Sessile Cells Research Articles

Proteomic Analysis of Listeria monocytogenes FBUNT During Biofilm Formation at 10°C in Response to Lactocin AL705.

Listeria monocytogenes is one of the major food-related pathogens and is able to survive and multiply under different stress conditions. Its persistence in industrial premises and foods is partially due to its ability to form biofilm. Thus, as a natural strategy to overcome L. monocytogenes biofilm formation, the treatment with lactocin AL705 using a sublethal dose (20AU/ml) was explored. The effect of the presence of the bacteriocin on the biofilm formation at 10°C of L. monocytogenes FBUNT was evaluated for its proteome and compared to the proteomes of planktonic and sessile cells grown at 10°C in the absence of lactocin. Compared to planktonic cells, adaptation of sessile cells during cold stress involved protein abundance shifts associated with ribosomes function and biogenesis, cell membrane functionality, carbohydrate and amino acid metabolism, and transport. When sessile cells were treated with lactocin AL705, proteins’ up-regulation were mostly related to carbohydrate metabolism and nutrient transport in an attempt to compensate for impaired energy generation caused by bacteriocin interacting with the cytoplasmic membrane. Notably, transport systems such as β-glucosidase IIABC (lmo0027), cellobiose (lmo2763), and trehalose (lmo1255) specific PTS proteins were highly overexpressed. In addition, mannose (lmo0098), a specific PTS protein indicating the adaptive response of sessile cells to the bacteriocin, was downregulated as this PTS system acts as a class IIa bacteriocin receptor. A sublethal dose of lactocin AL705 was able to reduce the biofilm formation in L. monocytogenes FBUNT and this bacteriocin induced adaptation mechanisms in treated sessile cells. These results constitute valuable data related to specific proteins targeting the control of L. monocytogenes biofilm upon bacteriocin treatment.

Catechin as the Most Efficient Bioactive Compound from Azadirachta indica with Antibiofilm and Anti-quorum Sensing Activities Against Dental Biofilm: an In Vitro and In Silico Study.

Neem (Azadirachta indica [AI]) is a unique and traditional source of antioxidant and antibacterial compounds. The GC-MS studies revealed that phytoextract of Azadirachta indica comprises a large number of phytocompounds that possess the efficacy of inhibiting the biofilm. It was observed that phytocompounds like catechin showed maximum eradication of biofilm along with the degradation of EPS structural components like carbohydrates and proteins compared to quercetin, nimbolide, nimbin, and azardirachtin, and hence, catechin was proved to be the best against dental plaque-forming bacteria. It was also observed that catechin was able to bring about a marked reduction in quorum sensing (QS) both in Alcaligenes faecalis and Pseudomonas gingivalis dental biofilm-forming strains. The extent of such reduction was maximum for catechin (94.56±2.56% in P. gingivalis & 96.56±2.5 in A. faecalis) in comparison to other bioactive compounds. It was further observed that the bioactive compounds possess the ability to quickly pass across the membrane and bring about inhibition in the DNA and RNA content of the sessile cells. This was further validated by microscopic and in silico studies. Thus, this study revealed that catechin obtained from the phytoextract of AI showed a marked ability to inhibit the dental biofilm and can be used as a natural drug-like compound in treating biofilm-associated chronic infections.

Efficacy of dry sanitizing methods on Listeria monocytogenes biofilms

This study assessed the efficacy of five dry sanitizing treatments on Listeria monocytogenes biofilms grown on two different surface types; stainless steel (SS) and polypropylene (PP). Treatments with 70% ethanol and a commercial product (quaternary ammonium compound + isopropyl alcohol) exhibited similar weak performance on both SS and PP, with the exception of the commercial product on SS at 30 min. An inactivation curve (log N/N0), characterized by two plateaus, was observed on SS after the dry heat treatment (90 °C). However, there was no significant difference (p < 0.05) throughout time. On PP, 30 min of exposure led to a reduction of 3.7 log CFU/cm2. The surface material influenced (p < 0.05) the performance of UV-C light and gaseous ozone. On PP, L. monocytogenes sessile cells were reduced below the limit of detection (1.7 log CFU/cm2) within 5 min for both treatments. On SS, reductions up to 2.2 and 3.4 log CFU/cm2 were observed for UV-C and ozone, respectively. According to the Weibull model, the time required to achieve 5-log reductions (t5d) was quite long (>4.9 × 10 min), with the exception of UV-C and ozone on PP (ca. 6 min). In conclusion, dry heat was the most effective treatment on SS, and gaseous ozone and UV-C on PP.

Interactions between L. monocytogenes and P. fluorescens in Dual-Species Biofilms under Simulated Dairy Processing Conditions.

In dairy processing environments, many bacterial species adhere and form biofilms on surfaces and equipment, leading to foodborne illness and food spoilage. Among them, Listeria monocytogenes and Pseudomonas spp. could be present in mixed-species biofilms. This study aimed to evaluate the interactions between L. monocytogenes and P. fluorescens in biofilms simulating dairy processing conditions, as well as the capability of P. fluorescens in co-culture to produce the blue pigment in a Ricotta-based model system. The biofilm-forming capability of single- and mixed-cultures was evaluated on polystyrene (PS) and stainless steel (SS) surfaces at 12 °C for 168 h. The biofilm biomass was measured, the planktonic and sessile cells and the carbohydrates in biofilms were quantified. The biofilms were also observed through Confocal Laser Scanning Microscopy analysis. Results showed that only P. fluorescens was able to form biofilms on PS. Moreover, in dual-species biofilms at the end of the incubation time (168 h at 12 °C), a lower biomass compared to P. fluorescens mono-species was observed on PS. On SS, the biofilm cell population of L. monocytogenes was higher in the dual-species than in mono-species, particularly after 48 h. Carbohydrates quantity in the dual-species system was higher than in mono-species and was revealed also at 168 h. The production of blue pigment by P. fluorescens was revealed both in single- and co-culture after 72 h of incubation (12 °C). This work highlights the interactions between the two species, under the experimental conditions studied in the present research, which can influence biofilm formation (biomass and sessile cells) but not the capability of P. fluorescens to produce blue pigment.

The Effect of Dual-Species Biofilms, Monosaccharide and D-Amino Acids on Pseudomonal Biofilm

Background: The major therapeutic failure in clinical settings is due to problematic biofilm-producingbacteria like Pseudomonas aeruginosa. This study aims to investigate the effect of dual-species biofilms,monosaccharide, and D-amino acids on pseudomonal biofilm. Methodology: A total of 130 patients withcatheter-associated urinary tract and wound infections were involved in this study. Quantitative biofilmassay by alginate beads was performed. The dual-species biofilms have been done, and the effect ofciprofloxacin, monosaccharides, and D-amino acids on sessile cells was detected. Conclusion: This studysuggested that the combined action of both monosaccharides (glucose and galactose) and the combination ofeach one with ciprofloxacin is the enhancing of P. aeruginosa biofilm and increases survival strategy againstciprofloxacin. In contrast, a noticeable biofilm reduction and marked synergistic action for ciprofloxacinhave been determined in the combination of the two D-amino acid, alanine, and glycine in comparison tothe effect of each one alone. Furthermore, the dual biofilm of P. aeruginosa with each of K. pneumonia andE. coli, markedly reduced pseudomonal biofilm, while co-culture with S. aureus revealed strong support ofpseudomonal pathogenicity and increased its biofilm production.

Study on Biofilm Forming Microorganisms Associated with the Biocorrosion of X80 Pipeline Steel in Produced Water from Oilfield

Abstract Biocorrosion is the main process that causes pipeline damages and losses in the oil industry. The objective of this work was to investigate the influence of biofilm forming microorganisms on the biocorrosion of X80 steel exposed in produced water through microbiological characterization, film and biofilm analysis by optical microscopy and scanning electron microscopy, weight loss and surface analysis by laser confocal microscopy. Changes in produced water after 360 days were attributed to planktonic cells, temperature conditions, contact with air, photo-oxidation, biodegradation, and seasonality. The total aerobic bacteria presented sessile cell concentration of 7.39 x 104 CFU/cm2, while the other investigated groups showed lower concentrations. The micrography of the film showed salt crystals, whereas in the biofilm microorganisms, exopolysaccharides and corrosion products were observed. Weight loss after 360 days for the abiotic and biotic systems was 0.0222 g/cm2 and 0.3039 g/cm2, respectively, showing that microorganisms accentuated the corrosion of X80 steel.

Bacterial adhesion on spinal implants: An in vitro study of "hot spots".

Few studies evaluated bacterial colonization of spinal implants from a "topographic" point of view. This lack of knowledge could hinder the development of more effective strategies in the prevention and treatment of postoperative spinal infections. The aim of this in vitro study was the analysis of the adhesion pattern of sessile cells on conventional spinal implants, to identify "hot spots" on implants where bacterial adhesion could be favored. Clinically relevant Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa isolates were grown on commercially available end product spinal implants. To identify sessile cells attached to implant surfaces, confocal laser scan microscopy was used. Different areas from the spinal instrumentations (both Ti and CoCr) were selected for biofilm quantification. Bacterial biofilm was markedly increased in the cut of the rods, both Ti and CoCr, as the uneven surface deriving from the cut might foster cell adhesion. Though not statistically significant, a difference was observed between the rod and the area of the notch, possibly as a consequence of the smoothening effect deriving from the bending of the rod. Finally, the amount of biofilm produced on cobalt-chromium surfaces was always more significant than that formed on titanium surfaces. This study highlights how bacterial adhesion through biofilm formation is favored on the surfaces of higher irregularity and that staphylococci are able to increase sessile biomass on CoCr surfaces. These preliminary results show how surface modifications on the implants may play a key role in bacterial adhesion, opening an exciting field for future research.

Clinical Biofilm Ring Test® Reveals the Potential Role of β-Lactams in the Induction of Biofilm Formation by P. aeruginosa in Cystic Fibrosis Patients.

Biofilms are characterized by high tolerance to antimicrobials. However, conventional antibiograms are performed on planktonic microorganisms. Through the clinical Biofilm Ring Test® (cBRT), initially aimed to measure the adhesion propensity of bacteria, we discerned a variable distribution of biofilm-producer strains among P. aeruginosa samples isolated from expectorations of cystic fibrosis (CF) patients. Despite a majority of spontaneous adherent isolates, few strains remained planktonic after 5 h of incubation. Their analysis by an adapted protocol of the cBRT revealed an induction of the biofilm early formation by sub-inhibitory doses of β-lactams. Microscopic observations of bacterial cultures stained with Syto 9/Propidium Iodide (PI) confirmed the ability of antimicrobials to increase either the bacterial biomass or the biovolume occupied by induced sessile cells. Finally, the cBRT and its derivatives enabled to highlight in a few hours the potential inducer property of antibiotics on bacterial adhesion. This phenomenon should be considered carefully in the context of CF since patients are constantly under fluctuating antimicrobial treatments. To conclude, assays derived from the Biofilm Ring Test® (BRT) device, not only define efficient doses preventing biofilm formation, but could be useful for the antimicrobial selection in CF, to avoid inducer molecules of the early biofilm initiation.

Sulfate reducing bacterium Desulfovibrio vulgaris caused severe microbiologically influenced corrosion of zinc and galvanized steel

The microbiologically influenced corrosion (MIC) of zinc and galvanized steel caused by a sulfate reducing bacterium (SRB) was investigated. After 7 days of incubation of Desulfovibrio vulgaris in 125 mL anaerobic vials (100 mL culture medium) at 37 °C, the sessile cell coverage on the galvanized steel was slightly higher than that on pure zinc: (1.9 ± 0.2) × 109 cells/cm2 vs. (9.0 ± 1.8) × 108 cells/cm2. The weight losses for galvanized steel and pure zinc were 31.5 ± 2.5 mg/cm2 and 35.4 ± 4.5 mg/cm2, respectively, which were 101 higher than that for carbon steel. The corrosion current densities of galvanized and pure zinc were 25.5 μA/cm2 and 100 μA/cm2, respectively after the 7-day incubation, confirming that galvanized steel was less prone to SRB MIC despite having a slightly higher sessile cell count. In both cases, the corrosion product was mainly ZnS. Three MIC mechanisms were possible for the severe corrosion against the two metals. Extracellular electron transfer MIC (EET-MIC) was thermodynamically favorable for zinc. Furthermore, in the presence of Zn coupons, H2 evolution in the headspace was 5.5 times higher than without Zn coupons, which suggested that proton attack and/or H2S attack also occurred in the corrosion process.

Characterization and control of biofilms of Salmonella Minnesota of poultry origin

Biofilms characterize sessile form that allows bacterial maintenance under hostile conditions. Salmonella represents an important foodborne zoonotic agent, capable of forming biofilms on diverse surfaces. The emergence of Salmonella Minnesota in the Brazilian poultry production expresses the need for more specific knowledge related to its maintenance in the environment and consequent food contamination, as well as control measures. Our approach combines the analysis of genetic determinants linked to biofilm formation and the phenotypic study of biomass on different surfaces, together with the determination of the effect of chemical agents on the control of sessile structure. It also evaluates the genetic similarity profile of 29 S. Minnesota strains isolated from food and environmental samples in two full-cycle poultry industries from 2009 to 2014. Genetic analysis showed low heterogeneity with the identification of six clonal groups, three clusters with homology greater than 80% and seven distinct genotypes. It was compatible with the presence of important genes in biofilm formation, with 28/29 (97.0%) presenting adrA and csgD and 27 (93.0%) presenting the luxS gene. Allied to this, 19/29 of the strains (66.0%) presented biomass that varied the intensity in weak (9/19–47.0%), moderate (6/19–32.0%) and strong (4/19–21.0%), with characteristic ultrastructure of a mature biofilm. Sodium hypochlorite showed greater efficiency in the control of sessile cells, even after incubation process. The study shows the presence of biofilm-producing S. Minnesota characterizing the problem of persistence in broiler slaughterhouses and indicates as a control strategy the use of sodium hypochlorite in an appropriate manner.

Self-propelling and rolling of a sessile-motile aggregate of the bacterium Caulobacter crescentus

Active dispersal of microorganisms is often attributed to the cells’ motile organelles. However, much less is known about whether sessile cells can access such motility through aggregation with motile counterparts. Here, we show that the rosette aggregates of the bacterium Caulobacter crescentus, although predominantly sessile, can actively disperse through the flagellar motors of motile members. Comparisons in kinematics between the motile rosettes and solitary swimming cells indicate that the rosettes can be powered by as few as a single motor. We further reconstructed the 3D movements of the rosettes to reveal that their proximity to a solid-liquid interface promotes a wheel-like rolling, as powered by the flagellar torque. This rolling movement also features a sequence of sharp turns, a reorientation mechanism distinct from that of swimming cells. Overall, our study elucidates an unexplored regime of aggregation-based motility that can be widely applied to sessile-motile composites.

Exogenous fungal quorum sensing molecules inhibit planktonic cell growth and modulate filamentation and biofilm formation in the Sporothrix schenckii complex

This study investigated the effect of the quorum sensing molecules (QSMs) farnesol, 2-phenylehtanol, tyrosol and tryptophol against planktonic cells, filamentation and biofilms of Sporothrix spp. The antifungal activity of QSMs was evaluated by broth microdilution. QSMs showed MICs in the ranges of 0.01-1 µM (farnesol), 1-8 mM (2-phenylehtanol and tyrosol), and >16 mM (tryptophol). Filamentous biofilm formation was inhibited by farnesol and 2-phenylehtanol and stimulated by tyrosol. Yeast biofilm formation was inhibited by 2-phenylehtanol and tyrosol. Tryptophol did not affect Sporothrix biofilm formation. QSMs showed MICs against mature biofilms of 8-32 µM (farnesol), 8-32 mM (2-phenylehtanol) and 64-128 mM (tyrosol). In conclusion, farnesol, 2-phenylethanol and tyrosol have antifungal activity against planktonic and sessile cells and modulate filamentation and biofilm formation in Sporothrix spp.

A Mixture of D-Amino Acids Enhances the Biocidal Efficacy of CMIT/MIT Against Corrosive Vibrio harveyi Biofilm.

Biocides are widely used for the mitigation of microbial contamination, especially in the field of the aviation fuel industry. However, the long-term use of biocide has raised the concerns regarding the environmental contamination and microbial drug resistance. In this study, the effect of a mixture of D-amino acids (D-tyrosine and D-methionine) on the enhancement of the bactericidal effect of 5-Chloro-2-Methyl-4-isothiazolin-3-one/2-Methyl-2H-isothiazole-3-one (CMIT/MIT) against corrosive Vibrio harveyi biofilm was evaluated. The results revealed that D-Tyr and D-Met alone can enhance the biocidal efficacy of CMIT/MIT, while the treatment of 5 ppm CMIT/MIT, 1 ppm D-Tyr and 100 ppm D-Met showed the best efficacy comparable to that of 25 ppm CMIT/MIT alone. The triple combination treatment successfully prevented the establishment of the corrosive V. harveyi biofilm and effectively removed the mature V. harveyi biofilm. These conclusions were confirmed by the results of sessile cell counts, images obtained by scanning electron microscope and confocal laser scanning microscope, and the ATP test kit.

A Two-Way Road: Antagonistic Interaction Between Dual-Species Biofilms Formed by Candida albicans/Candida parapsilosis and Trichophyton rubrum.

Dermatomycoses include superficial fungal infections of the skin and its appendages. Trichophyton rubrum, Candida albicans, and Candida parapsilosis are some of the most prevalent species that cause dermatomycoses. Several studies show a variable predominance of Candida spp. in relation to dermatophytes, especially in onychomycosis and the possibility of isolating both from the same site. The ability of dermatophytes to form biofilms recently been explored and there is currently no evidence on the involvement of these filamentous fungi in multi-species biofilms. Thus, this study aims to investigate the probable dual-species interaction between T. rubrum and C. albicans and T. rubrum and C. parapsilosis biofilms, considering variable formation conditions, as well as the susceptibility of these dual-species biofilms against terbinafine and efinaconazole. Three conditions of formation of dual-species biofilms were tested: (a) the suspensions of T. rubrum and Candida albicans or C. parapsilosis placed together; (b) suspensions of C. albicans and C. parapsilosis added the pre-adhesion of T. rubrum biofilms; (c) after the maturation of T. rubrum sessile cells. In the first and second conditions, the quantification of metabolic activities, biomass, and polysaccharide materials of mixed biofilms tended to resemble Candida monospecies biofilms. In the third condition, the profiles were modified after the addition of Candida, suggesting that T. rubrum biofilms served as substrate for the development of Candida biofilms. Scanning electron microscopy showed Candida predominance, however, numerous blastoconidia were noted, most evident in the conditions under which Candida was added after the pre-adhesion and maturation of T. rubrum biofilms. Despite the predominance of Candida, the presence of T. rubrum appears to inhibit C. albicans filamentation and C. parapsilosis development, confirming an antagonistic interaction. Fungal burden assays performed when the biofilms were formed together confirmed Candida predominance, as well as susceptibility to antifungals. Further studies will be needed to identify the components of the Candida and T. rubrum biofilm supernatants responsible for inhibiting dermatophyte growth and C. albicans filamentation.

Taking Advantage of Plant Defense Mechanisms to Promote Human Health. The Plant Immune System. First of Two Parts.

Despite the evidence that plants do not possess sessile cells, they are able to mount a vigorous immune response against invaders or under stressful conditions. Plants are endowed with pattern recognition receptors (PPRs) which perceive damage-associated molecular patterns and microbe- associated molecular patterns or pathogen-associated molecular patterns (PAMPs), respectively. PPR activation leads to either the initiation of PAMP-triggered immunity (PTI) (early response) or the effector-triggered immunity (ETI). Both PTI and ETI contribute to plant systemic acquired resistance as an expression of immunological memory or trained immunity. PTI is initiated by activation of both receptor-like kinases and receptor-like proteins, while ETI depends on nucleotide- binding leucine-rich-repeat protein receptors for microbe recognition. Plant chloroplasts contribute to both PTI and ETI through the production of peptides, which act as hormones or phytocytokines. Salicylic acid, jasmonic acid and ethylene are the major compounds involved in plant defense. The interaction between plant receptors and/or their products and bacterial components will be discussed. Also, emphasis will be placed on plant microbiome for its contribution to plant immune response. Finally, the mutual interplay between insects and plants will also be illustrated. A better knowledge of plant immunity may pave the way for the exploitation of plant derivatives in the field of agriculture and medicine, as well.

The Penicillium chrysogenum Q176 Antimicrobial Protein PAFC Effectively Inhibits the Growth of the Opportunistic Human Pathogen Candida albicans.

Small, cysteine-rich and cationic antimicrobial proteins (AMPs) from filamentous ascomycetes promise treatment alternatives to licensed antifungal drugs. In this study, we characterized the Penicillium chrysogenum Q176 antifungal protein C (PAFC), which is phylogenetically distinct to the other two Penicillium antifungal proteins, PAF and PAFB, that are expressed by this biotechnologically important ascomycete. PAFC is secreted into the culture broth and is co-expressed with PAF and PAFB in the exudates of surface cultures. This observation is in line with the suggested role of AMPs in the adaptive response of the host to endogenous and/or environmental stimuli. The in silico structural model predicted five β-strands stabilized by four intramolecular disulfide bonds in PAFC. The functional characterization of recombinant PAFC provided evidence for a promising new molecule in anti-Candida therapy. The thermotolerant PAFC killed planktonic cells and reduced the metabolic activity of sessile cells in pre-established biofilms of two Candida albicans strains, one of which was a fluconazole-resistant clinical isolate showing higher PAFC sensitivity than the fluconazole-sensitive strain. Candidacidal activity was linked to severe cell morphology changes, PAFC internalization, induction of intracellular reactive oxygen species and plasma membrane disintegration. The lack of hemolytic activity further corroborates the potential applicability of PAFC in clinical therapy.

Sodium alginate-based edible coating containing nanoemulsion of Citrus sinensis essential oil eradicates planktonic and sessile cells of food-borne pathogens and increased quality attributes of tomatoes

There is a growing interest from the worldwide scientific community in formulating edible- biodegradable coatings to replace non-biodegradable and expensive commercial wax-based coatings for preserving postharvest quality attributes of vegetables including tomatoes. Postharvest tomatoes are a suspected vehicle for both Salmonella and Listeria in food poisoning incidents. In this work, the effectiveness of edible nano-emulsion coatings containing sweet orange essential oil and sodium alginate were prepared and characterized, then evaluated antibacterial and antibiofilm activity against Salmonella and Listeria and simultaneously, examined its coating effect on various quality characteristics of tomatoes at 22 ± 2 °C over a 15 days storage period. DLS (Dynamic light scattering) study revealed stable nanoemulsion formulation with 43.23 nm particle size. The high whiteness index of nanoemulsion has a positive impact on product marketability and desirability. Antibacterial and antibiofilm studies revealed nanoemulsion effectively eradicate both sessile and planktonic forms of Salmonella and Listeria in both single and multi-species culture conditions. Tomatoes coated with edible coating significantly enhanced firmness up to 33%, decreased total mesophilic bacteria including Salmonella and Listeria, and reduced weight loss up to 3 fold lower than uncoated one. Sensory analysis revealed that the use of the edible coating increased the total acceptance scores of tomatoes.

Study of Microbially Influenced Corrosion in the Presence of Iron-Oxidizing Bacteria (Strain DASEWM2)

The present study deals with the investigation of microbially influenced corrosion of mild steel by the iron-oxidizing bacterium (IOB), Pseudomonas sp. strain DASEWM2, using electrochemical tests, immersion tests, and surface analysis. The corrosion rate obtained by Tafel plots was higher after exposure of ~ 24 h to 72 h when the concentration of sessile cells and constituents of extracellular polymeric substances were high. In inoculated media, cyclic polarization curves show lower pitting potential, repassivation potential, and large area under the hysteresis loop, indicating higher susceptibility of steel to localized corrosion. Immersion tests show higher corrosion rate (0.252 ± 0.05 mpy) and large deep pits in the open area (maximum pit depth 140 ± 8 µm) and under crevice (maximum pit depth 95 ± 4 µm) indicating a higher degree of corrosion in inoculated media. Corrosion products formed on corroded steel samples were analyzed using EDAX, XRD, and FTIR techniques. Goethite and lepidocrocite are the main corrosion products in the case of inoculated media. In contrast, lepidocrocite and iron hydrogen phosphate are the main corrosion products in the case of control media. The absence of iron hydrogen phosphate, a protective type of corrosion product, on steel exposed to inoculated media may be responsible for observing a higher degree of corrosion due to bacteria.

Investigation of Biocidal Effect of Microfiltration Membranes Impregnated with Silver Nanoparticles by Sputtering Technique.

Silver nanoparticles were loaded in microfiltration membranes by sputtering technique for the development of biocidal properties and biofouling resistance. This technology allows good adhesion between silver nanoparticles and the membranes, and fast deposition rate. The microfiltration membranes (15 wt.% polyethersulfone and 7.5 wt.% polyvinylpyrrolidone in N,N-dimethylacetamide) were prepared by phase inversion method, and silver nanoparticles were deposited on their surface by the physical technique of vapor deposition in a sputtering chamber. The membranes were characterized by Field Emission Scanning Electron Microscopy, and the presence of silver was investigated by Energy-Dispersive Spectroscopy and X-ray Diffraction. Experiments of silver leaching were carried out through immersion and filtration tests. After 10 months of immersion in water, the membranes still presented ~90% of the initial silver, which confirms the efficiency of the sputtering technique. Moreover, convective experiments indicated that 98.8% of silver remained in the membrane after 24 h of operation. Biocidal analyses (disc diffusion method and biofouling resistance) were performed against Pseudomonas aeruginosa and confirmed the antibacterial activity of these membranes with 0.6 and 0.7 log reduction of viable planktonic and sessile cells, respectively. These results indicate the great potential of these new membranes to reduce biofouling effects.

Proteomic analysis reveals the temperature-dependent presence of extracytoplasmic peptidases in the biofilm exoproteome of Listeria monocytogenes EGD-e.

The foodborne pathogen Listeria monocytogenes resists environmental stresses by forming biofilms. Because this pathogen transmits between the environment and the host, it must adapt to temperature as an environmental stress. In this study, we aimed to identify which proteins were present depending on the temperature in the biofilms of L. monocytogenes EGD-e. Proteins in the supernatants of biofilms formed at 25°C and 37°C were compared using two-dimensional gel electrophoresis and liquid chromatography with tandem mass spectrometry. The larger number of extracytoplasmic proteins associated with cell wall/membrane/envelop biogenesis was identified from the supernatant of biofilms formed at 25°C (7) than those at 37°C (0). Among the 16 extracytoplasmic proteins detected only at 25°C, three were peptidases, namely Spl, Cwh, and Lmo0186. Moreover, mRNA expression of the three peptidases was higher at 25°C than at 37°C. Interestingly, this adaptation of gene expression to temperature was present in sessile cells but not in dispersed cells. After inhibiting the activity of extracytoplasmic peptidases with a protease inhibitor, we noted that the levels of biofilm biomass increased with higher concentrations of the protease inhibitor only when L. monocytogenes grew biofilms at 25°C and not at 37°C. Overall, our data suggest an effect of temperature on the presence of peptidases in L. monocytogenes biofilms. Additionally, increasing the levels of extracytoplasmic peptidases in biofilms is likely a unique feature for sessile L. monocytogenes that causes a naturally occurring breakdown of biofilms and facilitates the pathogen exiting biofilms and disseminating into the environment.