Efflux pump inhibitors are a potential therapeutic strategy for managing antimicrobial resistance and biofilm formation. This article evaluated the effect of carbonyl cyanide m-chlorophenyl hydrazone (CCCP) on the biofilm growth dynamics and the production of virulence factors by Burkholderia pseudomallei. The effects of CCCP on planktonic, growing, and mature biofilm, interaction with antibacterial drugs, and protease and siderophore production were assessed. CCCP MICs ranged between 128 and 256 µM. The CCCP (128 µM) had a synergic effect with all the antibiotics tested against biofilms. Additionally, CCCP reduced (p < .05) the biomass of biofilm growth and mature biofilms at 128 and 512 µM, respectively. CCCP also decreased (p < .05) protease production by growing (128 µM) and induced (p < .05) siderophore release by planktonic cells (128 µM) growing biofilms (12.8 and 128 µM) and mature biofilms (512 µM). CCCP demonstrates potential as a therapeutic adjuvant for disassembling B. pseudomallei biofilms and enhancing drug penetration.
Using existing adrentimicrobials with essential oil components to prevent antimicrobial resistance is an alternative strategy. This study aimed to evaluate the resistance status, synergistic combinations, and in vitro biofilm formation activities of clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA), Stenotrophomonas maltophilia and Candida albicans against antimicrobial agents and cinnamaldehyde, carvacrol, eugenol, limonene and eucalyptol. Antimicrobial activities were evaluated by microdilution, cytotoxicity by XTT, synergy by checkerboard and time-kill, and biofilm inhibition by microplate methods. Cinnamaldehyde and carvacrol showed strong antimicrobial activity. Synergistic effects were observed when using all essential oils with antimicrobials. Only two C. albicans isolates showed antagonism with cinnamaldehyde and fluconazole. The constituents showed cytotoxic effects in the L929 cell line (except limonene). A time-kill analysis revealed a bacteriostatic effect on S. maltophilia and MRSA isolates and a fungicidal effect on C. albicans isolates. These results are important for further research to improve antimicrobial efficacy or to develop new agents.
Polyethylene terephthalate (PET) and polylactic acid (PLA) are among the polymers used in the food industry. In this study, crude extracts of Dunaliella salina were used to treat the surface of 3D printed materials studied, aiming to provide them with an anti-adhesive property against Pseudomonas aeruginosa. The hydrophobicity of treated and untreated surfaces was characterized using the contact angle method. Furthermore, the adhesive behavior of P. aeruginosa toward the substrata surfaces was also studied theoretically and experimentally. The results showed that the untreated PLA was hydrophobic, while the untreated PET was hydrophilic. It was also found that the treated materials became hydrophilic and electron-donating. The total energy of adhesion revealed that P. aeruginosa adhesion was theoretically favorable on untreated materials, while it was unfavorable on treated ones. Moreover, the experimental data proved that the adhesion to untreated substrata was obtained, while there was complete inhibition of adhesion to treated surfaces.
Understanding factors influencing Listeria monocytogenes biofilms aid in developing more effective elimination/prevention strategies. This study examined the effect of temperature (4 °C, 21 °C, 30 °C), materials (stainless steel 316 L with 2B and 2 R finishes, glass, and polypropylene), and slope (0°/horizontal or 90°/vertical) on mono- and dual-species biofilms using two L. monocytogenes strains and one Pseudomonas aeruginosa strain. All biofilms were grown in 10% TSB for 24 h and analyzed using culture-based methods. Additionally, the architecture of monospecies biofilms was studied using fluorescence microscopy. Overall, P. aeruginosa showed higher biofilm formation potential (6.2 log CFU/cm2) than L. monocytogenes (4.0 log CFU/cm2). Temperature greatly influenced P. aeruginosa and varied for L. monocytogenes. The slope predominantly influenced L. monocytogenes monospecies biofilms, with cell counts increasing by up to 2 log CFU/cm2. Surface material had little impact on biofilm formation. The study highlights the varying effects of different parameters on multispecies biofilms and the importance of surface geometry.
The bacterial community from a cooling water system was investigated through culture-dependent and independent strategies, and the responses of planktonic and sessile bacteria (grown in glass slides and stainless-steel coupons) to antimicrobials of industrial and clinical use were assessed. The morphotypes with higher biofilm-forming potential were Pseudoxanthomonas sp., Rheinheimera sp., Aeromonas sp. and Staphylococcus sp., and the first also exhibited lower susceptibility to all antibiotics and biocides tested. 16S rRNA high throughput sequencing indicated that Pseudomonadota (77.1% on average, sd 11.1%), Bacteroidota (8.4, sd 5.7%), and Planctomycetota (3.0, sd 1.3%) were the most abundant phyla. KEGG orthologs associated with antibiotics and biocide resistance were abundant in all samples. Although the minimum inhibitory and bactericidal concentrations were generally higher for biofilms, morphotypes in planktonic form also showed high levels of resistance, which could be associated with biofilm cells passing into the planktonic phase. Overall, monochloramine was the most effective biocide.
Microbial fouling involves the physicochemical interactions between microorganisms and solid surfaces. An electromagnetic field (EMF) may change the diffusion rates of microbial cells and the electrical double layer around the cells and contacting surfaces. In the current study, polycardanol exhibiting antibiofouling activity was modified with ferromagnetic iron oxide (IO) to investigate the EMF effects on bacterial adhesion. When there was a flow of electrolyte that contained bacterial cells, flow-induced EMF was generated according to Faraday's principle. It was observed that the IO-ionic solution (IS)-modified surfaces, with an induced current of 44, 53, 66 nA, showed decreases in the adhesion of bacteria cells more than the unmodified (polycardanol) and IO-nanoparticles-modified ones. In addition to the EMF effects, the nano-scale uniform roughness of the modified surfaces appeared to play an important role in the reduction of cell adhesion. The results demonstrated that the IOIS-modified surface (3.2 × 10-6 mM IO) had the highest antibiofouling activity.
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.
Artificial reefs represent useful tools to revitalize coastal and ocean ecosystems. Their formulation determines the biofilm formation which is the prerequisite for the colonization process by marine micro- and macroorganisms. In comparison with concrete, biobased polymers offer improved characteristics, including architecture, formulation, rugosity and recycling. This article aims to explore a new scale of artificial reef made of biocomposites reinforced with a high flax fibre (Linum utilatissimum) content (30%). Cellular adhesion and resulting biofilm formation were assessed using two marine microorganisms: Pseudoalteromonas sp. 3J6 and Cylindrotheca closterium. The influence of flax fibre leachates and plastic monomers on the growth of those marine microorganisms were also evaluated. Results indicated that the introduction of flax fibres inside the polymer matrix modified its physicochemical properties thus modulating adhesion and biofilm formation depending on the microorganism. This study gives insights for further developments of novel functionalized artificial reefs made of biocomposites.
Biofouling on marine surfaces causes immense material and financial harm for maritime vessels and related marine industries. Previous reports have shown the effectiveness of amphiphilic coating systems based on poly(dimethylsiloxane) (PDMS) against such marine foulers. Recent studies on biofouling mechanisms have also demonstrated acidic microenvironments in biofilms and stronger adhesion at low-pH conditions. This report presents the design and utilization of amphiphilic polymer coatings with buffer functionalities as an active disruptor against four different marine foulers. Specifically, this study explores both neutral and zwitterionic buffer systems for marine coatings, offering insights into coating design. Overall, these buffer systems were found to improve foulant removal, and unexpectedly were the most effective against the diatom Navicula incerta.