Antimicrobial photodynamic therapy (aPDT) is a promising alternative in the microorganisms inactivation and infections caused by pathogenic fungi in planktonic and biofilm forms. The lack of development of resistance against this therapy makes it indicated even for the treatment of fungi resistant to antifungals. There are numerous invitro studies that demonstrate the benefits of aPDT on pathogenic fungi with a high mortality rate, such as those of the genus Candida spp. However, studies show variations in the parameters used in their protocols, which makes it difficult to identify a more effective protocol for advancing studies in animal models. This review aimed to highlight the reported discrepancies between invitro studies of aPDT as an antifungal therapy, making the translational protocol for invivo studies a complex task.

Partial NaCl replacement with KCl enhances microbial dynamics and antioxidant potential in fermented Nabali Baladi olives

Fungal species of the genus Candida are commensal in immunocompetent individuals but may become pathogenic under immunosuppressive conditions. Likewise, Pichia kudriavzevii has emerged as an opportunistic yeast associated with nosocomial infections, especially in immunocompromised patients. The increasing resistance to conventional antifungal agents has stimulated the search for alternative therapies, including essential oils. This study investigated the chemical composition and antifungal activity of the essential oil from fresh leaves of Myrcia spectabilis (EOFLMS) against Candida spp. and P. kudriavzevii. The oil was obtained by hydrodistillation and analyzed by GC-MS, revealing a predominance of sesquiterpenes, mainly germacrene B, germacrene D, bicyclogermacrene, α-cadinol, cis-cadin-4-en-7-ol, and cubeban-11-ol. Antifungal activity was evaluated through fungal growth curves, IC50, Minimum fungicidal concentration (MFC), and pleomorphism assays. EOFLMS alone showed moderate activity, especially against Candida albicans, reducing fungal growth by 99.25% (IC50=1215 µg/mL). In combination with fluconazole, EOFLMS enhanced antifungal activity against C. albicans and Candida tropicalis, with IC50 values of 22.91 and 12.38 µg/mL, respectively. EOFLMS also inhibited hyphal and pseudohyphal formation, suggesting interference with fungal virulence factors.

Vulvovaginal candidiasis (VVC), also known as vaginal thrush, is an infection of the vulvovaginal mucosa caused by fungi of the Candida genus. Particularly for patients suffering from recurrent infection, the disease has a significant impact on their quality of life. The still unknown aspects of disease pathogenesis, as well as factors driving the development of infections and recurrence, represent a challenge for both clinical practitioners and patients. Mouse models and patient studies have suggested important roles of the microbiome, deployment of fungal pathogenicity mechanisms in the vagina, and dysregulated immune responses for VVC pathology. Dissecting their individual contributions can reveal specific processes associated with infection and may inspire novel therapeutic strategies. Epithelial in vitro infection models have been playing a key role in dissecting a crucial interaction during VVC, the invasion and infection of the vaginal mucosa. They have been instrumental in characterizing candidalysin as a fungal toxin that damages epithelial cells and elicits initial inflammatory responses to catalyze downstream inflammation. Moreover, they have also revealed potential protective immune pathways. Such a standardized epithelial cell infection model offers high versatility and compatibility with different downstream assays to link epithelial responses with other processes during VVC. This protocol describes a general A-431 vulvovaginal epithelial cell–Candida infection model in detail and provides several adaptations, such as live-cell imaging and mRNA silencing, as well as possible follow-up readouts, like the quantification of cytokine release, cytotoxicity, and neutrophil recruitment to study diverse processes relevant to VVC research.Key features• This protocol describes the use of the A-431 epidermoid carcinoma cell line as an in vitro infection model of vulvovaginal candidiasis.• The protocol can be adapted to answer research questions relevant to vulvovaginal candidiasis (epithelial damage, release of inflammatory cytokines, recruitment and activation of neutrophils).• Live-cell imaging can be used to study dynamic infection processes.• mRNA silencing can be applied to interrogate the function of genes of interest in the host–pathogen interaction.This protocol is used in:

Background: Candida tropicalis is a pathogenic yeast species responsible for infections within the Candida genus and is identified as the most virulent species after C. albicans, partly due to its ability to form biofilms. Objective: This study analyzes the antifungal efficacy of a newly synthesized dendron, BD132 dendron, against C. tropicalis. Results: The compound showed a strong antifungal activity with promising minimum inhibitory concentration (MIC) and minimum biofilm inhibitory concentration (MBIC) values. Combination therapy with AgNO3 and amphotericin B showed additive and synergistic effects, respectively, enhancing antifungal efficacy and potentially reducing cytotoxicity. The dendron did not alter key enzyme activities, and scanning electron microscopy revealed significant morphological alterations, including increased cell size and surface damage, indicating membrane disruption. In addition, the BD132 dendron did not induce resistance, and stability studies indicated a slight MIC decrease at 4 °C and −20 °C after 15 days, with stable minimum fungicidal concentration (MFC), suggesting potential for long-term use. Conclusions: These findings highlight the potential of this dendron in combination therapies to treat C. tropicalis infections.

Claiming 3.8 million lives annually, fungal pathogens represent a major health threat, where Candida species are the leading cause with >40% mortality rates. Despite rising prevalence of life-threatening fungal species/strains, progress in developing new antifungal agents remains limited. Here, we report a new synthetic membrane-active cationic supramolecule Gua-SMACS-16 that exhibits high antifungal potency and selectivity for Candida tropicalis (MIC = 0.4-0.8µM) without affecting other phylogenetically close species in the Candida genus, non-Candida fungal, or common opportunistic bacterial species in the human microbiome. Gua-SMACS-16 exerts its activity by disrupting the cytoplasmic membrane, while its C. tropicalis selectivity is attributed to the species' low cell wall β-glucan level, which fails to block the supramolecule entry. It also exhibits minimal toxicity to mammalian cells and zebrafish embryos, suggesting a high clinical translation potential. Moreover, an ultra-strong synergy was observed between Gua-SMACS-16 and caspofungin, a clinical antifungal drug inhibiting 1,3-β-glucan synthase, reducing their MICs by orders of magnitude against all tested Candida species including the intractable C. auris. Together, this work highlights the importance of cell wall glucan for membrane-active antifungal amphiphiles, provides a novel design principle for species-specific antifungal agents, and uncovers a new approach to developing synergizers for combination antifungal therapy.

Background. Urolithiasis is a chronic multifactorial disease characterized by the formation of renal calculi of various salt compositions. Determination of the composition of salts that form calculi is necessary for adequate treatment and prevention of the transition of the stone formation process to the active stage. However, the possibility of determining stone-forming salts in the early stages of the disease (before stone formation) remains a problem for clinical practice. Purpose of the study : to prove the possibility of determining stone-forming salts of oxalate-calcium composition by examining urine using the systemic self-organization method (Lithos-system technology) in patients at the early (preclinical) stage of nephrolithiasis, as well as at the early stage of disease recurrence. Material and methods. Method of systemic self-organization of a urine drop (Lithossystem technology), method of X-ray spectral microanalysis. Results . In urine facies (structured films of dehydrated drops) of 57 patients, the systemic self-organization of urine method revealed an active process of stone formation, as indicated by multiple crystalline formations in the marginal (protein) zone of the facies. Microscopy of these crystalline formations in polarized light showed that they were crystallized structures of the fungus of the genus Candida spp. It is known that the main product of the vital activity of Candida spp. is oxalic acid. Electron probe study (X-ray spectral microanalysis method) showed that the chemical composition of crystalline formations in the protein zone of the facies was represented by calcium oxalate. Thus, the simultaneous presence of crystalline formations and Candida spp. structures in the protein zone of urine facies indicates the formation of oxalate-calcium stones. Conclusion . The method of systemic self-organization of urine can be used to diagnose the process of stone formation based on calcium oxalate in urolithiasis at all (including preclinical) stages of disease development in clinical diagnostic laboratories.

Candidiasis, caused by yeasts of the Candida genus, is increasingly characterized by a high prevalence of clinical isolates resistant to conventional antifungals, rendering the development of novel therapeutic strategies paramount. Drug repurposing has emerged as a key strategy, utilizing established pharmaceuticals for indications beyond their original design; notably, haloperidol (HAL) has shown promising antimicrobial potential. In this context, the present study evaluates the activity of haloperidol, both as a monotherapy and in combination with conventional antifungals, against fluconazole-susceptible and fluconazole-resistant Candida spp. clinical strains. Furthermore, we investigate the underlying mechanisms of its antifungal action. Experimental approaches included broth microdilution assays to determine the Minimum Inhibitory Concentration (MIC), checkerboard assays for synergistic analysis, and cellular assessments via flow cytometry and fluorescence microscopy. Haloperidol displayed MIC values between 26.67 and 256 μg/mL. Synergistic interactions were identified between haloperidol and the azoles fluconazole and itraconazole, alongside a 2.5 % synergy rate with amphotericin B. Additionally, mechanistic assays confirmed that haloperidol induces programmed cell death (apoptosis) in C. albicans and C. auris strains. The oxidative stress caused by haloperidol altered Ca2+ homeostasis, followed by mitochondrial membrane depolarization, reduced ATP production, cytochrome c release into the cytosol and metacaspase activation, reduced viability, phosphatidylserine externalization, promoted fragmentation, damage and methylation of DNA. It also induced expression of genes related to oxidative stress. It reduced mitochondrial depolarization and decreased the reduction of glutathione (GSH), causing morphological alterations. The results suggest the apoptotic pathway as the main antifungal mechanism of haloperidol.

Candidemia, caused by the opportunistic Candidaspp., poses a severe global health threat, characterized by mortality rates reaching 75% and the increasing prevalence of multidrug-resistant biofilms. In this study, we targeted the Agglutinin-Like Sequence 3 (Als3) adhesin protein, the primary mediator of Candida species attachment, as a non-lethal strategy to inhibit biofilm formation. Using molecular docking, we screened a library of 1570 natural carotenoids against the conserved N-terminal domain of Als3 of Candida albicans and compared to five positive controls. Our virtual screening identified five compounds (mangicrocin, MODU, α-zeacarotene-3,17'-diol, BTTB, and (15Z)-zeaxanthin-3'-rhamnoside), with the best binding pose and the least binding energy. Among them, mangicrocin showed best binding affinity (docking score - 7.18kcal/mol, ΔGbind - 35.23kcal/mol) against C. albicans Als3, compared to the in vitro and in silico positive control, rutin (docking score - 5.772kcal/mol, ΔGbind - 30.59). It consistently achieved the best docking scores across C. albicans Als family proteins (Als1 and Als5) and Als3 proteins of other species in the Candida genus like C. auris, C. tropicalis, C. parapsilosis, and C. dubliniensis. Molecular electrostatic potential (MESP) mapping analysis further clarified that the hydrophobic polyene backbone and the negative electrostatic potential of the OH groups on the aromatic rings aid in anchorage to the key cadherin-binding residues-Lys59 and Ser170 in the Als3 binding pocket. Density Functional Theory (DFT) analysis shows high electrophilicity and a narrow energy gap contributes to mangicrocin's interactions and chemical reactivity. Molecular dynamics simulations suggested stable retention of mangicrocin within the Als3 binding pocket, supported by stable structural parameters and favorable interaction energy profiles. In silico ADMET profiling concludes these top five hits had limited toxicity and are druggable. As this study is entirely computational, these findings are predictive and require future in vitro validation to confirm biological efficacy.

ABSTRACT Objectives. To identify Candida species using MALDI-TOF and analyze their antifungal susceptibility from Candida genus isolates in hospitalized patients with COVID-19 between November 2020 and April 2022.Materials and methods. An observational, descriptive, and cross-sectional study based on the secondary analysis of microbiological and clinical-epidemiological data from 260 Candida isolates, primarily from urine, respiratory secretions, and blood cultures, referred to the National Reference Laboratory of Mycology of the National Institute of Health. Identification was performed using phenotypic techniques and MALDI-TOF, and antifungal susceptibility was evaluated using disk diffusion and broth microdilution according to Clinical and Laboratory Standards Institute (CLSI) criteria.Results.Candida albicans was the most prevalent species (64.6%), followed by Candida tropicalis, Candida glabrata, Candida parapsilosis, and Candida auris. Most isolates showed susceptibility to voriconazole and caspofungin, while varying percentages of susceptibility to fluconazole were observed, especially in C. glabrata and C. auris. Minimum inhibitory concentration (MIC) values showed variability between species and antifungals.Conclusions. Non-albicans species represented a considerable proportion of the analyzed isolates. These findings describe the pattern of species and antifungal susceptibility in strains referred to the National Reference Laboratory of Mycology of the National Institute of Health during the COVID-19 pandemic.

The research aimed to conduct a survey and statistical study in Najaf province in 2019 for both Candida and Monilia fungi, where it was observed that the Candida genus prevailed by 8.05%, while the percentage recorded for the Monilia genus was 4.06%. The study also included the rates of difference in infection between males and females, where the results showed that females were more likely to be infected than males, with 6.10% for females and 6.01% for males. The results also showed that Al-Manadhera Hospital is the most recorded cases of infection with these two fungi with 7.56%, while the lowest percentage was recorded in Al-Hakim General Hospital with 0.75%. As for the difference in infection during the months of the year, October recorded the highest infection rates with 1.40%, while January was the lowest with 0.75%.

Tracheoesophageal puncture (TEP) with voice prosthesis (VP) insertion is standard procedure to restore speech after total laryngectomy. Microbial colonization on VPs leads to various complications. The aim of this study was to evaluate the microbiome colonization in tracheoesophageal fistulas as potential factors reducing the longevity of VPs and to compare the impact of the care regimens for laryngectomized patients on the microbiome. A total of 52 patients who underwent laryngectomy with primary TEP and VP insertion were included and randomly assigned to two groups: the regular exchange arm (REA) and the leakage exchange arm (LEA). Two separate swabs, one from the trachea and the other one from the tracheoesophageal fistula, were collected at the time of VP replacement. Fistula swab cultures showed bacterial colonization in 98.1% of cases, with Staphylococcus aureus being the most commonly identified bacterium (51.9%). Yeast colonization was present in 40.4% of fistula swabs, with Candida species identified. Tracheal swabs yielded positive bacterial cultures in 92.3% samples and fungal in 19.2%. The most commonly cultured bacterium was Staphylococcus aureus (51.9%), and the most common fungus was the genus Candida (100%). None of the identified microbiological factors significantly differentiated patients based on the regularity of VP replacement (p = 0.603-1). The identified bacteria and fungi were similar regardless of complications, comorbidities or adjuvant treatment. Bacteria and yeast were identified in almost all swabs from tracheoesophageal fistula and trachea of laryngectomized patients with VPs. A frequent co-colonization of bacteria and fungi was observed, with Candida albicans and Staphylococcus aureus being the most common. Follow-up of patients with VP, including the regularity of VP replacements, should not be dependent on microbial colonization of the fistula or trachea.

Objectives: Topical corticosteroid therapy (TCST) is the primary treatment for oral lichen planus (OLP), but an increased risk of oral candidiasis remains a common adverse effect. This study aimed to identify microbial predictors of candidiasis risk by comparing baseline oral bacteriome and mycobiome profiles of OLP patients who developed candidiasis following TCST with those who did not. Methods: A nested case-control study was conducted among twelve participants from an ongoing randomized controlled trial. Baseline oral rinse samples were collected from symptomatic OLP patients prior to TCST—six who later developed candidiasis (based on clinical and microbiological criteria) at 2–4 weeks post-treatment and six matched controls who did not. Samples were cultured on Sabouraud-dextrose agar with antibiotics to quantify Candida colonization, and DNA was extracted for 16S rRNA and ITS sequencing (Illumina MiSeq) to profile bacterial and fungal communities. Bioinformatics analysis included relative abundance, α-diversity, β-diversity (UNIFRAC), and LEfSe analyses using QIIME2. Results: At baseline, 67% of patients who later developed candidiasis exhibited high Candida colonization compared to 33% in the control group. Both groups showed dominance of Streptococcus and Candida genera; however, the candidiasis group demonstrated significantly lower bacterial α-diversity (Shannon index, p = 0.041), while fungal α-diversity remained similar. Bacterial β-diversity (unweighted UNIFRAC, p = 0.013) differed significantly between groups, but not fungal profiles. LEfSe analysis identified enrichment of Streptococcaceae and several bacterial taxa in the candidiasis group, while distinct bacterial and fungal taxa were enriched among controls. Conclusions: OLP patients who developed oral candidiasis following TCST exhibited lower bacterial diversity and higher baseline Candida colonization before treatment. Specific bacterial taxa, including Streptococcus spp., may serve as potential microbial biomarkers for predicting candidiasis risk. These findings underscore the potential role of the oral microbiome in guiding personalized management strategies for OLP patients receiving corticosteroid therapy.

Yeast Spectrum and antifungal susceptibility in a Hospital in Southwestern China: A retrospective Study (2020-2024).

BLOODSTREAM INFECTION CAUSED BY SACCHAROMYCES CEREVISIAE: CASE REPORT

Candidiasis is a fungal disease affecting poultry, primarily caused by the genus Candida. A poultry desi bird of 40 days old was presented for necropsy to the Department of Veterinary Pathology, Veterinary College, Hassan with a history of weakness and anorexia. It was from a flock with a total of 1100 birds in which death of 10-20 birds each day were recorded. On external examination, body condition appeared poor, conjunctival mucous membrane was pale. The keel bone was prominent. Internal examination revealed intestine with moderate diffuse petechial haemorrhage and turkey towel appearance of crop. The sample from the crop was submitted to Department of Microbiology for culture and identification. The sample cultured on Sabouraud dextrose agar revealed thick creamy colonies typical of yeast. Microscopic examination of the colonies stained by Gram’s stain revealed budding yeast cells characteristic of Candida species. Antifungal assay performed using the pure colony of Candida spp. by disc diffusion method revealed that the isolate was sensitive to all the antifungal agents used.

Piper tuberculatum Jacq. essential oil and its impact on the control of Candida spp. and Pichia kudriavzevii: increased efficacy of fluconazole and inhibition of virulence.

Chronic gastroduodenitis (CGD) is considered as a multifactorial pathology, in the etiology of which the leading role is assigned to infection with H. pylori (HP). The issue of symbiotic relationships of HP with other infectious agents, including pathogens of parasitic and fungal nature, is being discussed in the scientific community, but the features of combined lesions are insufficiently described. The aim of the study was to study the frequency of H. pylori and protozoal fungal invasions in chronic inflammatory diseases of the stomach and duodenum, the effect on morphological changes in the mucous membranes of the gastrointestinal tract. Material and methods. 87 patients diagnosed with CGD were examined. The material for the study was biopsies of the gastric mucosa and duodenum 12, faeces. Cytological and immunochromatographic methods were used to identify HP. The staining by Romanovsky-Giemsa was used for detection of the morphological changes in the mucous membranes and the microflora. Parasitoscopic methods were used to identify the protozoa. The results of the study. When studying the microbial composition of the stomach in 87 patients with CGD, the mixed microflora was found, including eukaryotic nature. A preliminary analysis of the results in the group of H. pylori-positive patients (n=57) showed that the association of H. pylori and yeast-like fungi of the genus Candida was found in 29% of cases, the association of H. pylori, yeast-like fungi of the genus Candida and protozoa (G. lamblia) - in 19% of cases. The study of the SOF microflora of patients with CGD showed the presence of more severe forms of morphological changes, in particular, intestinal metaplasia and dysplasia, in H. pylori and fungal-protozoal mixed invasion, than in the absence of infection (p<0.05). A significant difference was noted in the group of H. pylori -positive for the detection of protozoa and fungi of the genus Candida compared with the group of H. pylori-negative patients (p<0.05). Conclusion. The presence of protozoal fungal mixed invasion in H. pylori - associated with the gastroduodenitis can have a potentiating pathogenetic effect on the functions of all parts of the gastrointestinal tract, leading to dysbiotic disorders and the development of a chronic inflammatory process, which requires the development of algorithms for complex diagnosis and correction of dysbiotic conditions.

The article describes a clinical case of diagnosis and treatment of onychomycosis caused by combined flora — the causative agent of favus Trichophyton schoenleinii and yeast-like fungi of the Candida genus. An 85-year-old female patient was examined, who was under observation for 14 days in the 24-hour inpatient department of the State Budgetary Healthcare Institution of Bashkortostan “Republican Skin and Venereal Diseases Dispensary” in Ufa. Complaints and medical history were collected, physical examination, microscopic examination and microbiological culture of pathological material, systemic and topical treatment were performed. As a result, microscopy done twice revealed mycelium and yeast-like cells, and culture results revealed T. schoenleinii. The treatment resulted in regression of clinical manifestations. In this case, the clinical picture was characterized by a prolonged, subtle disease course, a combination of two pathogens, and a slow response to therapy. Patients’ comorbidity, elderly age, and subtle clinical symptoms are important factors of prolonged onychomycosis, multiple lesions of the nail plates, and the need for longer and combined treatment.

Background: The yeast in pindang eggs is indigenous. Isolation, characterization and identification of yeast in boiled eggs were carried out to determine the genus of yeast. The yeast has the potential to be a starter for fermented chicken eggs. This study aims to determine the characteristics of the yeast genus that has been isolated from boiled eggs and to determine the genus and species of yeast that have the best ability as a starter based on pH, total yeast count and reducing sugar content in fermented chicken eggs. Methods: This study was carried out by isolating, identifying, characterizing the yeast in pindang eggs and then carrying out chicken eggs fermentation by adding yeast with the ability to ferment sugar in chicken eggs. Result: The results show that pindang eggs are thought to contain the genera Candida, Hyphopichia and Pichia. These three yeasts have the ability to ferment sugar, making them suitable as starters for fermented chicken eggs. Pichia kudriavzevii is the best starter for fermented chicken eggs, producing the lowest pH (5.830) and the lowest reducing sugar content (0.058%) with low total yeast count (4.7 × 104 CFU/mL).