice infection upon intestinal depletion. Novel insights during pathogenic bacteria infection with the intestinal microbiota were observed. The QseC kinase sensor seems to have a role in the microbiota shift during the infectious process by Shiga toxin-producing EAEC C227–11.ConclusionsThe QseC role in C227–11 infection helps to unravel the intestine microbiota modulation and its metabolites during SHIME® and in vivo models, besides they contribute to elucidate bacterial intestinal pathogenesis and the microbiota relationships.

Mixed-culture biocathodes for acetate production from CO2 reduction in the microbial electrosynthesis: Impact of temperature

The temperature effect on bioelectrochemical reduction of CO2 to acetate with a mixed-culture biocathode in the microbial electrosynthesis was explored. The results showed that maximum acetate amount of 525.84 ± 1.55 mg L−1 and fastest acetate formation of 49.21 ± 0.49 mg L−1 d−1 were obtained under mesophilic conditions. Electron recovery efficiency for CO2 reduction to acetate ranged from 14.50 ± 2.20% to 64.86 ± 2.20%, due to propionate, butyrate and H2 generation. Mesophilic conditions were demonstrated to be more favorable for biofilm formation on the cathode, resulting in a stable and dense biofilm. At phylum level, the relative abundance of Bacteroidetes phylum in the biofilm remarkably increased under mesophilic conditions, compared with that at psychrophilic and thermophilic conditions. At genus level, the Clostridium, Treponema, Acidithiobacillus, Acetobacterium and Acetoanaerobium were found to be dominated genera in the biofilm under mesophilic conditions, while genera diversity decreased under psychrophilic and thermophilic conditions.

Microscopic techniques to evaluate the biofilm formation ability of a marine bacterium Pseudomonas aeruginosa PFL-P1 on different substrata.

Biofilm formation in bacteria is strongly affected by the nature of substrata. Different substrata such as glass, polystyrene, steel, ceramic, and rubber were used to assess the biofilm forming ability of a marine bacterium Pseudomonas aeruginosa PFL-P1 using a scanning electron microscope (SEM), atomic force microscope (AFM), and confocal laser scanning microscope (CLSM). The bacterium formed dense biofilms with varied aggregation on different substrata. SEM study revealed small rod-shaped cells with diverse arrangements within the biofilms on all the substrata under study. The AFM study revealed the highest roughness of 545 nm on the ceramic substratum. The biofilms formed on ceramic substratum were characterized with maximum roughness (742 nm), maximum peak height (1,480 nm), and maximum arithmetic mean height (611 nm), significantly higher than all the other substrata (p < .05). AFM studies confirmed that P. aeruginosa PFL-P1 exhibited biofilm heterogeneity on all the substrata. The CLSM study indicated a higher fraction of nucleic acids to α-polysaccharides ratio in the biofilms. COMSTAT analysis revealed the highest biofilm biomass of ~18 μm3 /μm2 on the ceramic substratum. The maximum biofilm thickness of ~50 μm in the native state on the ceramic substratum was significantly higher than glass (p=.0015), polystyrene (p=.0001), steel (p=.0035), and rubber substrata (p=.0001). The higher surface roughness of ceramic substratum is accountable for more area for colonization, as evident from higher biomass and thickness of the biofilm. This study provides insight into the substratum properties, which modulate the biofilm forming ability in bacteria.

Structural and mechanical characterization of biofilm-associated bacterial polymer in the emulsification of petroleum hydrocarbon.

The online version contains supplementary material available at 10.1007/s13205-021-02795-8.

Effects of green synthesized and commercial nZVI on crystal violet degradation by Burkholderia vietnamiensis C09V: Dose-dependent toxicity and biocompatibility

The increasingly common remedial application of nanoscale zero-valent iron (nZVI) to alleviate specific contaminant issues may inadvertently lead to nZVI accumulation in wastewater. This is a potential concern, because the effect of nZVI on the common microbes essential for wastewater biotreatment is not known. This is further complicated when there are many ways available to synthesize nZVI, which may interreact with bacteria differently. Thus, in this study, the different effects of nZVI synthesized by Eucalyptus leaves (EL-nZVI) and a commercially synthesized nZVI on the biodegradation of crystal violet by Burkholderia vietnamiensis C09V (B.V. C09V) was studied. At high dose (1000 mg/L), EL-nZVI and commercial nZVI both significantly inhibited the removal of crystal violet by B.V. C09V, decreasing removal rates by 10.5 and 13.1% respectively. Optical density (OD600) and soluble protein assays indicated that the growth of B.V. C09V improved under low doses (100 mg/L), but remained inhibited under high doses (500 and 1000 mg/L) of both commercial and EL-nZVI. Enzymes were also sensitive to nZVI, where the commercial variant exerted a greater effect on both the activity of lactate dehydrogenase (LDH) and superoxide dismutase (SOD) than EL-nZVI, indicating that EL-nZVI was less toxic than commercial nZVI. LIVE/DEAD staining also showed that the number of apoptotic cells was significantly higher when exposed to commercial nZVI rather than EL-nZVI. Furthermore, scanning electron microscopy (SEM) confirmed that direct contact between nZVI and cells at 1000 mg/L nZVI caused cell membrane disruption. Whereas, at 100 mg/L EL-nZVI, B.V. C09V grew better due to the formation of dense biofilms around the suspended EL-nZVI at a. Fourier transform infrared spectra (FTIR), confirmed an abundance of oxygen-containing functional groups on the surface of EL-nZVI which provided better biocompatibility than commercial nZVI. Overall, while dose was the most significant factor influencing nZVI toxicity, surface composition and morphology was also important. These new findings suggest chemical synthesis of metal nanoparticles should be replaced by biosynthetic routes to maintain viable microbial pollution during wastewater treatment.

Enhanced Eradication of Bacterial/Fungi Biofilms by Glucose Oxidase-Modified Magnetic Nanoparticles as a Potential Treatment for Persistent Endodontic Infections.

Bacterial/fungal biofilm-mediated persistent endodontic infections (PEIs) are one of the most frequent clinical lesions in the oral cavity, resulting in apical periodontitis and tooth damage caused by loss of minerals. The conventional root canal disinfectants are poorly bio-safe and harmful to teeth and tissues, making them ineffective in treating PEIs. The development of nanomaterials is emerging as a promising strategy to eradicate disease-related bacteria/fungi. Herein, glucose oxidase (GOx)-modified magnetic nanoparticles (MNPs) were synthesized via a facile and versatile route for investigating their effects on removing PEI-related bacterial/fungal biofilms. It is found that GOx was successfully immobilized on the MNPs by detecting the changes in the diameter, chemical functional group, charge, and magnetic response. Further, we demonstrate that GOx-modified MNPs (GMNPs) exhibit highly effective antibacterial activity against Enterococcus faecalis and Candida albicans. Moreover, the antibacterial/fungal activity of GMNPs is greatly dependent on their concentrations. Importantly, when placed in contact with bacterial/fungal biofilms, the dense biofilm matrix is destructed due to the movement of GMNPs induced by the magnetic field, the formation of reactive oxygen species, and nutrient starvation induced by GOx. Also, the in vitro experiment shows that the as-prepared GMNPs have excellent cytocompatibility and blood compatibility. Thus, GMNPs offer a novel strategy to treat bacteria/fungi-associated PEIs for potential clinical applications.

Detection of Bacterial Contamination in Dental Unit Water Lines and Testing the Effectiveness of Disinfectants against these Contaminants

Aim: Contaminated dental unit water lines (DUWLs) are a possible source for spreading microorganisms in dental practices. The aim of this study was to detect the bacterial contamination of dental unit water and investigate the effectiveness of the disinfectants.&#x0D; Methodology: Bacterial contamination was detected by a) using bacterial culture of heterotrophic bacteria, total coliforms and Pseudomonas aeruginosa and b) Scanning electron microscopy (SEM) of DUWLs tube. Subsequently, dentists were suggested to treat the DUWLs with disinfectants to eradicate bacterial contaminants and its effectiveness was tested after three months.&#x0D; Results: Bacterial contamination of the water samples ranged from not detected to 2.38×106 CFU/mL. Out of 34 DUWLs water samples tested, 30 (88.24%) samples exceeded the Centers for Disease Control and Prevention (CDC) and Environmental Protection Agency (EPA) recommended threshold of ≤500 CFU /mL, whereas only 4 (11.76%) samples met the standards. Contamination by total coliforms and P. aeruginosa was detected in 52.94% and 64.7% of samples respectively. SEM displayed a dense biofilm on DUWLs tubing confirming the bacterial contamination. The intervention for disinfection of DUWLs resulted more than 50% samples with acceptable bacterial count in test performed after three months.&#x0D; Conclusion: The high rate of bacterial contamination of dental unit water highlights the need to disinfect and monitor the quality of DUWLs periodically.

Non-Invasive Single-Cell Morphometry in Living Bacterial Biofilms

In recent years, fluorescence light sheet-based imaging approaches have been developed to overcome the drawbacks of confocal microscopy, namely phototoxicity and photobleaching. Among these, lattice light sheet microscopy (LLSM) combines excellent 3D spatial resolution with fast temporal resolution and low phototoxicity at levels that cannot be matched by confocal microscopy. However, this potential has not yet been fully realized for investigations of individual cell behaviors and phenotypic changes in dense 3D bacterial biofilms.

Magneto-Based Synergetic Therapy for Implant-Associated Infections via Biofilm Disruption and Innate Immunity Regulation.

Implant‐associated infections (IAIs) are a common cause of orthopedic surgery failure due to microbial biofilm‐induced antibiotic‐resistance and innate immune inactivation. Thus, the destruction of microbial biofilm plays a key role in reducing IAIs. Herein, first, a magneto‐based synergetic therapy (MST) is proposed and demonstrated against IAIs based on biofilm destruction. Under an alternating magnetic field (AMF), CoFe2O4@MnFe2O4 nanoparticles (MNPs), with a rather strong magnetic hyperthermal capacity, can generate sufficient thermal effect to cause dense biofilm dispersal. Loosened biofilms provide channels through which nitrosothiol‐coated MNPs (MNP‐SNOs) can penetrate. Subsequently, thermosensitive nitrosothiols rapidly release nitric oxide (NO) inside biofilms, thus efficiently killing sessile bacteria under the magnetothermal effect of MNPs. More importantly, MNP‐SNOs can trigger macrophage‐related immunity to prevent the relapse of IAIs by exposing the infected foci to a consistent innate immunomodulatory effect. The notable anti‐infection effect of this nanoplatform is also confirmed in a rat IAI model. This work presents the promising potential of combining magnetothermal therapy with immunotherapy, for the effective and durable control and elimination of IAIs.

Attenuation of quorum sensing mediated virulence factors production and biofilm formation in Pseudomonas aeruginosa PAO1 by Colletotrichum gloeosporioides HM3

Signal dependent microbial communication in Pseudomonas aeruginosa PAO1 is a typical phenomenon mediated by acyl homo-serine lactone molecules that helps in developing biofilm and enhance antibiotic resistance. Microbial sources provide insight to the hidden treasure of secondary metabolites, and these structurally diversified chemical motifs can be used as antimicrobial and anti-infective agents. In the present study, endophytic fungus, Colletotrichum gloeosporioides HM3 isolated from Carica papaya leaves was explored for anti-infective potential against P. aeruginosa PAO1. The crude extract of C. gloeosporioides HM3 displayed bacteriostatic effect on P. aeruginosa PAO1 growth at 750 μg/ml concentration. A significant decline was observed in the production of quorum sensing regulated virulence factors, i.e. 56.32%, 62.54%, and 66.67% of pyocyanin, chitinase, and elastase enzyme, respectively. A drastic reduction in pathogenic determinant behaviour after treatment with crude extract of C. gloeosporioides HM3 i.e. EPS, rhamnolipid, and HCN production was noted. Light microscopy and CLSM analysis revealed that fungal extract treatment has reduced bacterial ability to form dense biofilm architecture. In silico analysis demonstrated the binding efficiency of bioactive compound, 4-(2,3-dimethoxybenzylidene)-3-methyl-1-(4-nitrophenyl)-2-pyrazolin-5-one, which is equipotent to the natural ligand and displayed a docking score of −5.436 kcal/mol with QS transcriptional regulator (LasR). Whereas the compound Acetamide, n-[tetrahydro-3-(phenylmethyl) thieno [3,4-d]thiazol-2 (3 h)-ylidene]-, s,s-dioxide exhibits a docking score of −4.088 kcal/mol (LasR) and −1.868 kcal/mol (RhlR) with cognate receptor proteins. Henceforth, the research report suggests C. gloeosporioides HM3 derived metabolites could be considered as a potential inhibitors of QS regulated virulence factors and biofilm production in P. aeruginosa PAO1.