Sessile Cells Research Articles

Determining conditions for inactivation of multispecies biofilm cells by peracetic acid applying response surface methodology

Biofilms can form on various surfaces and are typically associated with multiple microbial species. Particularly in the food industry, biofilms are often a source of contamination, necessitating mitigation strategies. In this study, Response Surface Methodology (RSM) was utilized to optimize conditions for the inactivation of multispecies biofilm cells, with peracetic acid (PAA) (0.05% to 0.5%), treatment time (5 to 30 minutes), and temperature (25 to 60 ºC) as independent variables. The multispecies biofilm consisted of Gram-positive and Gram-negative bacteria previously isolated from biofilms formed in the presence of raw milk. Stainless steel (SS) coupons were immersed in reconstituted whole milk inoculated with Pseudomonas fluorescens, Rahnella inusitata, Staphylococcus aureus, and Micrococcus aloeverae. Sessile cell counts of approximately 108 CFU/cm2 were obtained after 10 days of incubation at 4 ºC. The response surface model exhibited a good fit, with R2 of 0.949 and adjR2 of 0.883. All independent variables had a positive effect on the inactivation of biofilm cells. The maximum inactivation achieved was approximately 7 log (CFU/cm2), with the highest values observed at 0.5% PAA, for 30 min at 60°C. Epifluorescence microscopy revealed that many cells in the biofilm were dead or injured after treatment at the central point (PAA = 0.275%; Time = 17.5 min; Temperature = 42.5 ºC). RSM helps to predict better conditions for maximum biofilm eradication and proves to be a promising approach for monitoring the inactivation of multispecies biofilm cells, which warrants further exploration.

Time-dependent corrosion behavior of EH36 steel caused by Pseudomonas aeruginosa based on big data monitoring technology

Marine microbial corrosion poses a significant threat to the safe service of marine engineering equipment. Previous studies have often failed to thoroughly analyze the continuous and prolonged microbial corrosion process, resulting in an incomplete understanding of microbial corrosion mechanisms involved at various stages and the development of ineffective control strategies. This study employed a corrosion big data online real-time monitoring technique to investigate the time-dependent corrosion behavior of EH36 steel caused by Pseudomonas aeruginosa in aerobic environments over a 30-d incubation period. It was found that P. aeruginosa accelerated the corrosion of EH36 steel in the early stages by enhancing the cathodic oxygen reduction process. As oxygen levels declined, P. aeruginosa transitioned from aerobic to anaerobic respiration, promoting corrosion through biocatalytic nitrate reduction. In the later stages, the reduction in sessile cell counts, extreme low oxygen concentration, and dense surface film increased the charge transfer and film resistances, ultimately leading to corrosion inhibition. The weight loss and electrochemical data confirmed the effectiveness of the big data monitoring technique in investigating microbial corrosion, which provides new approaches for diagnosing and preventing microbial corrosion.

Inhibitory effect of Bacillus velezensis 1273 strain cell-free supernatant against developing and preformed biofilms of Staphylococcus aureus and MRSA

Microbial biofilms constitute a significant virulence factor and a substantial challenge in clinical environments due to their role in promoting antimicrobial resistance and their resilience to eradication efforts. Methicillin-resistant Staphylococcus aureus (MRSA) infections substantially increase healthcare costs, extend hospitalizations, and elevate morbidity and mortality rates. Therefore, developing innovative strategies to target and eliminate these bacteria and their biofilms effectively is imperative for robust epidemiological control. In this study, we evaluated the antibacterial and antibiofilm activities of cell-free supernatant (CFS) obtained from the Bacillus velezensis 1273 strain culture. Our data showed that CFS inhibited the growth of S. aureus ATCC 29213 and MRSA (clinical strain), with greater efficacy observed against S. aureus (1:16 dilution). Furthermore, CFS showed substantial potential in reducing biofilm formation in both strains (∼30 %) at subinhibitory concentrations. Additionally, the antibacterial activity against biofilm-formed cells showed that pure CFS treatment decreased the viability of S. aureus (60 %) and MRSA (45 %) sessile cells. We further demonstrated that CFS treatment induces the production of reactive oxygen species (ROS) and damages the membranes and cell walls of the pathogen cells. Genome analysis revealed the presence of genes encoding bacteriocins and secondary metabolites with antibacterial activity in the B. velezensis 1273 genome. These findings highlight the potential of probiotic bacterial metabolites as antibiofilm and anti-multidrug-resistant pathogens.

Exploring NRB Biofilm Adhesion and Biocorrosion in Oil/Water Recovery Operations Within Pipelines.

Microbially influenced corrosion represents a critical challenge to the integrity and durability of carbon steel infrastructure, particularly in environments conducive to biofilm formation by nitrate-reducing bacteria (NRB). This study investigated the impact of NRB biofilms on biocorrosion processes within oil/water recovery operations in Algerian pipelines. A comprehensive suite of experimental and analytical techniques, including microbial analysis, gravimetric methods, and surface characterization, were employed to elucidate the mechanisms of microbially influenced corrosion (MIC). Weight loss measurements revealed that carbon steel samples exposed to injection water exhibited a corrosion rate of 0.0125 mm/year, significantly higher than the 0.0042 mm/year observed in crude oil environments. The microbial analysis demonstrated that injection water harbored an average of (4.4 ± 0.56) × 106 cells/cm2 for sessile cells and (3.1 ± 0.25) × 105 CFU/mL for planktonic cells, in stark contrast to crude oil, which contained only (2.4 ± 0.34) × 103 cells/cm2 for sessile cells and (4.5 ± 0.12) × 102 CFU/mL for planktonic cells, thereby highlighting the predominant role of injection water in facilitating biofilm formation. Contact angle measurements of injection water on carbon showed 45° ± 2°, compared to 85° ± 4° for crude oil, suggesting an increased hydrophilicity associated with enhanced biofilm adhesion. Scanning electron microscopy further confirmed the presence of thick biofilm clusters and corrosion pits on carbon steel exposed to injection water, while minimal biofilm and corrosion were observed in the crude oil samples.

Investigating the antimicrobial and antibiofilm properties of marine halophilic Bacillus species against ESKAPE pathogens.

Antimicrobial resistance (AMR), known as the "silent pandemic," is exacerbated by pathogenic bacteria's ability to form biofilms. Marine compounds hold promise for novel antibacterial drug discovery. Two isolates from preliminary saltwater environment screening demonstrated antimicrobial activity and were subsequently identified as Bacillus subtilis MTUA2 and Bacillus velezensis MTUC2. Minimum inhibitory concentrations (MICs), minimum biofilm inhibition concentrations (MBICs) and minimum biofilm eradication concentrations (MBECs) required to prevent and/or disrupt bacterial growth and biofilm formation were established for MRSA, Staphylococcus aureus, Acinetobacter baumannii and Escherichia coli. The metabolic activity within biofilms was determined by the 2,3,5-triphenyltetrazolium chloride assay. Both Bacillus species exhibited unique antimicrobial effects, reducing MRSA and S. aureus planktonic cell growth by 50% and sessile cell growth for S. aureus and E. coli by 50% and 90%, respectively. No effect was observed against A. baumannii. Significant MBIC and MBEC values were achieved, with 99% inhibition and 90% reduction in MRSA and S. aureus biofilms. Additionally, 90% and 50% inhibition was observed in E. coli and A. baumannii biofilms, respectively, with a 50% reduction in E. coli biofilm. These findings suggest that the mode of action employed by B. subtilis MTUA2 and B. velezensis MTUC2 metabolites should be further characterized and could be beneficial if used independently or in combination with other treatments.

Fluid depth-dependent biofilm characteristics and their influence on sulfate-reducing bacteria-induced corrosion of carbon steel

This study investigates the response of biofilm characteristics to variations in fluid depth and their influence on the corrosion behavior of carbon steel (C1020) under low-flow fluid conditions, utilizing Desulfovibrio vulgaris. The experiments were conducted in an anaerobic chamber at 30 °C, utilizing modified Baar's medium as the testing medium. The findings reveal that fluid depth significantly impacts biofilm-corrosion product composite formation, with deeper depths promoting thicker and more heterogeneous biofilm-corrosion product layer compared to shallower depths, where a thinner and more uniform biofilm-corrosion product layer is observed. Moreover, the characteristics of initially attached biofilms was verified as the primary factor affecting subsequent corrosion behavior during prolonged exposure. Corrosion analysis reveals that greater fluid depth leads to increased weight loss (91 ± 13.2 mg/cm2) and deeper pit depths (540 ± 69 μm), surpassing those observed in shallower test media (21 ± 2.3 mg/cm2 and 105 ± 17 μm) after 28 days of exposure. The corrosion products within the biofilm were predominantly FeS and Fe3(PO4)2·8H2O. A direct relationship was observed between the thickness of this biofilm-corrosion product layer and the progression of pit depth, suggesting a strong correlation between carbon steel corrosion and biofilm development in limited fluid depths (e.g., 5–15 mm). Furthermore, a significant association between the deepest pits (average) and the number of sessile cells within the biofilm underscores the pivotal role of sessile cell numbers in carbon steel corrosion.

Effect of exposure to sub-inhibitory concentrations of biocides on the susceptibility to antibiotics of sessile cells of Listeria monocytogenes and Salmonellaenterica

The effect of sub-inhibitory concentrations of several disinfectants on the antibiotic resistance of sessile cells of Listeria monocytogenes and Salmonella enterica was investigated. Biofilms were formed in the presence of biocides at MIC/2. Five compounds were used: peracetic acid (PAA), benzalkonium chloride (BZK), sodium hypochlorite (SHY), polyvinyl pyrrolidone iodine (PVI) and bis(3-aminopropyl)amine (BAPA). Two strains of L. monocytogenes (LM30 and LM28) and S. enterica (S. Typhimurium -ST- and S. Enteritidis -SE-) were used. Mono-species biofilms (MSB) and mixed biofilms (MB, LM30 + ST and LM28 + SE) were tested. LM30 went from susceptible to resistant to tetracycline and erythromycin after exposure to BAPA in MB. LM28 went from susceptible to resistant to tetracycline in the presence of PVI in MSB, from susceptible to intermediate to chloramphenicol after exposure to PAA in both MSB and MB, from susceptible to resistant to erythromycin after exposure to BAPA in MB, and from susceptible to resistant to ampicillin after exposure to SHY and BAPA in MB. ST went from intermediate to resistant to gentamicin after exposure to SHY, PVI and BAPA (MSB), or to PVI and BAPA (MB). This strain went from susceptible to intermediate to cefoxitin after exposure to BZK and PVI (MSB) or to BZK, PVI and BAPA (MB). For fosfomycin, ST went from susceptible to intermediate after exposure to PAA, PVI and BAPA (MSB) or to SHY, BZK, PVI and BAPA (MB). In the case of cephalothin, ST moved from intermediate to resistant category in MSB exposed to BAPA. SE moved from susceptible to intermediate to cefoxitin after contact with PAA in both MSB and MB. By contrast, in several occasions, strains increased their susceptibility to antibiotics after exposure to biocides. In some additional cases, contact with disinfectants caused significative modifications in the MIC of various antibiotics without the strains changing from one category to another. These findings show that contact with low concentrations of biocides can modify the antibiotic resistance of sessile cells of L. monocytogenes and S. enterica. The need to use adequate doses of disinfectants in food facilities in order to avoid changes in bacterial resistance is highlighted.

Fluopsin C Promotes Biofilm Removal of XDR Acinetobacter baumannii and Presents an Additive Effect with Polymyxin B on Planktonic Cells.

Acinetobacter baumannii emerged as one of the most important pathogens for the development of new antimicrobials due to the worldwide detection of isolates resistant to all commercial antibiotics, especially in nosocomial infections. Biofilm formation enhances A. baumannii survival by impairing antimicrobial action, being an important target for new antimicrobials. Fluopsin C (FlpC) is an organocupric secondary metabolite with broad-spectrum antimicrobial activity. This study aimed to evaluate the antibiofilm activity of FlpC in established biofilms of extensively drug-resistant A. baumannii (XDRAb) and the effects of its combination with polymyxin B (PolB) on planktonic cells. XDRAb susceptibility profiles were determined by Vitek 2 Compact, disk diffusion, and broth microdilution. FlpC and PolB interaction was assessed using the microdilution checkerboard method and time-kill kinetics. Biofilms of XDRAb characterization and removal by FlpC exposure were assessed by biomass staining with crystal violet. Confocal Laser Scanning Microscopy was used to determine the temporal removal of the biofilms using DAPI, and cell viability using live/dead staining. The minimum inhibitory concentration (MIC) of FlpC on XDRAb was 3.5 µg mL-1. Combining FlpC + PolB culminated in an additive effect, increasing bacterial susceptibility to both antibiotics. FlpC-treated 24 h biofilms reached a major biomass removal of 92.40 ± 3.38% (isolate 230) using 7.0 µg mL-1 FlpC. Biomass removal occurred significantly over time through the dispersion of the extracellular matrix and decreasing cell number and viability. This is the first report of FlpC's activity on XDRAb and the compound showed a promissory response on planktonic and sessile cells, making it a candidate for the development of a new antimicrobial product.

Exploring the Antibiofilm Effect of Sertraline in Synergy with Cinnamomum verum Essential Oil to Counteract Candida Species.

The emergence and spread of drug-resistant pathogens, resulting in antimicrobial resistance, continue to compromise our capability to handle commonly occurring infectious diseases. The rapid global spread of multi-drug-resistant pathogens, particularly systemic fungal infections, presents a significant concern, as existing antimicrobial drugs are becoming ineffective against them. In recent decades, there has been a notable increase in systemic fungal infections, primarily caused by Candida species, which are progressively developing resistance to azoles. Moreover, Candida species biofilms are among the most common in clinical settings. In particular, they adhere to biomedical devices, growing as a resilient biofilm capable of withstanding extraordinarily high antifungal concentrations. In recent years, many research programs have concentrated on the development of novel compounds with possible antimicrobial effects to address this issue, and new sources, such as plant-derived antimicrobial compounds, have been thoroughly investigated. Essential oils (EOs), among their numerous pharmacological properties, exhibit antifungal, antibacterial, and antiviral activities and have been examined at a global scale as the possible origin of novel antimicrobial compounds. A recent work carried out by our research group concerned the synergistic antibacterial activities of commercially available and chemically characterized Cinnamomum verum L. essential oil (C. verum EO) in association with sertraline, a selective serotonin reuptake inhibitor whose repositioning as a non-antibiotic drug has been explored over the years with encouraging results. The aim of this work was to explore the synergistic effects of C. verum EO with sertraline on both planktonic and sessile Candida species cells. Susceptibility testing and testing of the synergism of sertraline and C. verum EO against planktonic and sessile cells were performed using a broth microdilution assay and checkerboard methods. A synergistic effect was evident in both the planktonic cells and mature biofilms, with significant reductions in fungal viability. Indeed, the fractional inhibitory concentration index (FICI) was lower than 0.5 for all the associations, thus indicating significant synergism of the associations with the Candida strains examined. Moreover, the concentrations of sertraline able to inhibit Candida spp. strain growth and biofilm formation significantly decreased when it was used in combination with C. verum EO for all the strains considered, with a reduction percentage in the amount of each associated component ranging from 87.5% to 97%.

Antifungal effect of a metabolite of Pseudomonas aeruginosa LV strain on azole-resistant Candida albicans

Candida albicans remains the most common agent of candidiasis worldwide. This yeast is generally sensitive to most antifungals, however, the emergence of azole-resistant C. albicans has been reported. In addition, this microorganism can form biofilms on various surfaces, making it difficult to treat infections. In this study, the effect of secondary metabolites of Pseudomonas aeruginosa strain LV on planktonic and sessile cells of C. albicans, with different genotypes and susceptibility profile to fluconazole and voriconazole, was evaluated. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the semi-purified fraction F4a ranged from 1.56 to 6.25 μg/mL and 6.25 to 25 μg/mL, respectively. Fluopsin C appears to be the antifungal component of F4a. The semi-purified fraction and fluopsin C showed fungicidal activity, dose and time dependent. F4a caused severe damage to the morphology and ultrastructure of planktonic fungal cells, and significantly reduced the viability of 24-hour biofilms, with MIC for sessile cells from 12.5 to 25.0 μg/mL. However, cytotoxicity was detected in mammalian cells for F4a and fluopsin C at concentrations that showed antifungal activity. These results indicate that fluopsin C may be a prototype for the development of new antifungals for C. albicans.

Geraniol Potentiates the Effect of Fluconazole against Planktonic and Sessile Cells of Azole-Resistant Candida tropicalis: In Vitro and In Vivo Analyses.

Candida tropicalis is regarded as an opportunistic pathogen, causing diseases ranging from superficial infections to life-threatening disseminated infections. The ability of this yeast to form biofilms and develop resistance to antifungals represents a significant therapeutic challenge. Herein, the effect of geraniol (GER), alone and combined with fluconazole (FLZ), was evaluated in the planktonic and sessile cells of azole-resistant C. tropicalis. GER showed a time-dependent fungicidal effect on the planktonic cells, impairing the cell membrane integrity. Additionally, GER inhibited the rhodamine 6G efflux, and the molecular docking analyzes supported the binding affinity of GER to the C. tropicalis Cdr1 protein. GER exhibited a synergism with FLZ against the planktonic and sessile cells, inhibiting the adhesion of the yeast cells and the viability of the 48-h biofilms formed on abiotic surfaces. C. tropicalis biofilms treated with GER, alone or combined with FLZ, displayed morphological and ultrastructural alterations, including a decrease in the stacking layers and the presence of wilted cells. Moreover, neither GER alone nor combined with FLZ caused toxicity, and both treatments prolonged the survival of the Galleria mellonella larvae infected with azole-resistant C. tropicalis. These findings indicate that the combination of GER and FLZ may be a promising strategy to control azole-resistant C. tropicalis infections.

A New Convenient Method to Assess Antibiotic Resistance and Antimicrobial Efficacy against Pathogenic Clostridioides difficile Biofilms.

Clostridioides difficile is a widely distributed anaerobic pathogen. C. difficile infection is a serious problem in healthcare. Its biofilms have been found to exhibit biocorrosivity, albeit very little, but sufficient for it to correlate with biofilm growth/health. This work demonstrated the use of a disposable electrochemical biofilm test kit using two solid-state electrodes (a 304 stainless steel working electrode, and a graphite counter electrode, which also served as the reference electrode) in a 10 mL serum vial. It was found that the C. difficile 630∆erm Adp-4 mutant had a minimum inhibitory concentration (MIC) for vancomycin twice that of the 630∆erm wild type strain in biofilm prevention (2 ppm vs. 1 ppm by mass) on 304 stainless steel. Glutaraldehyde, a commonly used hospital disinfectant, was found ineffective at 2% (w/w) for the prevention of C. difficile 630∆erm wild type biofilm formation, while tetrakis(hydroxymethyl)phosphonium sulfate (THPS) disinfectant was very effective at 100 ppm for both biofilm prevention and biofilm killing. These antimicrobial efficacy data were consistent with sessile cell count and biofilm imaging results. Furthermore, the test kit provided additional transient biocide treatment information. It showed that vancomycin killed C. difficile 630∆erm wild type biofilms in 2 d, while THPS only required minutes.

Preliminary investigations of microbiologically influenced corrosion of 304 stainless steel by anaerobic Clostridioides difficile biofilm

Clostridioides difficile is a pathogenic anaerobe that potentially causes microbiologically influenced corrosion (MIC). Coupons of 304 stainless steel (SS) were incubated with C. difficile in deoxygenated brain heart infusion supplement medium. After a 7-d incubation, C. difficile biofilms were observed on the 304 SS coupon surfaces. The sessile cell count on 304 SS coupons were (1.9 ± 0.5) × 107 cells/cm2. It was found that this high-grade SS did not suffer measurable corrosion weight loss and pitting. X65 carbon steel was used to verify C. difficile bio-corrosivity. A 7-d weight loss of 0.9 ± 0.2 mg/cm2 was found on X65 coupons with the same incubation condition, which manifested as uniform corrosion. 13%Cr steel, also known as 420 SS which is a low-grade SS that is prone to pitting, was used to verify pitting by C. difficile. A 15.2 μm pit was observed after 26 d of incubation. Electrochemical tests were conducted in a 10 mL biofilm/MIC test kit. The electrochemical analysis of electron mediator injection indicated that MIC of 304 SS by C. difficile belongs to extracellular electron transfer-MIC. A 100 ppm (w/w) tetrakis (hydroxymethyl)phosphonium sulfate (a green biocide) injection test proved that it is a suitable disinfectant for C. difficile.

Active interface bulging in Bacillus subtilis swarms promotes self-assembly and biofilm formation

Microbial communities such as biofilms are commonly found at interfaces. However, it is unclear how the physical environment of interfaces may contribute to the development and behavior of surface-associated microbial communities. Combining multimode imaging, single-cell tracking, and numerical simulations, here, we found that activity-induced interface bulging promotes colony biofilm formation in Bacillus subtilis swarms presumably via segregation and enrichment of sessile cells in the bulging area. Specifically, the diffusivity of passive particles is ~50% lower inside the bulging area than elsewhere, which enables a diffusion-trapping mechanism for self-assembly and may account for the enrichment of sessile cells. We also uncovered a quasilinear relation between cell speed and surface-packing density that underlies the process of active interface bulging. Guided by the speed-density relation, we demonstrated reversible formation of liquid bulges by manipulating the speed and local density of cells with light. Over the course of development, the active bulges turned into striped biofilm structures, which eventually give rise to a large-scale ridge pattern. Our findings reveal a unique physical mechanism of biofilm formation at air-solid interface, which is pertinent to engineering living materials and directed self-assembly in active fluids.

The combined effect of pH and NaCl on the susceptibility of Listeria monocytogenes to rhamnolipids

The use of natural and sustainable additives, that are less aggressive to the environment, is a trend in the food industry. Rhamnolipids (RL) biosurfactants have shown potential for controlling food pathogens however, due to the presence of free carboxyl groups, the pH and ionic strength may influence the properties of such surfactants. In this study, we describe the antimicrobial activity of RL under different pH values and NaCl concentrations, towards both planktonic and biofilms of Listeria monocytogenes. RL were effective at pH 5.0 and the addition of 5 % NaCl improved the bactericidal efficacy for planktonic and sessile cells. The effect of NaCl was more pronounced at pH above 6 showing a significant increase in RL antimicrobial activity. At pH 7.0 planktonic population was eradicated by RL only when salt was present whereas biofilm viability was decreased by 5 log with MBIC varying from > 2500.0 mg/L (RL) to 39.0 mg/L (RL + 5 % NaCl). Larger vesicular and lamellar RL self-assembly structures were predominant when NaCl was present, suggesting their association with the antimicrobial activity observed. The pH and ionic strength of the medium are important parameters to be considered for the development of RL-based strategies to control L. monocytogenes.

Improvement of a low-cost protocol for a simultaneous comparative evaluation of hydrolytic activity between sessile and planktonic cells: Candida albicans as a study model

Candida albicans is often implicated in nosocomial infections with fatal consequences. Its virulence is contributed to hydrolytic enzymes and biofilm formation. Previous research focused on studying these virulence factors individually. Therefore, this study aimed to investigate the impact of biofilm formation on the hydrolytic activity using an adapted low-cost method. Eleven strains of C. albicans were used. The biofilms were formed on pre-treated silicone discs using 24-well plates and then deposited on the appropriate agar to test each enzyme, while the planktonic cells were conventionally seeded. Biofilms were analysed using Raman spectroscopy, fluorescent and scanning electron microscopy. The adapted method provided an evaluation of hydrolytic enzymes activity in C. albicans biofilm and showed that sessile cells had a higher phospholipase and proteinase activities compared with planktonic cells. These findings were supported by spectroscopic and microscopic analyses, which provided valuable insights into the virulence mechanisms of C. albicans during biofilm formation.

Fe0-to-microbe electron transfer corrosion of EH36 steel by planktonic cells of Desulfovibrio vulgaris mediated by riboflavin

This study investigated the Fe0-to-microbe extracellular electron transfer (EET) corrosion of EH36 steel by planktonic Desulfovibrio vulgaris cells using Tetrakis hydroxymethyl phosphonium sulfate (THPS), peptide E6, and riboflavin (RF) as tools. 60 min after 20 ppm THPS & E6 injection and 50 ppm THPS injection, there was no difference in sessile cell counts, but planktonic cell counts were different. The subsequent injection of 20 ppm RF accelerated D. vulgaris MIC in varying degrees within minutes. This was attributed to RF-mediated EET of planktonic cells to accelerate corrosion, which can be further confirmed by the elevated expression levels of cytochrome c.

Localized corrosion and brittle fracture of X80 carbon steel under tensile stress induced by sulfate reducing bacteria

The microbiologically influenced corrosion (MIC) of X80 carbon steel under varying tensile stresses with the presence of sulfate reducing bacteria (SRB) was investigated. The stress loads affected the SRB sessile cell counts on the carbon steel, and exerted a significant influence on the rate of MIC to which the carbon steel was subjected. Stresses below 400 MPa elicited a modest promotion effect, whereas the stress of 550 MPa which was approaching the yield strength markedly expedited the corrosion of SRB on X80 carbon steel. Finite element simulation revealed that the elastic stress of 550 MPa led to plastic deformation in the stress concentration zone at the base of the corrosion pit, exacerbating the dissolution of the metal in pits and inducing secondary pitting corrosion. Additionally, this work confirmed that SRB promoted the permeation of hydrogen into the carbon steel matrix. The coexistence of high elastic stress and SRB rendered X80 carbon steel more prone to brittle fracture.

Improving the Antimicrobial Potency of Berberine for Endodontic Canal Irrigation Using Polymeric Nanoparticles.

To address the challenges posed by biofilm presence and achieve a substantial reduction in bacterial load within root canals during endodontic treatment, various irrigants, including nanoparticle suspensions, have been recommended. Berberine (BBR), a natural alkaloid derived from various plants, has demonstrated potential applications in dentistry treatments due to its prominent antimicrobial, anti-inflammatory, and antioxidant properties. This study aimed to produce and characterize a novel polymeric nanoparticle of poly (lactic-co-glycolic acid) (PLGA) loaded with berberine and evaluate its antimicrobial activity against relevant endodontic pathogens, Enterococcus faecalis, and Candida albicans. Additionally, its cytocompatibility using gingival fibroblasts was assessed. The polymeric nanoparticle was prepared by the nanoprecipitation method. Physicochemical characterization revealed spheric nanoparticles around 140 nm with ca, -6 mV of surface charge, which was unaffected by the presence of BBR. The alkaloid was successfully incorporated at an encapsulation efficiency of 77% and the designed nanoparticles were stable upon 20 weeks of storage at 4 °C and 25 °C. Free BBR reduced planktonic growth at ≥125 μg/mL. Upon incorporation into PLGA nanoparticles, 20 μg/mL of [BBR]-loaded nanoparticles lead to a significant reduction, after 1 h of contact, of both planktonic bacteria and yeast. Sessile cells within biofilms were also considered. At 30 and 40 μg/mL, [BBR]-loaded PLGA nanoparticles reduced the viability of the sessile endodontic bacteria, upon 24 h of exposure. The cytotoxicity of BBR-loaded nanoparticles to oral fibroblasts was negligible. The novel berberine-loaded polymeric nanoparticles hold potential as a promising supplementary approach in the treatment of endodontic infections.

Comparative analysis of commercial cleaning and disinfection formulations and protocols for effective eradication of biofilms formed by a Pseudomonas fluorescens strain isolated from a poultry meat plant

Pseudomonas spp. is the main genus of spoilage bacteria for meat stored under aerobic chilling conditions. Given their ability to adapt to various environmental conditions and to form biofilms, they are common among residential microbiota in the food industry. Biofilms are complex, structured microbial communities encased in a self-produced exopolysaccharide matrix. Resident biofilm bacteria show increased resistance against disinfectants and dynamic or hostile environments, and are therefore a persistent source of food contamination. Hence, there is a need to design and implement control strategies that are effective against biofilms in the food industry. In this study, we tested a series of commercial cleaning and disinfection agents applied by immersion or aerosolization against robust biofilms of a Pseudomonas fluorescens strain isolated from a poultry meat plant by comparing the sessile cell counts before and after the procedure. The single application of a) an alkaline or an enzymatic detergent, b) a disinfectant based on peracetic acid and hydrogen peroxide, or c) a detergent-disinfectant based on tertiary alkylamines or sodium hypochlorite, failed to completely eradicate the biofilm. However, two-step cleaning and disinfection procedures involving the application of a detergent (either alkaline or enzymatic at 50 °C) followed by the application of the disinfectant based on peracetic acid and hydrogen peroxide at the manufacturer's recommended concentrations did result in the complete eradication (p ≤ 0.05) of the robust biofilm. In addition, cleaning and disinfection procedures applied by aerosolization showed a notable efficacy (p ≤ 0.05) against surface-dried cells and sessile cells of P. fluorescens. Therefore, this approach could be regarded as a potentially viable alternative to commonly applied immersion techniques.