Biofilm Reduction Research Articles

Roles of Response Regulators in the Two-Component System in the Formation of Stress Tolerance, Motility and Biofilm in Salmonella Enteritidis.

Two-component systems (TCS) of Salmonella enterica serovar Enteritidis are composed of a histidine kinase and a response regulator (RR) and represent a critical mechanism by which bacteria develop resistance to environmental stress. Here, we characterized the functions of RRs in TCS in the formation of stress tolerance, motility and biofilm using twenty-six S. Enteritidis RR-encoding gene deletion mutants. The viability results unraveled their essential roles in resistance to elevated temperature (GlrR), pH alterations (GlrR, TctD, YedW, ArcA and YehT), high salt (PhoB, BaeR, CpxR, PhoP, UvrY and TctD), oxidative stress (PhoB, YedW, BaeR, ArcA, PhoP, UvrY, PgtA and QseB) and motility (ArcA, GlnG, PgtA, PhoB, UhpA, OmpR, UvrY and QseB) of S. Enteritidis. The results of the crystal violet staining, microscopy observation and Congo red binding assays demonstrated that the absence of ArcA, GlnG, PhoP, OmpR, ZraR or SsrB in S. Enteritidis led to a reduction in biofilms and an impairment in red/dry/rough macrocolony formation, whereas the absence of UvrY exhibited an increase in biofilms and formed a brown/smooth/sticky macrocolony. The results indicated the regulatory effects of these RRs on the production of biofilm matrix, curli fimbriae and cellulose. Our findings yielded insights into the role of TCSs, making them a promising target for combating S. Enteritidis.

Cryptocarya moschata fractions decrease planktonic cells and biofilms of Candida albicans and Streptococcus mutans.

Extracts of Cryptocarya species have been shown to reduce biofilms, demonstrating their antimicrobial effects. The extracts can be fractionated to optimize their potential. In this study, we evaluated the antimicrobial activity of Cryptocarya moschata fractions against planktonic cells and biofilms of Candida albicans and Streptococcus mutans. Four fractions were prepared: 100% hexane, acetate/hexane 1:1, 100% ethyl acetate, and water. The effect of the fractions on planktonic cells was assessed by counting the colony-forming units per milliliter (CFU/mL). Biofilm tests included CFU/mL, cell metabolic activity, and qualitative analysis using confocal laser scanning microscopy (CLSM). Results were analyzed by the Mann-Whitney U test (α = 0.05). The fractions contained lipophilic constituents, styrylpyrones, glycosylated flavonoids, and alkaloids. Acetate/hexane (1:1) and 100% ethyl acetate fractions reduced the CFU/mL of planktonic C. albicans. C. moschata fractions did not affect planktonic S. mutans. For biofilms, the fractions reduced the CFU/mL (from 2-5 logs) and cell metabolic activity (approximately 80% reduction in a single-species biofilm). CLSM showed the fractions reduced microorganism viability and damaged the extracellular matrix of biofilms. We conclude that the acetate/hexane 1:1 and 100% ethyl acetate C. moschata fractions exhibit antimicrobial effects against biofilms.

Effectiveness of Activated Sodium Hypochlorite Irrigation by Shock Wave-Enhanced Emission Photoacoustic Streaming, Sonic and Ultrasonic Devices in Removing Enterococcus faecalis Biofilm From Root Canal System.

Aim: To compare shock wave-enhanced emission photoacoustic streaming (SWEEPS) with sonic- and ultrasonically activated irrigation systems in removing Enterococcus faecalis biofilm from the root canal system. Methodology: Fifty human single-canalled mandibular premolars were included in the study. After access cavity preparation, the root canals were prepared to a standardized size and taper. Then, the entire root surface was covered with two layers of resin, and the root apices were sealed before sterilization. All root canals were inoculated with E. faecalis biofilm, and the samples were incubated aerobically for 2 weeks at 37 °C. Biofilm formation was confirmed by scanning electron microscopy. All samples were randomly divided into five groups (n = 10 each) based on their irrigation activation method as A (no treatment or negative control), B (no irrigation or positive control), C (sonically activated irrigation (SAI)), D (ultrasonically activated irrigation (UAI)), and E (needle irrigation activated by an Er: YAG laser device using a SWEEPS quartz tip (SWEEPS)). Then, dentine chips were retrieved, vortexed, and diluted for colony-forming unit counts. Data were analysed using analysis of variance and post-hoc Tukey tests (α = 5%). Results: All methods could significantly reduce E. faecalis biofilm compared with control so that the UAI, SWEEPS, and SAI groups indicated a 23.54%, 14.89%, and 7.81% biofilm reduction, respectively. UAI demonstrated a significantly more effective reduction of E. faecalis biofilm than SAI (p = 0.004). Conclusions: All irrigation activation methods significantly reduced E. faecalis biofilm, with ultrasonic use being the most effective.

Phytochemical Analysis and Antioxidant, Antimicrobial, and Antibiofilm Effects of a New Himalayan Lichen Placidium deosaiense Usman and Khalid Growing in Pakistan

Phytochemical composition and antimicrobial, antibiofilm, and antioxidant effects of a newly described Himalayan lichen Placidium deosaiense Usman and Khalid growing in Pakistan were investigated. HPLC–DAD methods were used for identification of secondary metabolites in acetone and methanol extracts. The total phenolics content was measured using a spectrophotometric method. The study investigated the antioxidant (DPPH-scavenging activity assay and reducing-power assay), antibacterial (minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC)), and antibiofilm (inhibition of biofilm formation and reduction in mature biofilm) activities of extracts of the lichen P. deosaiense and isolated parietin. The chemical constituents olivetol, olivetolic acid, haematommic acid, fallacinol, and parietin were identified as major compounds in the tested extracts of the lichen. Parietin was isolated from the acetone extract on a separation column. The methanol extract had higher values of TPC (21.67 mg GAE/g) than the acetone extract. Isolated parietin showed the best antioxidant activity measures, according to the DPPH-scavenging activity assay (IC50 = 51.616 μg/mL) and reducing-power assay. Although the extracts showed the best antibacterial activity (especially against Proteus mirabilis ATCC 12453), parietin demonstrated superior antibiofilm activity (especially against Staphylococcus aureus ATCC 25923). This is the first report on the phytochemical composition of the lichen Placidium deosaiense and the first description of the chemical composition of some of the 45 species of the genus Placidium. This research will pave the way for further exploration of new activities of this lichen and its metabolites, which are important for medicine and pharmacy.

The Decontamination Effect of an Oscillating Chitosan Brush Compared With an Ultrasonic PEEK-Tip: An InVitro Study Using a Dynamic Biofilm Model.

This study aimed to assess the effect of an oscillating chitosan brush (OCB) compared with an ultrasonic device with PEEK tip (US-PEEK) for mechanical implant surface decontamination using an invitro model combining 3D models and a validated dynamic multispecies biofilm. A multispecies biofilm using six bacterial strains (Streptococcus oralis, Veillonella parvula, Actinomyces naeslundii, Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans) was seeded on dental implants with machined and sandblasted, large-grit and acid-etched (SLA) surfaces. These were installed in 3D models depicting peri-implant defect. Mechanical decontamination was performed for 120 s using either an OCB or a US-PEEK. A negative control group received no treatment. Scanning electron microscopy (SEM) was used to evaluate the bacterial composition and quantitative PCR (qPCR) analyzed the number of each bacterial species [colony-forming units per milliliter (CFU/mL)]. Well-structured biofilms with a dense microbial distribution were observed on the negative control implants after 72 h. qPCR following mechanical decontamination showed a scarce bacterial reduction in the OCB group. The US-PEEK group exhibited a significant decrease in bacterial species compared to both OCB and control groups (p < 0.05). A biofilm removal effect was also observed in the OCB group for the machined implant surfaces. In vitro assessment using an anatomical 3D model showed that mechanical decontamination effectively reduced biofilm. The US-PEEK group demonstrated biofilm reduction on the SLA surface, while the OCB group showed a reduction on the machined implant surface. Additionally, the US-PEEK group demonstrated greater efficacy in reducing bacterial numbers.

Quantitative evaluation of anti-biofilm cavitation activity seeded from microbubbles or protein cavitation nuclei by passive acoustic mapping

Bacterial biofilms represent a major challenge for effective antibiotic therapy as they confer physical and functional changes that protect bacteria from their surrounding environment. In this work, focused ultrasound in combination with cavitation nuclei was used to disrupt biofilms of Staphylococcus aureus and Pseudomonas aeruginosa, both of which are on the World Health Organization's priority list for new antimicrobial research. &#xD;Approach: Single species biofilms were exposed to ultrasound (0.5 MHz centre frequency, 0.5-1.5 MPa peak rarefactional pressure, 200 cycle pulses, 5 Hz repetition frequency, 30 s duration), in the presence of two different types of cavitation nuclei. Quantitative passive acoustic mapping (PAM) was used to monitor cavitation emissions during treatment using a calibrated linear array. &#xD;Main Results: It was observed that the cumulative energy of acoustic emissions during treatment was positively correlated with biofilm disruption, with differences between bacterial species attributed to differences in biofilm morphology. PCaN provided increased biofilm reduction compared to microbubbles due in large part to their persistence over the duration of ultrasound exposure. There was also good correlation between the spatial distribution of cavitation as characterized by PAM and the extent of biofilm disruption observed with microscopy. &#xD;Significance: Collectively, the results from this work indicate the potential broad applicability of cavitation for eliminating biofilms of priority pathogens and the opportunity presented by Passive Acoustic Mapping for real-time monitoring of antimicrobial processes.

Sublethal Damage Caused by Cold Plasma on Bacillus cereus Cells: Impact on Cell Viability and Biofilm-Forming Capacity.

The Bacillus cereus group represents a serious risk in powdered and amylaceous foodstuffs. Cold plasma (the fourth state of matter) is emerging as an alternative effective nonthermal technology for pasteurizing a wide range of matrices in solid, liquid, and powder form. The present study aims to evaluate the mechanisms involved in Bacillus cereus inactivation via cold plasma, focusing on (i) the technology's ability to generate damage in cells (at the morphological and molecular levels) and (ii) studying the effectiveness of cold plasma in biofilm mitigation through the direct effect and inhibition of the biofilm-forming capacity of sublethally damaged cells post-treatment. Dielectric barrier discharge cold plasma (DBD-CP) technology was used to inactivate B. cereus, B. thuringiensis, and B. mycoides under plasma power settings of 100, 200, and 300 W and treatment times ranging from 1 to 10 min. Inactivation levels were achieved in 2-7 log10 cycles under the studied conditions. Percentages of sublethally damaged cells were observed in a range of 45-98%, specifically at treatment times below 7 min. The sublethally damaged cells showed poration, erosion, and loss of integrity at the superficial level. At the molecular level, proteins and DNA leakage were also observed for B. cereus but were minimal for B. mycoides. Biofilms formed by B. cereus were progressively disintegrated under the DBD-CP treatment. The greater the CP treatment intensity, the greater the tearing of the bacteria's biofilm network. Additionally, cells sublethally damaged by DBD-CP were evaluated in terms of their biofilm-forming capacity. Significant losses in the damaged cells' biofilm network density and aggregation capacity were observed when B. cereus was recovered after inactivation at 300 W for 7.5 min, compared with the untreated cells. These results provide new insights into the future of tailored DBD-CP design conditions for both the inactivation and biofilm reduction capacity of B. cereus sensu lato species, demonstrating the effectiveness of cold plasma and the risks associated with sublethal damage generation.

Optimizing Desmostachya bipinnata-derived platinum nanoparticles for enhanced antibacterial and biofilm reduction

This study presents the green synthesis and comprehensive characterization of platinum nanoparticles (PtNPs) using Desmostachya bipinnata (Db) extract, incorporated into two innovative mouthwash formulations (MW1 and MW2). UV–Vis spectroscopy confirmed the successful synthesis of PtNPs, with distinct absorption peaks between 250 and 600 nm. Fourier-transform infrared (FTIR) spectroscopy identified hydroxyl and carbonyl functional groups, critical for the bioreduction and stabilization of PtNPs. High-resolution transmission electron microscopy (HR-TEM) revealed uniformly dispersed, spherical nanoparticles with a size range of 10–20 nm, while dynamic light scattering (DLS) confirmed a hydrodynamic diameter of 10–30 nm and a low polydispersity index (PDI) of 0.238, indicating excellent stability. Both formulations exhibited robust antimicrobial, antibiofilm, and anti-plaque properties, with MW2 showing superior efficacy, particularly against Staphylococcus aureus and Escherichia coli, as well as a notable 70 % reduction in biofilm formation and a 60 % plaque reduction within 2 h of treatment. The study underscores the potential of Desmostachya bipinnata-derived PtNPs as a promising alternative to conventional mouthwash, offering enhanced antimicrobial efficacy, biofilm disruption, and plaque prevention, alongside excellent stability and biocompatibility for oral healthcare applications.

The Hydrophobic Stabilization of Pseudomonas aeruginosa Bacteriophage F8 and the Influence of Modified Bacteriophage Preparation on Biofilm Degradation

The bacteriophage F8 belongs to the Myoviridae group of phages and is a pathogen of Pseudomonas aeruginosa. Since Pseudomonas aeruginosa is a multidrug-resistant opportunistic bacterium and can cause serious challenges for health services, studying the potential use of phages against them is a promising approach. Pseudomonas aeruginosa can be found on medical devices because bacteria can attach to surfaces and develop biofilms, which are difficult to eradicate because of their high resistance to environmental conditions and antimicrobial therapeutics. Phage therapy is becoming promising as an alternative for the treatment of antibiotic-resistant infections, but there is still a lack of standardized protocols approved by health organizations for possible use in the clinic. In our research, we focused on the potential use of 1-octanol, which was previously used by our team to develop a method for phage purification from bacterial lysate. 1-octanol is a fatty alcohol that is mostly used in the cosmetics industry, and its advantage is that it is approved by the FDA as a food additive. In this paper, we studied the protective properties of the addition of 1-octanol for storing phage liquid preparations. We demonstrated the stabilization effect of 1-octanol addition on F8 bacteriophage preparation during storage under various conditions. Interestingly, more effective biofilm reduction was observed after treatment with the purified bacteriophage and with 1-octanol addition compared to crude lysate.

A new type of Pseudomonas aeruginosa phage with potential as a natural food additive for eradicating biofilms and combating multidrug-resistant strains

Pseudomonas aeruginosa is a biofilm-forming bacterial pathogen with multidrug resistance that leads to serious foodborne illnesses. Bacteriophages as alternatives for controlling P. aeruginosa and its biofilms are of great interest, particularly in the food industry. This study reported, a new type of P. aeruginosa specific phage named vB PaeM HUST 1 and evaluated antibacterial and biofilm reduction to use alternative as an additive in food industry. vB PaeM HUST 1 showed excellent specificity and selectivity in host cell lysis and stability over a wide range of pH (3-10) and temperature (4–45 °C). Transmission electron microscopy revealed that the phage had a head with a long tail (head: 146.65 nm × 133.32 nm, tail: 199.98 nm); hence, it belongs to the subfamily Straboviridae. The 276114 bp vB PaeM HUST 1 genome encodes 360 ORFs, 7 tRNAs, and an average G + C content of 36.92%. In addition, they exhibited low lipopolysaccharide (LPS) levels at a concentration of 0.133 U/mL, with a phage titer of 1.14 × 109 PFU/mL and no cytotoxicity. vB PaeM HUST 1 efficiently degraded over 95% of the biofilm within 8 h and was highly effective against P. aeruginosa in both milk and beef, with significant reductions of 68.34% and 92% in bacterial concentration after 12 and 24 h of treatment, respectively, with complete eradication achieved after 36 h. Therefore, vB PaeM HUST 1 phage has unique characteristics and can be used as a novel additive to control P. aeruginosa in food matrices.

Alternating magnetic fields (AMF) and linezolid reduce Staphylococcus aureus biofilm in a large animal implant model

We aimed to evaluate the effectiveness of alternating magnetic fields (AMF) combined with antibiotics in reducing Staphylococcus aureus biofilm on metal implants in a large animal model, compared to antibiotics alone. Metal plates were inoculated with a clinical MRSA strain and then implanted into thirty-three ewes divided into three groups: positive control, linezolid only, and a combination of linezolid and AMF. Animals had either titanium or cobalt-chrome plates and were sacrificed at 5or 21 dayspost-implantation. Blood and tissue samples were collected at various time points post-AMF treatment. In vivo efficacy studies demonstrated significant biofilm reduction on titanium and cobalt-chrome implants with AMF-linezolid combination treatment compared to controls. Significant bacterial reductions were also observed in surrounding tissues and bones. Cytokine analysis showed improved inflammatory responses with combination therapy, and histopathology confirmed reduced inflammation, necrosis, and bacterial presence, especially at 5 days post-implantation. This study demonstrates that combining AMF with antibiotics significantly reduces biofilm-associated infections on metal implants in a large animal model. Numerical simulations confirmed targeted heating, and in vivo results showed substantial bacterial load reduction and reduced inflammatory response. These findings support the potential of AMF as a non-invasive treatment for prosthetic joint infections.

Combined antimicrobial activity of short peptide and phage-derived endolysin against antibiotic-resistant Salmonella Typhimurium

This study was designed to evaluate the combination effects of antimicrobial peptides (FK13 and FK16) and phage-encoded endolysin (LysPB32) on the inhibition of growth of polymyxin B-resistant Salmonella Typhimurium ATCC 19585 (STPMB). The inhibitory effects of FK13, FK16, and LysPB32 against STPMB were evaluated by using antimicrobial susceptibility, membrane permeability, biofilm reduction, cross-resistance, and mutant frequency assay. The minimum inhibitory concentrations (MICs) of FK13 and FK16 treated with LysPB32 (FK13+LysPB32 and FK16+LysPB32) against STPMB were decreased from more than 512 to 128 μg/ml and from 64 to 32 μg/ml, respectively. Compared to the control, the number of STPMB in the growing culture was reduced by 4.2 and 5.2 log CFU/ml, respectively, for FK13+LysPB32 and FK16+LysPB32 after 12-h incubation at 37 °C. All treatments (FK13, FK16, FK13+LysPB32, FK16+LysPB32) significantly increased the permeability of the outer membrane of STPMB. Biofilms were significantly decreased from OD600 of 0.6 to 0.16 for FK13+LysPB32 and from 0.6 to 0.13 for FK16+LysPB32. The ratios of MICs of erythromycin, ceftriaxone, polymyxin B, and ciprofloxacin to MIC of the control against STPMB were decreased to 0.50 for FK13+LysPB32 and FK16+LysPB32. The bactericidal activities of amikacin and gentamicin were enhanced for FK13+LysPB32 and FK16+LysPB32 (2-fold < MBC/MIC ratio). The mutant frequencies of STPMB to antibiotics were decreased when treated with FK13+LysPB32 and FK16+LysPB32. The results suggest that the combination of antimicrobial peptides and endolysins can be a promising strategy to control polymyxin B-resistant S. Typhimurium.

Combination of bacteriophages and vancomycin in a co-delivery hydrogel for localized treatment of fracture-related infections

Fracture-related infections (FRIs), particularly those caused by methicillin-resistant Staphylococcus aureus (MRSA), are challenging to treat. This study designed and evaluated a hydrogel loaded with a cocktail of bacteriophages and vancomycin (1.2 mg/mL). The co-delivery hydrogel showed 99.72% reduction in MRSA biofilm in vitro. The hydrogel released 54% of phages and 82% of vancomycin within 72 h and maintained activity for eight days, in vivo the co-delivery hydrogel with systemic antibiotic significantly reduced bacterial load by 0.99 log10 CFU compared to controls, with active phages detected in tissues at euthanasia (2 × 103 PFU/mL). No phage resistance was detected in the phage treatment groups, and serum neutralization resulted in only a 20% reduction in phage count. In this work, we show that a phage-antibiotic co-delivery system via CMC hydrogel is a promising adjunct to systemic antibiotic therapy for MRSA-induced FRI, highlighting its potential for localized, sustained delivery and improved treatment outcomes.

Are educational tools in oral health programs for pregnant women effective in improving the oral status of mothers and babies? A systematic review.

To analyze, through a systematic review, the effectiveness of educational tools (ETs) in Oral Health Programs (OHPs) for pregnant women in improving their oral status and thatof their babies. Searches were carried out in 5 electronic databases. Randomized (RCTs) and non-randomized clinical trials (CTs) were selected that included pregnant women exposed or not to ETs in OHPs, whose oral health status, in the pre- or postnatal period, or of their babies were verified. The types of ETs, outcomes, and results were extracted. Bias risk was assessed by ROBINS-I and RoB 2.0; and the certainty of the evidence (CE) by GRADE. A total of nine studies were included. Most used more than one ET (n = 8), with verbal guidance used in all (n = 9). Mothers showed a reduction in biofilm (n = 3), caries (n = 4) and periodontitis (n = 4). All studies with babies (n = 3) were successful in preventing caries. RCTs (n = 3) were classified as low risk of bias (n = 1), with concerns (n = 1) and high risk of bias (n = 1). CTs (n = 6) presented risk of bias as low (n = 2), serious (n = 1) and critical (n = 3). Studies about caries in babies showed high CE, and those who evaluated caries and periodontal parameters/biofilm in the mother had moderate and low CE, respectively. ETs as a strategy of OHPs for pregnant women can be effective in preventing caries in themselves and in their babies. However, despite the improvement of the mothers' oral hygiene and gingival condition, the CE was low considering this parameter.

Green synthesis of trimetallic CuO/Ag/ZnO nanocomposite using Ziziphus spina-christi plant extract: characterization, statistically experimental designs, and antimicrobial assessment

In this study, Ziziphus spina christi leaves was used to synthesize a trimetallic CuO/Ag/ZnO nanocomposite by a simple and green method. Many characterizations e.g. FTIR, UV–vis DRS, SEM–EDX, TEM, XRD, zeta-size analysis, and DLS, were used to confirm green-synthesized trimetallic CuO/Ag/ZnO nanocomposite. The green, synthesized trimetallic CuO/Ag/ZnO nanocomposite exhibited a spherical dot-like structure, with an average particle size of around 7.11 ± 0.67 nm and a zeta potential of 21.5 mV. An extremely homogeneous distribution of signals, including O (79.25%), Cu (13.78%), Zn (4.42%), and Ag (2.55%), is evident on the surface of green-synthetic nanocomposite, according to EDX data. To the best of our knowledge, this is the first study to effectively use an industrially produced green trimetallic CuO/Ag/ZnO nanocomposite as a potent antimicrobial agent by employing different statistically experimental designs. The highest yield of green synthetic trimetallic CuO/Ag/ZnO nanocomposite was (1.65 mg/mL), which was enhanced by 1.85 and 5.7 times; respectively, by using the Taguchi approach in comparison to the Plackett–Burman strategy and basal condition. A variety of assays techniques were utilized to evaluate the antimicrobial capabilities of the green-synthesized trimetallic CuO/Ag/ZnO nanocomposite at a 200 µg/mL concentration against multidrug-resistant human pathogens. After a 36-h period, the tested 200 µg/mL of the green-synthetic trimetallic CuO/Ag/ZnO nanocomposite effectively reduced the planktonic viable counts of the studied bacteria, Escherichia coli and Staphylococcus aureus, which showed the highest percentage of biofilm reduction (98.06 ± 0.93 and 97.47 ± 0.65%; respectively).

Nanotopography Influences Host-Pathogen Quorum Sensing and Facilitates Selection of Bioactive Metabolites in Mesenchymal Stromal Cells and Pseudomonas aeruginosa Co-Cultures.

Orthopedic implant-related bacterial infections and resultant antibiotic-resistant biofilms hinder implant-tissue integration and failure. Biofilm quorum sensing (QS) communication determines the pathogen colonization success. However, it remains unclear how implant modifications and host cells are influenced by, or influence, QS. High aspect ratio nanotopographies have shown to reduce biofilm formation of Pseudomonas aeruginosa, a sepsis causing pathogen with well-defined QS molecules. Producing such nanotopographies in relevant orthopedic materials (i.e., titanium) allows for probing QS using mass spectrometry-based metabolomics. However, nanotopographies can reduce host cell adhesion and regeneration. Therefore, we developed a polymer (poly(ethyl acrylate), PEA) coating that organizes extracellular matrix proteins, promoting bioactivity to host cells such as human mesenchymal stromal cells (hMSCs), maintaining biofilm reduction. This allowed us to investigate how hMSCs, after winning the race for the surface against pathogenic cells, interact with the biofilm. Our approach revealed that nanotopographies reduced major virulence pathways, such as LasR. The enhanced hMSCs support provided by the coated nanotopographies was shown to suppress virulence pathways and biofilm formation. Finally, we selected bioactive metabolites and demonstrated that these could be used as adjuncts to the nanostructured surfaces to reduce biofilm formation and enhance hMSC activity. These surfaces make excellent models to study hMSC-pathogen interactions and could be envisaged for use in novel orthopedic implants.

Metabolomic Approaches to Study the Potential Inhibitory Effects of Plantaricin Q7 against Listeria monocytogenes Biofilm.

Listeria monocytogenes is a serious pathogen and can exacerbate harmful effects through the formation of biofilm. Inhibition of or reduction in L. monocytogenes biofilm is a promising strategy to control L. monocytogenes in the food industry. In our previous study, it was found that plantaricin Q7 produced by Lactiplantibacillus plantarum Q7 could inhibit and reduce L. monocytogenes biofilm, but the specific mechanism remains unclear. In this study, the inhibitive and reduced activity of plantaricin Q7 on L. monocytogenes biofilm was investigated by metabolomics. The results showed that plantaricin Q7 inhibited the synthesis of L. monocytogenes biofilm mainly through purine metabolism and glycerol phospholipid metabolism, and the key differential metabolites included acetylcholine and hypoxanthine with a decrease in abundance from 5.80 to 4.85. In addition, plantaricin Q7 reduced the formed L. monocytogenes biofilm by purine metabolism and arginine biosynthesis, and the main differential metabolites were N-acetylglutamate and D-ribose-1-phosphate with a decrease in abundance from 6.21 to 4.73. It was the first report that purine metabolism and amino acid metabolism were the common metabolic pathway for plantaricin Q7 to inhibit and reduce L. monocytogenes biofilm, which could be potential targets to control L. monocytogenes biofilm. A putative metabolic pathway for L. monocytogenes biofilm inhibition and reduction by plantaricin Q7 was proposed. These findings provided a novel strategy to control L. monocytogenes biofilm in food processing.

Efficacy of a lytic bacteriophage vB_EcoM_SQ17 against Enterohemorrhagic Escherichia coli O157:H7 and Enterotoxigenic E. coli biofilms on cucumber

Enterohemorrhagic Escherichia coli O157:H7 (EHEC O157:H7) and Enterotoxigenic E. coli (ETEC) have been found to readily develop biofilms on cucumber (Cucumis sativus L.), presenting a significant risk to the safety of ready-to-eat vegetables. This study aimed to assess the effectiveness of the lytic bacteriophage vB_EcoM_SQ17 (SQ17) against EHEC O157:H7 and ETEC biofilms on cucumber. Here, we evaluated the efficacy of phage SQ17 on the formation and reduction of biofilms formed by EHEC O157:H7 and ETEC strains on various surfaces, including polystyrene, poly-d-lysine precoated films, and fresh-cut cucumber, at different temperatures. Phage SQ17 significantly inhibited ETEC biofilm formation, reducing the number of adhered cells by 0.15 log CFU/mL at 37 °C. Treatment with phage SQ17 also significantly decreased the number of adhered cells in established biofilms via SEM observation. Moreover, phage SQ17 effectively reduced the biomass of EHEC O157:H7 and ETEC biofilms by over 54.8 % at 37 °C after 24 h of incubation. Following phage treatment, the viability of adhered EHEC O157:H7 cells decreased by 1.37 log CFU/piece and 0.46 log CFU/piece in biofilms on cucumber at 4 °C and 25 °C, respectively. Similarly, the viability of ETEC cells decreased by 1.07 log CFU/piece and 0.61 log CFU/piece in biofilms on cucumber at 4 °C and 25 °C, respectively. These findings suggest that phage SQ17 shows promise as a potential strategy for eradicating pathogenic E. coli biofilms on cucumber.

Destruction of cariogenic biofilms by antimicrobial photodynamic therapy mediated by phycocyanin and toluidine blue along with sodium fluoride varnish or titanium tetrafluoride

ObjectiveEvaluation of the effect of phycocyanin (PC) and toluidine blue (TBO) along with sodium fluoride varnish (FV) or titanium tetrafluoride (TiF4) under the conditions of antimicrobial photodynamic therapy (PDT) on a dual-species cariogenic biofilm and on remineralization process. DesignAfter the development of Streptococcus mutans and Lactobacillus acidophilus dual-species biofilms on the human enamel disks, they were divided into 11 groups (n = 9): Control (0.9 % saline), PC, TBO, FV, and TiF4 alone, PC and TBO in combination with a 635 nm diode laser (PDT treatment), PC-PDT+ (PC + FV or TiF4 + 635 nm diode laser), and TBO-PDT+ (TBO + FV or TiF4 + 635 nm diode laser). After the treatment, crystal violet assay was performed to determine the reduction of cariogenic biofilms. Enamel remineralization changes were analyzed using energy dispersive X-ray spectroscopy (EDX) and field emission scanning electron microscopy (FESEM) for the calcium and phosphorus (Ca/P) ratio. ResultsOnly TBO-PDT+ showed superior antibiofilm activity when TiF4 was applied. Furthermore, the highest Ca/P ratio was found after treatment of enamel surfaces with TiF4-TBO-PDT+. The FESEM images showed that the enamel disks treated with TiF4 plus TBO-mediated PDT exhibited surface coating. However, TiF4 plus PC-mediated PDT cannot repair demineralized enamel. ConclusionsThese data suggest that TBO-PDT along with TiF4 can effectively reduce cariogenic biofilms and significantly remineralize enamel disks, opening new avenues in caries prevention.

Effect of Novel and Traditional Intracanal Medicaments on Biofilm Viability and Composition

IntroductionThe aim of this study was to test the hypothesis that a combination of D-amino acids (DAAs) and trans-cinnamaldehyde (TC) demonstrates superior antibiofilm activity to calcium hydroxide (CH) and untreated controls. MethodsIn this 3-part in vitro study, the concentration of DAAs (D-methionine, D-leucine, D-tyrosine, and D-tryptophan) that would significantly decrease Enterococcus faecalis and Actinomyces naeslundii biofilm biomass was first determined. Then, the effect of TC + selected DAAs on polymicrobial biofilms was characterized by quantifying the biomass and biofilm viability. Finally, the antibiofilm effects of TC + DAA was compared with CH and untreated controls by (i) determining bacterial viability and (ii) quantifying biofilm matrix composition using selective fluorescence-binding analysis. Statistical analysis was performed using one-way ANOVA and appropriate multiple comparisons test, with P < .05 considered as statistically significant. ResultsTC (0.06%) + D-tyrosine (1 mM) + D-tryptophan (25 mM) significantly reduced the biomass and biofilm viability compared to the control (P < .05). While no significant difference was observed between TC + DAA and CH in the cultivable bacterial counts (P > .05), confocal microscopy demonstrated a significantly greater percentage of dead bacteria in TC + DAA-treated biofilms compared to CH and the control (P < .05). TC + DAA significantly decreased the biovolume and all the examined components of the biofilm matrix quantity compared to the control, while CH significantly reduced only the exopolysaccharide quantity (P < .05). ConclusionThe combination of TC + D-tyrosine + D-tryptophan demonstrated superior antibiofilm activity (biofilm bacterial killing and reduction of matrix quantity) to CH and has potential to be developed as an intracanal medicament.