Search Results (67)

Polymicrobial biofilms involving fungal and bacterial species are increasingly recognized as contributors to persistent infections, particularly in the oral cavity. Candida albicans and Aggregatibacter actinomycetemcomitans are two commensals that can turn into opportunistic pathogens and are able to form robust biofilms. Objectives: This study aimed to assess the interaction dynamics between these two microorganisms and to evaluate their susceptibility to fluconazole and azithromycin in single- and mixed-species forms. Methods: Biofilm biomass was quantified using crystal violet assays, while biofilm cell viability was assessed through CFU enumeration (biofilm viability assay). To assess the resistance properties of single versus mixed-species coincubations, we applied the antimicrobial susceptibility test (AST) to each drug, and analysed spatial organization with confocal laser scanning microscopy, using PNA-FISH. Results: The results indicated that both species can coexist without significant mutual inhibition. However, a non-reciprocal synergism was also observed, whereby mixed-species biofilm conditions promoted the growth of A. actinomycetemcomitans, while C. albicans growth remained stable. As expected, antimicrobial tolerance was elevated in mixed cultures, likely due to enhanced extracellular matrix production and potential quorum-sensing interactions, contributing to increased resistance against azithromycin and fluconazole. Conclusions: This study provides novel insights into previously rarely explored interactions between C. albicans and A. actinomycetemcomitans. These findings underscore the importance of investigating interspecies interactions within polymicrobial biofilms, as understanding their mechanisms, such as quorum-sensing molecules and metabolic cooperation, can contribute to improved diagnostics and more effective targeted therapeutic strategies against polymicrobial infections. Full article

The prevention of biofilm formation and algal biodeterioration on building materials, particularly on cultural heritage sites, is a growing concern. Due to regulatory restrictions on conventional algicidal biocides in Europe, natural alternatives such as essential oils are gaining interest for their potential use in heritage conservation. This study evaluates the anti-algal activity of Salvia officinalis and Equisetum arvense (essential oils, hydrolates, and extracts) against a mixed culture of five green algae species (Bracteacoccus minor, Stichococcus bacillaris, Klebsormidium nitens, Chloroidium saccharophilum, and Diplosphaera chodatii). The plant materials were processed using hydrodistillation and solvent extraction, followed by chemical characterization through gas chromatography–mass spectrometry (GC-MS). Biological efficacy was assessed by measuring algal growth inhibition, changes in biomass colour, chlorophyll a concentration, and fluorescence. S. officinalis yielded higher extract quantities (extraction yield: 23%) than E. arvense and contained bioactive compounds such as thujone, camphor, and cineole, which correlated with its strong anti-algal effects. The essential oil of S. officinalis demonstrated the highest efficacy, significantly inhibiting biofilm formation (zones of inhibition: 15–94 mm) and photosynthetic activity at 0.5% concentration (reduction in chlorophyll a concentration 90–100%), without causing visible discolouration of treated surfaces (∆E < 2). These findings highlight the potential of S. officinalis essential oil as a natural, effective, and material-safe algicidal biocide for the sustainable protection of cultural heritage sites. Full article

Fungal prosthetic joint infection (fPJI) is one of the orthopaedic pathologies where there is no clear evidence, guidelines or algorithm to guide the surgeon in its management. This is in addition to the difficulty with which these infections are diagnosed, isolated and treated. Fungi form notorious biofilms that are difficult to eradicate once formed and that display resistance to antimicrobial agents. These biofilms have been shown to act synergistically with biofilms of bacteria, further adding to medical treatment resistance. We have reviewed the literature for reports that describe the results of different methods in surgically treating fPJI. We found that surgical management with two stages remains the gold standard for treatment of fPJI, as is the case for bacterial PJI (bPJI). We have investigated medical treatment, debridement with implant retention (DAIR) and staged revisions and whether a reasonable recommendation can be made based on the best knowledge and practice available. From the data on bPJI, there exists a role for conservative management of acute PJI with debridement, antibiotics and implant retention (DAIR). While fPJI and bPJI both represent infections, the differences in our ability to detect these infections clinically, culture the pathogens and treat them with proper antimicrobial agents, along with the difference in the reported results of the surgical treatment, make us believe that these two types of infections should not be treated in the same manner. With all this in mind, we reviewed several reports in the literature on fPJI to determine the efficacy of current treatment modalities, including DAIR, which followed current guidelines for PJI. Data show an overall treatment success rate of 64.4% [range 17.4–100%]. Subgroup analysis revealed a success rate of 11.6% [range 0–28.7%] in patients treated with DAIR. There is no doubt that DAIR should not be encouraged as it consistently has a bad record. Although there are not enough studies or numbers of patients to show an evidence-based preference over one- or two-staged revisions, the two-stage revision of fPJI consistently shows better results and should be considered as the gold standard of management in cases of revision fPJI. This should also be coupled with proper expertise, follow-ups and recommended lengths of medical treatment, which should not be less than six months. From the review of these data, we have developed reasonable recommendations for the management of fPJI. These recommendations center on staged surgical debridement along with medical management. Medical treatment should be for at least 6 months under the guidance of an infectious disease team and based on intraoperative cultures. In the case of local antimicrobial treatment reported in the literature, many patients with fPJI were found to have a polymicrobial infection. As a result, it is our recommendation that antifungals as well as antibacterials should be incorporated into the cement spacer mix of these cases. Fungal PJI remains an exceedingly difficult pathology to treat and should be managed by experienced surgeons in a well-equipped institution. Full article

Furfural is a relevant industrial product, but its presence in water and soil generates contamination and health risks. Moving bed biofilm reactors (MBBRs) are an increasingly used alternative to eliminate contaminants with the advantage of occupying small spaces, despite their high dependence on support and the microorganisms involved in the process. This work proposes furfural elimination through a laboratory-scale MBBR using Bacillus licheniformis GTQ1, Microbacterium sp. GISTAQ2, and Brevundimonas sp. GISTAQ1 isolated from an industrial effluent and agroforestry waste (rice husks, pine sawdust, and quebracho chips) as supports. The biofilm development was tested with both axenic and mixed cultures, confirming high coverage by Scanning Electron Microscope (SEM) images, especially in triple-mixed cultures. Biodegradation tests were carried out in the MBBR with 15 g rice husks or quebracho chips as supports and a 4000 mg L⁻¹ initial furfural concentration for 72 h. The mixed culture achieved almost a 100% furfural removal in three days with a rate of 3.97% per hour with rice husks and 2.61% per hour with quebracho chips. This laboratory-scale MBBR development is a promising first step ready for a scale-up for its implementation in industries to significantly reduce the environmental impact of the discharge of this type of effluent. Full article

This study introduces innovative varnishes incorporating natural bioactive compounds to inhibit the formation of oral dual biofilms, a critical contributor to dental caries and other oral diseases. The purpose of this study was to evaluate the effectiveness of bioactive varnishes containing tt-farnesol, quercetin, and theobromine in inhibiting the formation of mixed Streptococcus mutans and Candida albicans biofilms. Mixed biofilms of Streptococcus mutans UA159 and Candida albicans SC5314 were grown in 96-well plates containing a specialized culture medium. Approximately 0.2 mL of experimental varnishes with A—1.5% or B—4.5% concentrations of tt-farnesol, quercetin, and theobromine were separately added to the wells using a disposable applicator, with a vehicle varnish (lacking bioactives) serving as the control. Biofilms were incubated at 37 °C with 5% CO₂ for 24 h. Microbial viability was determined in terms of colony-forming units per milliliter (CFU/mL), and biofilm morphology was evaluated qualitatively via scanning electron microscopy (SEM). Statistical analyses were performed using ANOVA/Tukey tests at a 5% significance level. Varnishes A and B achieved significant reductions in microbial populations within the biofilms (p < 0.05) compared to the vehicle control (C). SEM imaging revealed marked structural disruptions in the biofilms, validating the quantitative results. Higher bioactive concentrations demonstrated enhanced inhibitory effects. Bioactive varnishes enriched with theobromine, quercetin, and tt-farnesol represent a novel and effective strategy for inhibiting oral dual biofilm development, offering a promising advancement in preventive dentistry. Full article

Campylobacter spp. are prevalent foodborne bacterial enteric pathogens. Their inclusion in biofilms on abiotic surfaces is considered a strategy that facilitates their extraintestinal survival. Organic acid (OA) treatments could be used in a green approach to decontaminate various surfaces. This work aimed to evaluate the inhibitory and eradicative effects of L(+)-lactic acid (LA), a naturally occurring OA, on a dual-species biofilm formed on two food processing model surfaces (polystyrene and stainless steel) by three selected foodborne Campylobacter spp. isolates (two C. jejuni and one C. coli). The influence of aerobiosis conditions (microaerophilic, aerobic and CO₂ enriched) on the resistance of the established biofilms to the acid was also tested. In parallel, the predominant metabolites contained in the planktonic media of biofilm monocultures and mixed-culture biofilm were comparatively analyzed by an untargeted metabolomics approach. Results revealed that LA inhibited mixed-culture biofilm formation by more than 2 logs (>99%) on both surfaces when this was applied at its highest tested concentration (4096 μg/mL; 0.34% v/v). However, all the preformed mixed-culture biofilms (ca. 10⁶⁻⁷ CFU/cm²) could not be eradicated even when the acid was used at concentrations exceeding 5% v/v, denoting their extremely high recalcitrance which was still influenced by the abiotic substratum, and the biofilm-forming aerobiosis conditions. The metabolic analysis revealed a strain-specific metabolite production which might also be related to the strain-specific biofilm-forming and resistance behaviors and resulted in the distinct clustering of the different samples. Overall, the current findings provide important information on the effectiveness of LA against biofilm campylobacteria and may assist in mitigating their risk in the food chain. Full article

This study explores the efficient decolorization and complete mineralization of the diazo dye Evans blue, using an integrated aerobic bioreactor system coupled with a double-chamber microbial fuel cell (DCMFC) including a bio-cathode and acetate as a cosubstrate. The research addresses the environmental challenges posed by dye-laden industrial effluents, focusing on achieving high decolorization efficiency and understanding the microbial communities involved. The study utilized mixed strains of actinomycetes, isolated from garden compost, to treat initial dye concentrations of 100 mg/L and 200 mg/L. Decolorization efficiency and microbial community composition were evaluated using 16S rRNA sequencing, and electrochemical impedance spectroscopy (EIS) was used to assess anode and DCMFC resistance. The results demonstrated decolorization efficiencies ranging from 90 ± 2% to 98 ± 1.9% for 100 mg/L and from 79 ± 2% to 87% ± 1% for 200 mg/L. An anode resistance of 12.48 Ω indicated a well-developed biofilm and enhanced electron transfer. The microbial community analysis revealed a significant presence of Pseudomonadota (45.5% in dye-acclimated cultures and 32% in inoculum cultures), with key genera including Actinomarinicola (13.75%), Thermochromatium (4.82%), and Geobacter (4.52%). This study highlights the potential of the integrated DCMFC–aerobic system, utilizing mixed actinomycetes strains, for the effective treatment of industrial dye effluents, offering both environmental and bioenergy benefits. Full article

Objectives: The aim of this study is to investigate the antimicrobial efficacy of antimicrobial photodynamic therapy (PDT) using natural photosensitizers (curcumin, riboflavin, and phycocyanin) and light-emitting diode (LED) irradiation against multispecies biofilms in an acrylic denture base model. Materials and Methods: Forty-five acrylic specimens were fabricated using heat-curing acrylic resin. The specimens were then infected with a mixed culture of bacterial and fungal species (including Streptococcus mutans, Streptococcus sanguinis, Candida albicans, and Candida glabrata) for 4 days. The acrylic discs were divided into nine groups, with each group containing five discs: control, 0.2% chlorhexidine, 5.25% sodium hypochlorite, curcumin, riboflavin, phycocyanin alone or along with LED. After treatment, the number of colony-forming units (CFUs) per milliliter was counted. In addition, the extent of biofilm degradation was assessed using the crystal violet staining method and scanning electron microscopy. Results: All experimental groups exhibited a significant reduction in colony numbers for both bacterial and fungal species compared to the control (p < 0.001). The PDT groups exhibited a statistically significant reduction in colony counts for both bacteria and fungi compared to the photosensitizer-only groups. Conclusions: The results of this in vitro study show that PDT with natural photosensitizers and LED devices can effectively reduce the viability and eradicate the biofilm of microorganisms responsible for causing denture infections. Full article

Candida albicans (C. albicans) and Streptococcus mutans (S. mutans) are frequently detected in the plaque biofilms of children with early childhood caries. This study investigated the effects of sucrose and farnesol on biofilm formation by the oral pathogens S. mutans and C. albicans, including their synergistic interactions. Biofilm formation dynamics were monitored using the Cell Index (CI). The CI for S. mutans increased in the brain–heart infusion medium, peaking at 10 h; however, the addition of sucrose reduced the CI. For C. albicans yeast cells, the CI increased at sucrose concentrations > 0.5%, peaking at 2 h. Mixed cultures of S. mutans and C. albicans yeast cells showed significantly higher CI values in the presence of sucrose, suggesting a synergistic effect on biofilm formation. Farnesol consistently suppressed biofilm formation by C. albicans yeast cells, even in the presence of sucrose, and higher farnesol concentrations resulted in greater inhibition. Regarding C. albicans hyphal cells, sucrose did not enhance biofilm formation, whereas farnesol significantly reduced biofilm formation at all concentrations tested. These findings elucidate the complex roles of sucrose and farnesol in biofilm formation by S. mutans and C. albicans and emphasize the potential of farnesol as an effective oral biofilm inhibitor. Full article

This study evaluates the suitability of three lactic acid bacteria (LAB) strains—Lactiplantibacillus plantarum, Lactobacillus acidophilus, and Apilactobacillus kunkeei—for use as probiotics in apiculture. Given the decline in bee populations due to pathogens and environmental stressors, sustainable alternatives to conventional treatments are necessary. This study aimed to assess the potential of these LAB strains in a probiotic formulation for bees through various in vitro tests, including co-culture interactions, biofilm formation, auto-aggregation, antioxidant activity, antimicrobial activity, antibiotic susceptibility, and resistance to high osmotic concentrations. This study aimed to assess both the individual effects of the strains and their combined effects, referred to as the LAB mix. Results indicated no mutual antagonistic activity among the LAB strains, demonstrating their compatibility with multi-strain probiotic formulations. The LAB strains showed significant survival rates under high osmotic stress and simulated gastrointestinal conditions. The LAB mix displayed enhanced biofilm formation, antioxidant activity, and antimicrobial efficacy against different bacterial strains. These findings suggest that a probiotic formulation containing these LAB strains could be used for a probiotic formulation, offering a promising approach to mitigating the negative effects of pathogens. Future research should focus on in vivo studies to validate the efficacy of these probiotic bacteria in improving bee health. Full article

(1) Background: In hospitals, medical and dental clinics, antiseptics or disinfectants play an essential role in the control of nosocomial infections. This study aimed to evaluate R. officinalis and P. paniculata glycolic extracts regarding: (I) their antimicrobial action on planktonic and biofilm (monotypic and cutaneous biofilm model—S. aureus, S. epidermidis and C. acnes); and (II) their cytotoxicity on human keratinocytes (HaCaT). (2) Methods: Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were performed (CLSI protocol M7-A6 and M11-A8). MTT analysis was used to evaluate the antibiofilm activity of the extracts on biofilms and their cytotoxicity on human keratinocytes. (3) Results: The combined glycolic extracts MIX A (75% P. paniculata + 25% R. officinalis); MIX B (50% P. paniculata + 50% R. officinalis); and MIX C (25% P. paniculata + 75% R. officinalis) promoted MBC values by 50 mg/mL on S. aureus, absent on S. epidermidis, and ranged 6.25–50 mg/mL for C. acnes. The cutaneous biofilm model was reduced more than 90%. In addition, it showed biocompatibility with human keratinocytes, resulting in percentages of viability greater than 50%. (4) Conclusions: The combination of extracts promoted antimicrobial action on planktonic cultures, and monotypic and heterotypic biofilms of skin pathogens. Additionally, these extracts are biocompatible against human keratinocytes. Full article

Campylobacter spp. are significant zoonotic agents, which cause annually millions of human cases of foodborne gastroenteritis worldwide. Their inclusion in biofilms on abiotic surfaces seems to play a pivotal role in their survival outside of the host, growth, and spread. To successfully mitigate the risks that arise with these bacteria, it is crucial to decrease their prevalence within the food production chain (from farm to the table), alongside the successful treatment of the resulting illness, known as campylobacteriosis. For this, the use of various antimicrobial agents remains actively in the foreground. A general-purpose biocide and cationic surfactant (benzalkonium chloride; BAC), a widely used macrolide antibiotic (erythromycin; ERY), and a naturally occurring organic acid (L(+)-lactic acid; LA) were comparatively evaluated in this work for their potential to inhibit both the planktonic and biofilm growth of 12 selected Campylobacter spp. (of which, seven were C. jejuni and five were C. coli) raw chicken meat isolates, all grown in vitro as monocultures. The inhibitory action of LA was also studied against four mixed-culture Campylobacter biofilms (each composed of three different isolates). The results showed that the individual effectiveness of the agents varied significantly depending on the isolate, growth mode (planktonic, biofilm), intercellular interactions (monocultures, mixed cultures), and the growth medium used (with special focus on blood presence). Thus, BAC exhibited minimum inhibitory concentrations (MICs), minimum bactericidal concentrations (MBCs), and minimum biofilm inhibitory concentrations (MBICs) that ranged from 0.5 to 16 μg/mL. Interestingly enough, these values varied widely from 0.25 to 1024 μg/mL for ERY. Concerning LA, the MICs, MBCs, and MBICs varied from 1024 to 4096 μg/mL, with mixed-culture biofilm formation always being more difficult to suppress when compared to biofilm monocultures. In addition, it was evident that intercellular interactions encountered within mixed-culture Campylobacter biofilms significantly influenced both the population dynamics and the tolerance of each consortium member to acid exposure. Overall, the findings of this study provide useful information on the comparative effectiveness of three well-known antimicrobial agents for the control of Campylobacter spp. under various growth modes (i.e., planktonic, biofilm, monocultures, mixed cultures) that could potentially be encountered in food production and clinical settings. Full article

Introduction: The purpose of this study was to assess the antifungal activity of silver nanoparticles (AgNPs) in combination with calcium hydroxide (Ca(OH)₂) against Candida albicans (C. albicans). Methods: AgNPs was mixed with pure Ca(OH)₂ powder in an aqueous base. A standard suspension (1 × 10⁸ bacterial cells/mL) of C. albicans was prepared in a 96-well plate and incubated on shaker at 37 °C in 100% humidity to allow fungal biofilm formation in infected dentin slices (n = 98). The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of AgNPs alone or with Ca(OH)₂ were determined. The samples were separately placed in 24-well tissue culture plates and divided into three experimental groups (0.03, 0.04, and 0.06) and three control groups; negative (saline) and positive chlorhexidine gel and Ca(OH)₂. Quantitative measurements of fungal activity by XTT colorimetric assay and qualitative measurements using confocal laser microscopy and scanning electron microscopy were performed. Results: The cell viability of C. albicans in the experimental groups was significantly reduced compared to the negative control group. The combination of (AgNPs (0.04%) and Ca(OH)₂) was the most potent against C. albicans. Conclusions: The findings demonstrated that combining silver nanoparticles with Ca(OH)₂ was more effective against C. albicans biofilm compared to Ca(OH)₂ alone, suggesting a combing effect. Full article

Dermatophytes associated with bacteria can lead to severe, difficult-to-treat infections and contribute to chronic infections. Trichophyton rubrum, Staphylococcus aureus, and Staphylococcus epidermidis can form biofilms influenced by nutrient availability. This study investigated biofilm formation by these species by utilizing diverse culture media and different time points. These biofilms were studied through scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), biomass, metabolic activity, and colony-forming units (CFUs). The results revealed that mixed biofilms exhibited high biomass and metabolic activity when cultivated in the brain heart infusion (BHI) medium. Both bacterial species formed mature biofilms with T. rubrum within 72 h, irrespective of media. The timing of bacterial inoculation was pivotal in influencing biomass and metabolic activity. T. rubrum’s development within mixed biofilms depended on bacterial addition timing, while pre-adhesion influenced fungal growth. Bacterial communities prevailed initially, while fungi dominated later in the mixed biofilms. CLSM revealed 363 μm thick T. rubrum biofilms with septate, well-developed hyphae; S. aureus (177 μm) and S. epidermidis (178 μm) biofilms showed primarily cocci. Mixed biofilms matched T. rubrum’s thickness when associated with S. epidermidis (369 μm), with few hyphae initially. Understanding T. rubrum and Staphylococcal interactions in biofilms advances antimicrobial resistance and disease progression knowledge. Full article

Antimicrobial photodynamic treatment (aPDT) with visible light plus water-filtered infrared-A irradiation (VIS-wIRA) and natural single- or multi-component photosensitizers (PSs) was shown to have potent antimicrobial activity. The aim of this study was to obtain information on the antimicrobial effects of aPDT-VIS-wIRA with lingonberry extract (LE) against bacteria that play a role in oral health. Planktonic bacterial cultures of the Gram-positive E. faecalis T9, S. mutans DSM20523, S. oralis ATCC 35037 and S. sobrinus PSM 203513, the Gram-negative N. oralis 14F2 FG-15-7B, F. nucleatum ATCC 25586, and V. parvula DSM, the anaerobic F. nucleatum ATCC 25586 and V. parvula DSM 2008, and the total mixed bacteria from pooled saliva and supra- and subgingival plaques of volunteers were all treated and compared. aPDT-VIS-wIRA with LE as PS significantly (p < 0.008) reduced the growth of all tested Gram-positive, Gram-negative, as well as aerobic and anaerobic bacterial strains, whereas without irradiation no reductions were seen (p < 0.0001). NaCl, with or without irradiation, was ineffective. After treatment with CHX 0.2%, the highest killing rate (100%) was observed, and no bacteria (0 log10 CFU) were cultivable. The method also significantly reduced all of the bacteria present in saliva and in the gingival biofilms. Three-dimensional visualization of viable and non-viable microorganisms revealed that LE penetrated deeper into the cell wall layers than CHX 0.2%. LE was an appropriate PS for eradicating microorganisms with VIS-wIRA, either in their planktonic form or in saliva and gingival plaque biofilms. These results encourage further investigation in order to determine which LE compounds contribute to the photosensitizing effect and to evaluate the size of the effect on maintaining oral health. Full article