Candida Biofilms Research Articles

Biofilm profile of Candida isolated from clinical specimens at a Tertiary Care hospital in India

Introduction Candida species are a leading cause of infections in immunocompromised hosts. Usage of indwelling medical devices provides ample opportunity for Candida biofilms to set up a nidus for infection that is not easily amenable to conventional antifungal therapy. Materials and Methods A total of 100 Candida isolates from patients with suspected candidiasis were tested for production of biofilm. Based on clinical history, 62% of the patients were found to have clinically significant infection with Candida while in 38% of the patients, Candida isolates represented commensals. Biofilm production was detected and graded by visual (test tube) and spectrophotometric (microtiter plate) methods. Results Fifty five percent of the Candida isolates produced biofilm. Biofilm producing abilities of clinically significant isolates (80%) was found to be significantly higher than commensals (20%). Biofilm positive Candida isolates were most commonly obtained from blood (34.5%). Biofilm production in non-albicans Candida (67.9%) was found to be significantly higher than C. albicans (38.6%). Majority of the biofilm positive Candida isolates produced Grade 2 (moderate) biofilm. Candida tropicalis accounted for maximum biofilm production comprising 20% Grade 4, 53.8% Grade 3 and 50% Grade 2 biofilm. Concordance in grading between the two methods was observed in 72.7% of the isolates. Spectrophotometric method was found to be more sensitive than visual method for detection of Candida biofilm. Conclusion The importance of studying Candida biofilms is to ascertain new therapeutics and techniques to manage these infections clinically and improve the outcome as these are associated with high morbidity, mortality and resistance to antifungal drugs.

Investigating the microbial composition of Recurrent Vulvovaginal Candidiasis samples and biofilm formation of Candida clinical isolates

Recurrent vulvovaginal candidiasis (RVVC) is a chronic and debilitating condition that is estimated to affect 138 million women annually. In spite of this high prevalence and its associated economic burden, pathogenesis of disease is poorly understood. The biofilm-forming yeast Candida albicans is reported as the causative pathogen in up to 90% of VVC cases and around 50% in RVVC disease. Despite the identification of Candidabiofilms on the vaginal mucosa, their associated therapeutic challenge in RVVC is still disputed. A panel of 100 HVS and cervico-vaginal lavage (CVL) samples were obtained for this trial. Patient questionnaires collected data on patient’s vaginal thrush history, treatments and any contraception currently used. Microbiological screening of clinical samples was performed, and microbiome analysis was utilized to determine predominant microbes present in each cohort. Non-C. albicans species (NCAS) such as intrinsically azole-resistant C. glabrata were found to account for 27% of RVVC cases, notably higher than historically reported. Additionally, microbiome analysis showed a reduction in Lactobacillus species associated with a healthy microbiome in RVVC compared to health. This study supports a previous clinical trial by our group using 300 HVS to investigate the epidemiology of RVVC showing an increasing prevalence of NCAS responsible for disease. Additionally, this work strengthens the hypothesis that formation of Candida biofilms on the vaginal mucosa could negatively impact clinical treatment. Further research to identify triggers for RVVC, and the pathogenesis of the predominant microbes involved, could considerably improve prevention and treatment options for women with recurrent, azole-resistant infections.

Antifungal and Antivirulence Activities of Hydroalcoholic Extract and Fractions of Platonia insignis Leaves against Vaginal Isolates of Candida Species.

Vulvovaginal candidiasis is a common fungal infection in women. In this study, Platonia insignis hydroalcoholic extract (PiHE) and its fractions were evaluated for antifungal and antivirulence activities against vaginal Candida species. Dichloromethane (DCMF) and ethyl acetate fractions (EAF) obtained from PiHE effectively inhibited the pathogen. Electrospray ionization mass spectrometry was used for identifying the main compounds in extracts. Minimal inhibitory and fungicidal concentrations (MIC and MFC, respectively) were determined by a broth microdilution assay. Furthermore, we evaluated the effect of the extract and fractions on the virulence properties of Candida albicans, and their cytotoxicity effect was determined on RAW 264.7 cells. Compounds found in extracts were flavonoid glycosides, mainly derivatives of quercetin and myricetin. Extracts showed antifungal potential, with the lowest MIC value for EAF (1.3 mg/mL) and inhibited Candida adherence and biofilm formation. EAF disrupted 48 h biofilms with an inhibition rate of more than 90%. The extract and its fractions exhibited no cytotoxicity. The antifungal effects were attributed to the ability of these extracts to alter the mitochondrial membrane potential for the release of pro-apoptotic factors in the cytosol. In conclusion, our data suggest that PiHE and EAF could act as novel candidates for the development of new therapeutic treatments against fungal infections.

Candida biofilm production is associated with higher mortality in patients with candidaemia

Candidaemia is a common life-threatening disease among hospitalised patients, but the effect of the Candida biofilm-forming ability on the clinical outcome remains controversial. The aim was to determine the impact of biofilms, specifically focusing on biofilm mass and metabolic activity, on the mortality in candidaemia. The clinical data of patients (n=127) treated at the University of Debrecen, Clinical Centre, between January 2013 and December 2018, were investigated retrospectively. Biofilm formation was assessed using the crystal violet and XTT assays, measuring the biofilm mass and metabolic activity, respectively. Isolates were classified as low, intermediate and high biofilm producers both regarding biofilm mass and metabolic activity. The susceptibility of one-day-old biofilms to fluconazole, amphotericin B, anidulafungin, caspofungin and micafungin was evaluated and compared to planktonic susceptibility. Intermediate/high biofilm mass was associated with significantly higher mortality (61%). All Candida tropicalis, Candida parapsilosis and Candida glabrata isolates originating from fatal infections were intermediate/high biofilm producers, whereas this ratio was 85% for Candida albicans. Solid malignancy was associated with intermediate/high biofilm producers (p=0.043). The mortality was significantly higher in infections caused by Candida strains producing biofilms with intermediate/high metabolic activity (62% vs. 33%, p=0.010). The ratio of concomitant bacteraemia was higher for isolates forming biofilms with low metabolic activity (53% vs. 28%, p=0.015). This study provides evidence that the Candida biofilms especially with intermediate/high metabolic activity are related to higher mortality in candidemia.

Cytotoxic Action of Artemisinin and Scopoletin on Planktonic Forms and on Biofilms of Candida Species.

We investigated the antifungal activities of purified plant metabolites artemisinin (Ar) and scopoletin (Sc) including inhibition, effects on metabolic activities, viability, and oxidative stress on planktonic forms and on preformed biofilms of seven Candida species. The characteristic minimum inhibitory concentration (MIC90) of Ar and Sc against Candida species ranged from 21.83–142.1 µg/mL and 67.22–119.4 µg/mL, respectively. Drug concentrations causing ≈10% CFU decrease within 60 min of treatments were also determined (minimum effective concentration, MEC10) using 100-fold higher CFUs than in the case of MIC90 studies. Cytotoxic effects on planktonic and on mature biofilms of Candida species at MEC10 concentrations were further evaluated with fluorescent live/dead discrimination techniques. Candida glabrata, Candida guilliermondii, and Candida parapsilosis were the species most sensitive to Ar and Sc. Ar and Sc were also found to promote the accumulation of intracellular reactive oxygen species (ROS) by increasing oxidative stress at their respective MEC10 concentrations against the tested planktonic Candida species. Ar and Sc possess dose-dependent antifungal action but the underlying mechanism type (fungistatic and fungicidal) is not clear yet. Our data suggest that Ar and Sc found in herbal plants might have potential usage in the fight against Candida biofilms.

Inhibition of Candida auris Biofilm Formation on Medical and Environmental Surfaces by Silver Nanoparticles.

Candida auris is an emerging pathogenic fungus implicated in healthcare-associated outbreaks and causes bloodstream infections associated with high mortality rates. Biofilm formation represents one of the major pathogenetic traits associated with this microorganism. Unlike most other Candida species, C. auris has the ability to survive for weeks on different surfaces. Therefore, there is an urgent need to develop new effective control strategies to combat the threat of C. auris. Advances in nanotechnologies have emerged that carry significant potential impact against Candida biofilms. We obtained pure round silver nanoparticles (AgNPs) (1 to 3 nm in diameter) using a microwave-assisted synthetic approach. When tested against C. auris, our results indicated a potent inhibitory activity both on biofilm formation (half maximal inhibitory concentration (IC50) of 0.06 ppm) and against preformed biofilms (IC50 of 0.48 ppm). Scanning electron microscopy images of AgNP-treated biofilms showed cell wall damage mostly by disruption and distortion of the outer surface of the fungal cell wall. In subsequent experiments AgNPs were used to functionalize medical and environmental surfaces. Silicone elastomers functionalized with AgNPs demonstrated biofilm inhibition (>50%) at relatively low concentrations (2.3 to 0.28 ppm). Bandage dressings loaded with AgNPs inhibited growth of C. auris biofilms by more than 80% (2.3 to 0.017 ppm). Also, to demonstrate long-lasting protection, dressings loaded with AgNPs (0.036 ppm) were washed thoroughly with phosphate-buffered saline, maintaining protection against the C. auris growth from cycles 1 to 3 (>80% inhibition) and from cycles 4 to 6 (>50% inhibition). Our results demonstrate the dose-dependent activity of AgNPs against biofilms formed by C. auris on both medical (silicone elastomer) and environmental (bandage fibers) surfaces. The AgNPs-functionalized fibers retain the fungicidal effect even after repeated thorough washes. Overall these results point to the utility of silver nanoparticles to prevent and control infections caused by this emerging pathogenic fungus.

Oxadiazolylthiazoles as novel and selective antifungal agents

Studying the structure-activity relationships (SAR) of oxadiazolylthiazole antibiotics unexpectedly led us to identify ethylenediamine- and propylenediamine-analogs as potential antimycotic novel lead structures. Replacement of the ethylenediamine moiety for the lead compound 7 with cis-diaminocyclohexyl group (compound 18) significantly enhanced the antifungal activity. In addition to the high safety margin of 18 against mammalian cells, it showed highly selective broad-spectrum activity against fungal cells without inhibiting the human normal microbiota. The antifungal activity of 18 was investigated against 20 drug-resistant clinically important fungi, including Candida species, Cryptococcus, and Aspergillus fumigatus strains. In addition to the low MIC values that mostly ranged between 0.125 and 2.0 μg/mL, compound 18 outperformed fluconazole in disrupting mature Candida biofilm.

In vitro Antifungal Effects of Berberine Against Candida spp. In Planktonic and Biofilm Conditions.

PurposeAntifungal resistance associated with the extensive use of antifungals and biofilm formation presents major clinical challenges. Thus, new therapeutic strategies for fungal infections are urgently required. This study aimed to evaluate the in vitro antifungal effects of the natural bioactive alkaloid berberine against Candida spp. in planktonic and biofilm conditions.MethodsUsing the CLSI M27-A3 reference method for broth dilution antifungal susceptibility testing of yeasts, the MICs for five standard strains comprised of Candida albicans (ATCC 10231, ATCC 90028), Candida krusei (ATCC 6258), Candida glabrata (ATCC 90030), Candida dubliniensis (MYA 646), and six clinical isolates (CLC1–CLC6) were tested. The 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay was used to evaluate the inhibitory effects of berberine against Candida biofilms. The optical density value at 490 nm was measured and illustrated using concentration-absorbance curves. Finally, the effects were quantified by confocal laser scanning microscopy (CLSM), and 3-dimensional reconstruction was performed. The viability inhibition rates, biofilm formation, and thickness decrease rates were tested and analyzed using independent-samples t-test. The differences among the five Candida strains were analyzed using one way ANOVA.ResultsThe MICs for the five standard strains described above were 80, 160, 10, 20, and 40 μg/mL, respectively, which was similar to that of the clinical isolates, suggesting the stable, broad-spectrum antifungal activity of berberine. Berberine exerted concentration-dependent inhibitory effects against Candida biofilms, which were enhanced with the maturation of Candida biofilms. Berberine decreased the viability of Candida biofilms, with inhibition rates by CLSM ranging from 19.89 ± 0.57% to 96.93 ± 1.37%. Following 3-dimensional reconstruction, the biofilms of the berberine-treated group displayed a poorly developed architecture, and the biofilm thickness decrease rates ranged from 15.49 ± 8.45% to 30.30 ± 15.48%.ConclusionBerberine exhibited significant antifungal activity in Candida spp. The results provide a useful reference for multiple Candida infections and biofilm infections associated with antifungal resistance. Therefore, berberine might have novel therapeutic potential as an antifungal agent or a major active component of antifungal drugs.

Discovery of a novel and selective fungicide that targets fungal cell wall to treat dermatomycoses: 1,3-bis(3,4-dichlorophenoxy)propan-2-aminium chloride.

Fungal infections are highly prevalent and are responsible for high rates of morbidity and mortality. In this context, the search for new treatment alternatives is very relevant. Analyse chemical compounds for antifungal potential against dermatomycosis fungi. The antifungal activity of 121 compounds, intermediates or derivatives of 1,3-bis(aryloxy)propane substituted at C-2 (111 compounds) and isothiouronium derivatives (10 compounds) was investigated through susceptibility tests, mechanism of action, toxicity and hydrogel incorporation. The compound 1,3-bis(3,4-dichlorophenoxy)propan-2-aminium chloride (2j) was the most active fungicide against dermatophytes and Candida spp., at very low concentrations (0.39-3.12µg/mL), including action on resistant and multidrug-resistant clinical strains. Compound 2j has presented a promising toxicity profile, showing selectivity index >10, relative to human lymphocytes. The compound was classified as non-irritant by the HET-CAM test and did not cause histopathological alterations in pig ear skin, thus presenting an excellent perspective for topical application. 2j targets the fungal cell wall, which was confirmed by scanning electron microscopy, which also indicated the additional ability of 2j to inhibit the Candida albicans pseudohyphae formation and biofilm of Microsporum canis. Compound 2j was incorporated in a hydrogel with bioadhesive potential. The results of the human skin permeation showed that 2j remained significantly in the epidermis, ideally for the dermatomycosis treatment. Therefore, the compound 2j demonstrated the potential for antifungal drug development, with a action mechanism elucidated and already applied in a semisolid formulation as a new therapeutic option for fungal skin infections.

Inhibition-Disruption of Candida glabrata Biofilms: Symmetrical Selenoesters as Potential Anti-Biofilm Agents.

Candida glabrata is one of the most prevalent pathogenic Candida species in dental plaque on tooth surfaces. Candida biofilms exhibit an enhanced resistance against most antifungal agents. Thus, the development of alternative more potent and effective antimicrobials is required to overcome this resistance. In this study, three novel fluorinated derivatives and nine selenoester compounds were screened as novel antifungal and antibiofilm agents against C. krusei, C. parapsilosis, and C. glabrata (N = 81 dental isolates). C. glabrata strains were susceptible only to fluorinated compounds while C. krusei, C. parapsilosis, and C. glabrata were susceptible to the action of the selenoesters. The evaluated symmetrical selenoester compounds presented very good antifungal activity against all the tested C. glabrata dental isolates (1–4 μg/mL of minimum inhibitory concentration-MIC). The most active compound (Se-5) was able to inhibit and disperse C. glabrata biofilms. These results demonstrated that selenoesters may be novel and promising biocide agents against C. glabrata clinical dental isolates.

In Vitro and In Vivo Anti-Candida spp. Activity of Plant-Derived Products.

Candidiasis therapy, especially for candidiasis caused by Candida non-albicans species, is limited by the relatively reduced number of antifungal drugs and the emergence of antifungal tolerance. This study evaluates the anticandidal activity of 41 plant-derived products against Candida species, in both planktonic and biofilm cells. This study also evaluates the toxicity and the therapeutic action of the most active compounds by using the Caenorhabditis elegans–Candida model. The planktonic cells were cultured with various concentrations of the tested agents. The Cupressus sempervirens, Citrus limon, and Litsea cubeba essential oils as well as gallic acid were the most active anticandidal compounds. Candida cell re-growth after treatment with these agents for 48 h demonstrated that the L. cubeba essential oil and gallic acid displayed fungistatic activity, whereas the C. limon and C. sempervirens essential oils exhibited fungicidal activity. The C. sempervirens essential oil was not toxic and increased the survival of C. elegans worms infected with C. glabrata or C. orthopsilosis. All the plant-derived products assayed at 250 µg/mL affected C. krusei biofilms. The tested plant-derived products proved to be potential therapeutic agents against Candida, especially Candida non-albicans species, and should be considered when developing new anticandidal agents.

RAFT-Derived Polymethacrylates as a Superior Treatment for Recurrent Vulvovaginal Candidiasis by Targeting Biotic Biofilms and Persister Cells.

BackgroundVulvovaginal candidiasis (VVC) is a common infection in need of more effective treatment. Formation of epithelium-associated Candida biofilms and the presence of persister cells are among the major contributing factors to the recurrence of this condition. We have previously developed RAFT-derived polymethacrylates that are effective in killing C. albicans biofilms in vitro. This study aimed to examine the clinical potential of polymethacrylates as antifungals for treatment of recurrent VVC (RVVC).MethodsA mouse model of VVC was used to establish vaginal epithelium-associated biofilms, using C. albicans isolates from VVC/RVVC patients. A comparison was made of the efficacies of polymethacrylates and conventional antifungals, clotrimazole and nystatin, in killing Candida in epithelium-associated biofilms in vivo. Ex vivo biofilms were used for Candida population profiling and to quantify persister cells in vaginal epithelia. The potency of polymethacrylates and conventional antifungals against persister cells, either as sole agents or in combination, was assessed.ResultsPolymethacrylates showed negligible local toxicity, resistance to vaginal acidity, and outstanding in vivo activity against vaginal epithelium-associated C. albicans biofilms. In vivo tests polymethacrylates outperformed the conventional antifungals, nystatin and clotrimazole at concentrations 50 times below the over-the-counter concentrations. Using polymethacrylates was associated with fewer persister cells, and better eradication of persister cells pre-selected by conventional antifungals.ConclusionThis study systematically assessed the clinical potential of RAFT-derived polymethacrylates as an effective treatment for VVC/RVVC in a mouse model. Polymethacrylates effectively killed vaginal epithelium-related C. albicans in vivo by specially targeting biotic biofilms and persister cells. Treatment presented negligible local toxicity.

Inhibitory Effect of 5-Aminoimidazole-4-Carbohydrazonamides Derivatives Against Candida spp. Biofilm on Nanohydroxyapatite Substrate.

Candida can adhere and form biofilm on biomaterials commonly used in medical devices which is a key attribute that enhances its ability to cause infections in humans. Furthermore, biomaterial-related infections represent a major therapeutic challenge since Candida biofilms are implicated in antifungal therapies failure. The goals of the present work were to investigate the effect of three 5-aminoimidazole-4-carbohydrazonamides, namely (Z)-5-amino-1-methyl-N'-aryl-1H-imidazole-4-carbohydrazonamides [aryl = phenyl (1a), 4-fluorophenyl (1b), 3-fluorophenyl (1c)], on Candida albicans and Candida krusei biofilm on nanohydroxyapatite substrate, a well-known bioactive ceramic material. To address these goals, both quantitative methods (by cultivable cell numbers) and qualitative evaluation (by scanning electron microscopy) were used. Compounds cytocompatibility towards osteoblast-like cells was also evaluated after 24h of exposure, through resazurin assay. The three tested compounds displayed a strong inhibitory effect on biofilm development of both Candida species as potent in vitro activity against C. albicans sessile cells. Regarding cytocompatibility, a concentration-dependent effect was observed. Together, these findings indicated that the potent activity of imidazole derivatives on Candida spp. biofilms on nanohydroxyapatite substrate, in particular compound 1c, is worth further investigating.

Biofilm inhibiting properties of compounds from the leaves of Warburgia ugandensis Sprague subsp ugandensis against Candida and staphylococcal biofilms.

Warburgia ugandensis Sprague subspecies ugandensis is a plant widely distributed in Eastern, Central and Southern Africa. In humans, it is used to treat respiratory infections, tooth aches, malaria, skin infections, venereal diseases, diarrhea, fevers and aches. This study aims to identify the bioactive compounds against clinically important biofilm-forming strains of Candida and staphylococci that are responsible for tissue and implanted device-related infections. Using a bioassay-guided fractionation approach, hexane -, ethanol -, acetone - and water extracts from the leaves of W. ugandensis, their subsequent fractions and isolated compounds were tested against both developing and preformed 24 h-biofilms of Candida albicans SC5314, Candida glabrata BG2, Candida glabrata ATCC 2001, Staphylococcus epidermidis 1457 and Staphylococcus aureus USA 300 using microtiter susceptibility tests. Planktonic cells were also tested in parallel for comparison purposes. Confocal scanning laser microscopy was also used to visualize effects of isolated compounds on biofilm formation. Warburganal, polygodial and alpha-linolenic acid (ALA) were the major bioactive compounds isolated from the acetone extract of W. ugandensis. For both warburganal and polygodial, the biofilm inhibitory concentration that inhibits 50% of C. albicans developing biofilms (BIC50) was 4.5 ± 1 and 10.8 ± 5 μg/mL respectively. Against S. aureus developing biofilms, this value was 37.9 ± 8 μg/mL and 25 μg/mL with warburganal and ALA respectively. Eradication of preformed 24 h biofilms was also observed. Interestingly, synergy between the sesquiterpenoids and azoles against developing C. albicans biofilms resulted in an approximately ten-fold decrease of the effective concentration required to completely inhibit growth of the biofilms by individual compounds. The hydroxyl group in position C-9 in warburganal was identified as essential for activity against staphylococcal biofilms. We also identified additional promising bioactive sesquiterpenoids; drimenol and drimendiol from the structure-activity relationship (SAR) studies. ALA and four sesquiterpenoids: polygodial, warburganal, drimenol and drimendiol, have shown biofilm-inhibitory activity that has not been reported before and is worth following up. These compounds are potential drug candidates to manage biofilm-based infections, possibly in combination with azoles.

Synthesis of new pyrazolo[5,1-c][1,2,4]triazines with antifungal and antibiofilm activities

Fungal infection is a significant global health challenge in part due to the emergence of strains exhibiting resistance to nearly all classes of antifungals. This underscores the urgent need for the development of new antifungal agents that can circumvent this burgeoning problem. For the present research, a new selected set of pyrazolo[5,1-c][1,2,4]triazine derivatives 3a–g was prepared in high yield via the reaction of N1-(5-methylpyrazol-3-yl)hydrazonoyl chloride 1 with morpholine, thiomorpholine, 4-phenylpiperidine and N-(substituted)piperazines. The new compounds were evaluated for their in vitro antifungal and antibacterial activities. The screening revealed compounds with specific activity against pathogenic fungi, including Candida albicans, Candida auris, and Cryptococcus. Compound 3d, which incorporated N-phenylpiperazine moiety, exhibited the highest growth inhibition against C. albicans with a minimum inhibitory concentration of 16 µg/mL. The compounds were superior to fluconazole in inhibiting Candida biofilm mass at sub-inhibitory concentration. Furthermore, the MTS assay confirmed that compounds 1 and 3d exhibited an excellent toxicity profile (not toxic, up to 256 μg/mL, for mammalian cells). Collectively, the presented results demonstrate that the synthesized pyrazolo-triazines warrant further exploration for potential use as antifungal agents.

Marine bacterial DNase curtails virulence and disrupts biofilms of Candida albicans and non-albicans Candida species

Candida is one of the most prevalent fungal pathogens in clinical settings which form antibiotic-resistant biofilms on biomedical devices. Hence, there is a need for non-antimicrobial alternatives to combat these infections. The present study investigates the anti-biofilm effect of marine bacterial DNase by targeting the eDNA present in the biofilms of Candida spp. A strain of Vibrio alginolyticus (AMSII) which showed enhanced DNase activity was isolated from marine sediment. Treatment of young and mature Candida biofilms with purified marine bacterial DNase (MBD) caused a 60-80% reduction in biofilm biomass, similar to treatment with DNase I from Bovine pancreas. Scanning electron microscopy showed that MBD significantly reduced the formation of biofilms on urinary catheters and more importantly prevented the virulent yeast to hyphae dimorphic switch in C. albicans. The present study identified a potential non-antibiotic alternative therapy to eradicate Candida biofilms and can be used to develop enzyme fabricated antifouling indwelling medical devices.

Use of scanning electron microscopy to study Candida albicans biofilms on the surface of base plastics of removable orthopedic constructions.

The most common pathology in the clinic of orthopedic dentistry is the presence of partial adentia in patients, manifested in the form of defects of dentition of various localization and length. Removable orthopedic structures in the oral cavity are a potential place for adhesion and colonization of microorganisms. The aim of the research was to study Candida albicans biofilms on the surface of base plastics of removable orthopedic structures using scanning electron microscopy. 175 cultures of C. albicans were isolated and identified from the oral mucosa of patients at various stages of orthopedic rehabilitation. When studying the surface of samples of plastics of hot and cold type polymerization and Candida biofilms using a JEOL JCM 5700 scanning electron microscope (JEOL, Japan), features of biofilm formation were established. An assessment of the nature of the manifestation of the hemagglutinating activity of clinical strains of Candida fungi in the hemagglutination test with human erythrocytes I (O), II (A) of the human and guinea pig blood groups was carried out. The total number of hemagglutinating strains was 37.14%, with the prevalence of the proportion of manna-resistant (MRHA) cultures - 23.43% of cases. Micrographs of the C. albicans yeast-like biofilm biofilm were obtained on the surface of hot and cold-type plastics in incubation dynamics. Scanning electron microscopy revealed the most pronounced changes in the surface of hot plastics of polymerization compared to cold plastics with long incubation of C. albicans, which characterize the loosening of plastics and the appearance of cracks on the surface, and the cracking of a yeast-like fungus biofilm was noted.

Structural insights into conformation of amphiphilic quaternary ammonium chitosans to control fungicidal and anti-biofilm functions

Fungal biofilm formation is an emerging problem in a wide range of health-related applications. This study aims to design and synthesize amphiphilic quaternary ammonium chitosans (AQACs) that could bind onto fungal biofilms to kill adherent fungal cells, and establish their structural/fungicidal activity relationships. AQACs with different hydrophobic alkyl chain length (C4, C8, and C12) were synthesized by quaternization of 3-bromopropionic acid with the corresponding tertiary amines, followed by reacting with chitosan using the EDC/NHS chemistry. The new AQACs were soluble in water, yet formed self-aggregates in the solution with different sizes. In antifungal tests against free-floating Candida albicans, shorter alkyl chains (C4) in the AQACs resulted in the most potent fungicidal effect. However, in the treatment of Candida biofilms formed on solid surfaces, AQACs with longer alkyl chains (C8 and C12) were much more effective than their shorter chain counterpart (C4). The effects of alkyl chain self-aggregation on the opposite trend in fungicidal and anti-biofilm activities were discussed. All the AQACs showed excellent cytocompatibility with mammalian cells.

Impact of various finishing and polishing techniques and composite materials on Candida albicans biofilm formation.

Candida albicans biofilms are commonly associated with severe oral infections. In dentistry, prosthetic and restorative materials are potential structures for the adhesion of C. albicans facilitating the formation of Candida biofilm and infection. Three composite resins (Charisma Classic, Sonic Fill, Estelite ∑ Quick) and two finishing-polishing systems (Biscover LV, Dental Finishing Disc) were evaluated for Candida biofilm formation. A Candida biofilm assay showed that both the resin and the finishing/polishing procedures affect Candida biofilm formation. Specifically, Candida biofilm formation was significantly lower in Sonic Fill resin than both Charisma Classic and Estelite ∑ Quick (P = .021). The type of finishing and polishing procedure also significantly affected the Candida biofilm formation to composite material (P < .001). Candida biofilm formation was more advanced after Biscover LV procedure than Dental Finishing Disc procedure.

Invasive Fungal Infections in the Pediatric Intensive Care Unit.

Invasive Fungal Infections in the Pediatric Intensive Care Unit.