In response to the global rise in antibiotic resistance and the prevalence of bacterial biofilm-related infections, the antibacterial efficacy of methanolic, ethanolic, and aqueous extracts of 18 Lamiaceae plants from Serbia was evaluated. The total coumarins and triterpenes were detected spectrophotometrically, while a microdilution assay measured their effects on bacterial growth. Additionally, the impact of these extracts was assessed on Pseudomonas aeruginosa PAO1 adhesion and invasion in human fibroblasts and biofilm formation and degradation. The alcoholic extracts had the highest phytochemical content, with Teucrium montanum and Lavandula angustifolia being the richest in coumarins and triterpenes, respectively. Gram-positive bacteria, particularly Bacillus subtilis, were more susceptible to the extracts. Hyssopus officinalis ethanolic and Sideritis scardica methanolic extracts inhibited bacterial growth the most efficiently. Although the extracts did not inhibit bacterial adhesion, most ethanolic extracts significantly reduced bacterial invasion. Origanum vulgare and H. officinalis ethanolic extracts significantly inhibited biofilm formation, while Teucrium chamaedrys extract was the most active in biofilm degradation. This study significantly contributes to the literature by examining the antibacterial activity of Lamiaceae extracts, addressing major literature gaps, and underscoring their antibacterial potential, particularly Satureja montana and O. vulgare ethanolic extracts, linking their efficacy to coumarins and triterpenes.

In nature, bacteria are ubiquitous and can be categorized as beneficial or harmless to humans, but most bacteria have one thing in common which is their ability to produce biofilm. Biofilm is encased within an extracellular polymeric substance (EPS) which provides resistance against antimicrobial agents. Protease enzymes have the potential to degrade or promote the growth of bacterial biofilms. In this study, the effects of a recombinant intracellular serine protease from Bacillus sp. (SPB) on biofilms from Staphylococcus aureus, Acinetobacter baumannii, and Pseudomonas aeruginosa were analyzed. SPB was purified using HisTrap HP column and concentrated using Amicon 30 ultra-centrifugal filter. SPB was added with varying enzyme activity and assay incubation period after biofilms were formed in 96-well plates. SPB was observed to have contrasting effects on different bacterial biofilms, where biofilm degradations were observed for both 7-day-old A. baumannii (37.26%) and S. aureus (71.51%) biofilms. Meanwhile, SPB promoted growth of P. aeruginosa biofilm up to 176.32%. Compatibility between protein components in S. aureus biofilm with SPB as well as a simpler membrane structure morphology led to higher biofilm degradation for S. aureus compared to A. baumannii. However, SPB promoted growth of P. aeruginosa biofilm due likely to its degrading protein factors that are responsible for biofilm detachment and dispersion, thus resulting in more multi-layered biofilm formation. Commercial protease Savinase which was used as a comparison showed degradation for all three bacterial biofilms. The results obtained are unique and will expand our understanding on the effects that bacterial proteases have toward biofilms.

Bacterial biofilms have attracted significant attention due to their involvement in persistent infections, food and water contamination, and infrastructure corrosion. This review delves into the intricate interactions between bacterial biofilms and unicellular parasites, shedding light on their impact on biofilm formation, structure, and function. Unicellular parasites, including protozoa, influence bacterial biofilms through grazing activities, leading to adaptive changes in bacterial communities. Moreover, parasites like Leishmania and Giardia can shape biofilm composition in a grazing independent manner, potentially influencing disease outcomes. Biofilms, acting as reservoirs, enable the survival of protozoan parasites against environmental stressors and antimicrobial agents. Furthermore, these biofilms may influence parasite virulence and stress responses, posing challenges in disease treatment. Interactions between unicellular parasites and fungal-containing biofilms is also discussed, hinting at complex microbial relationships in various ecosystems. Understanding these interactions offers insights into disease mechanisms and antibiotic resistance dissemination, paving the way for innovative therapeutic strategies and ecosystem-level implications.

Dental biofilms represent a serious oral health problem playing a key role in the development of caries and other oral diseases. In the present work, we cloned and expressed in E. coli two glucanases, Prevotella melaninogenica mutanase (PmGH87) and Capnocytophaga ochracea dextranase (CoGH66), and characterized them biochemically and biophysically. Their three-dimensional structures were elucidated and discussed. Furthermore, we tested the capacity of the enzymes to hydrolyze mutan and dextran to prevent formation of Streptococcus mutans biofilms, as well as to degrade pre- formed biofilms in low and abundant sugar conditions. The percentage of residual biofilm was calculated for each treatment group in relation to the control, as well as the degree of synergism. Our results suggest that both PmGH87 and CoGH66 are capable of inhibiting biofilm formation grown under limited or abundant sucrose conditions. Degradation of pre-formed biofilms experiments reveal a time-dependent effect for the treatment with each enzyme alone. In addition, a synergistic and dose-dependent effects of the combined enzymatic treatment with the enzymes were observed. For instance, the highest biomass degradation was 95.5% after 30 min treatment for the biofilm grown in low sucrose concentration, and 93.8% after 2 h treatment for the biofilm grown in sugar abundant condition. Strong synergistic effects were observed, with calculated degree of synergism of 5.54 and 3.18, respectively and their structural basis was discussed. Jointly, these data can pave the ground for the development of biomedical applications of the enzymes for controlling growth and promoting degradation of established oral biofilms.

The emergence, spread and difficult removal of bacteria biofilm, represent an ever-increasing persistent infections and medical complications challenge worldwide. Herein, a self-propelled system Prussian blue micromotor (PB MMs) were constructed by gas-shearing technology for efficient degradation of biofilms by combining chemodynamic therapy (CDT) and photothermal therapy (PTT). With the interpenetrating network crosslinked by alginate, chitosan (CS) and metal ions as the substrate, PB was generated and embedded in the micromotor at the same time of crosslinking. The micromotors are more stable and could capture bacteria with the addition of CS. The micromotors show excellent performance, containing photothermal conversion, reactive oxygen species (ROS) generation and bubble produced by catalyzing Fenton reaction for motion, which served as therapeutic agent could chemically kill bacteria and physically destroy biofilm. This research work opens a new path of an innovative strategy to efficiently remove biofilm.

In the field of phage applications and clinical treatment, virulent phages have been in the spotlight whereas temperate phages received, relatively speaking, less attention. The fact that temperate phages often carry virulent or drug-resistant genes is a constant concern and drawback in temperate phage applications. However, temperate phages also play a role in bacterial regulation. This review elucidates the biological properties of temperate phages based on their life cycle and introduces the latest work on temperate phage applications, such as on host virulence reduction, biofilm degradation, genetic engineering and phage display. The versatile use of temperate phages coupled with their inherent properties, such as economy, ready accessibility, wide variety and host specificity, make temperate phages a solid candidate in tackling bacterial infections.

Cutibacterium acnes (C. acnes) is the main causative agent of acne vulgaris. The study aims to evaluate the antimicrobial activity of a natural product, Arctostaphylos uva-ursi leaf extract, against C. acnes. Preliminary chemical-physical characterization of the extract was carried out by means of FT-IR, TGA and XPS analyses. Skin permeation kinetics of the extract conveyed by a toning lotion was studied in vitro by Franz diffusion cell, monitoring the permeated arbutin (as the target component of the extract) and the total phenols by HPLC and UV-visible spectrophotometry, respectively. Antimicrobial activity and time-killing assays were performed to evaluate the effects of Arctostaphylos uva-ursi leaf extract against planktonic C. acnes. The influence of different Arctostaphylos uva-ursi leaf extract concentrations on the biofilm biomass inhibition and degradation was evaluated by the crystal violet (CV) method. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test was used to determine the viability of immortalized human keratinocytes (HaCaT) after exposure to Arctostaphylos uva-ursi leaf extract for 24 and 48 h. Levels of interleukin (IL)-1β, IL-6, IL-8 and tumour necrosis factor (TNF)-α were quantified after HaCaT cells cotreatment with Arctostaphylos uva-ursi leaf extract and heat-killed C. acnes. The minimum inhibitory concentration (MIC) which exerted a bacteriostatic action on 90% of planktonic C. acnes (MIC90) was 0.6 mg/mL. Furthermore, MIC and sub-MIC concentrations influenced the biofilm formation phases, recording a percentage of inhibition that exceeded 50 and 40% at 0.6 and 0.3 mg/mL. Arctostaphylos uva-ursi leaf extract disrupted biofilm biomass of 57 and 45% at the same concentrations mentioned above. Active Arctostaphylos uva-ursi leaf extract doses did not affect the viability of HaCaT cells. On the other hand, at 1.25 and 0.6 mg/mL, complete inhibition of the secretion of pro-inflammatory cytokines was recorded. Taken together, these results indicate that Arctostaphylos uva-ursi leaf extract could represent a natural product to counter the virulence of C. acnes, representing a new alternative therapeutic option for the treatment of acne vulgaris.

To investigate the degradation effect of bovine trypsin on multispecies biofilm of periodontitis-related bacteria and to provide an experimental reference for exploring new methods for controlling biofilms of periodontitis-related microorganisms, the multispecies biofilm of periodontitis-related microorganisms was established. Standard strains of Porphyromonas gingivalis, Fusobacterium nucleatum subsp. polymorpha, Actinomyces viscosus, and Aggregatibacter actinomycetemcomitans were co-cultured to form the biofilm. The experimental groups were treated with bovine trypsin, distilled water was applied as the blank control group, and phosphate saline buffer (pH = 7.4) as the negative control group. Morphological observation and quantitative analysis of extracellular polymeric substances (EPS), live bacteria, and dead bacteria were conducted using a laser confocal microscope. The morphological changes of EPS and bacteria were also observed using a scanning electron microscope. The results of morphological observations of modeling were as follows. EPS aggregated as agglomerates, and bacteria flora were wrapped by them, showing a three-dimensional network structure, and channel-like structures were inside the biofilm. Live bacteria were distributed on the surface of the EPS or embedded in them, dead bacteria aggregated between live flora and the bottom layer of biofilms. After being treated with bovine trypsin, the three-dimensional network structure and the channel-like structure disappeared, and the EPS and live and dead bacteria decreased. Quantitative analysis results are as follows. When biofilm was treated for 30 s, 1 min, and 3 min, the minimum effective concentrations of bovine trypsin to reduce EPS were 2 mg/ml (P < 0.05), 0.5 mg/ml (P < 0.05), and 0.25 mg/ml (P < 0.05), respectively. The minimum effective concentrations of bovine trypsin to reduce the live or dead bacteria were 2 mg/ml (P < 0.05), 0.5 mg/ml (P < 0.05), and 0.5 mg/ml (P < 0.05), respectively. There was no significant difference in the ratio of live/dead bacteria after the biofilm was treated for 30 s with bovine trypsin at the concentration of 0.25, 0.5, 1, and 2 mg/ml (P > 0.05), and the minimum effective concentration to reduce the ratio of live bacteria/dead bacteria was 0.25 mg/ml (P < 0.05) after treatment for 1 min and 3 min. Therefore, bovine trypsin can destroy biofilm structure, disperse biofilm and bacteria flora, and reduce the EPS and bacterial biomass, which are positively correlated with the application time and concentration.

The aim of the study was to assess the impact of four unifloral honeys on the food-borne pathogens Pseudomonas aeruginosa and Staphylococcus aureus, by analyzing the honeys\' antibacterial and biofilm degradation effects, as well as their antioxidant activity and element content. Linden and milkweed honeys represented light colored honeys, while goldenrod and chestnut honeys the darker ones. The botanical origin of the honeys and the relative frequency of their pollen types were established with melissopalynological analysis. The antioxidant capacities were calculated by two single electron transfer based methods (TRC - Total Reducing Capacity and TEAC - Trolox Equivalent Antioxidant Capacity) and a hydrogen atom transfer based assay (ORAC - Oxygen Radical Absorbance). The amount of four main macro- and two microelements was quantified. The antibacterial activity was determined by minimum inhibitory concentration (MIC) and membrane degradation assays. Furthermore, the biofilm degradation power of the samples was studied as well. The light colored linden honey with the lowest TRC and TEAC, but with the highest ORAC antioxidant activity and high element content showed the best antibacterial and biofilm degradation effects. Meanwhile, the dark colored chestnut honey with significantly higher single electron transfer based antioxidant capacities, with high element content, but lower ORAC showed significantly higher MIC and lower membrane degradation activity than linden honey. In case of biofilm degradation, both honey types gave similarly high inhibitory effect. Goldenrod honey was similarly effective regarding its MIC properties like chestnut honey, but had significantly lower antioxidant potential and ability to disrupt bacterial membranes and biofilms. Milkweed honey was the honey type with the lowest bioactivity and element content. The honeys, unequivocally characterized by their antioxidant characters and element content, displayed different antibacterial and biofilm degradation effects. In addition, some honey traits were found to be good predictors of the antimicrobial potential of honeys: ORAC assay showed correlation with the MIC values of both bacteria, and strict correlation was found between the mineral content and the antibiofilm activity of the studied honeys. Our studies indicate that unifloral honeys, such as linden and chestnut honeys, are plant-derived products with great potential as antimicrobial agents in food preservation, exhibiting remarkable antibacterial activity against food-borne pathogens.

OBJECTIVE: The degradation activity of two bacteriophages UPMK_1 and UPMK_2 against methicillin-resistant Staphylococcus aureus phages were examined using gel zymography.
METHODS: The analysis was done using BLASTP to detect peptides catalytic domains. Many peptides that are related to several phage proteins were revealed.
RESULTS: UPMK_1 and UPMK_2 custom sequence database were used for peptide identification. The biofilm-degrading proteins in the bacteriophage UPMK_2 revealed the same lytic activity towards polysaccharide intercellular adhesin-dependent and independent of Methicillin-resistant Staphylococcus aureus (MRSA) biofilm producers in comparison to UPMK_1, which had lytic activity restricted solely to its host.
CONCLUSIONS: Both bacteriophage enzymes were involved in MRSA biofilm degradation during phage infection and they have promising enzybiotics properties against MRSA biofilm formation.