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This study aimed to determine the factors associated with the co-infection of Candida albicans and Candida glabrata. This retrospective study conducted between January 2015 and 2020 comprised 131 patients who were diagnosed with oral candidiasis. The factors associated with this condition were analyzed by univariate and multivariate logistic regression. Age and denture use were identified as significant risk factors in the univariate analysis. The multivariate logistic regression analysis revealed that denture use (odds ratio [OR], 8.44 95% confidence interval [CI], 1.99-28.3) and immunosuppressive therapy (OR, 9.20; 95% CI, 1.19 -62.0) had significant effects on co-infection with the two Candida species. These findings suggest that immunosuppressive therapy using and dentures is significantly associated with oral candidiasis caused by co-infection of C. albicans and C. glabrata. (C) 2021 Association for Dental Sciences of the Republic of China. Publishing services by Elsevier B.V.

Introduction Fungal infections have risen exponentially in the last decade. In fact, candidiasis has become the most frequent type of hospital acquired infection especially in patients receiving treatment for chronic and terminal illnesses in a hospital. A retrospective analysis for a period of twenty year was undertaken to analyze the incidence rate of candidiasis, especially of Candida species, patients treated in a tertiary care center. Materials and methods Clinical data was collected from samples of patients who were receiving tertiary care were presenting with clinically suspected fungal infections. Direct microscopy with 10% potassium hydroxide was done to visualize the presence of fungal elements, and Gram staining was done for any suspected yeast infection. The samples were inoculated on Sabouraud’s Dextrose Agar and kept at 22 °C. Results A total of 1256 samples with presumed fungal etiology were included in the study. The maximum number of fungal infections were present in elderly (70–79 years age). Females (53.8%) were more affected (45.5%). 21% isolates were identified as yeast but belonged to non-Candida fungi. Among Candida species, Candida albicans was the most dominant species (58.3%) followed by Candida glabrata (6.4%). The year-round data of fungal cases showed that the highest incident of Candida albicans infection were in January with a mean value of 3.80, while the lowest infections were reported in June, with prevalence of 2.32 of C. albicans. The twenty-year data analysis showed that the years 2001 and 2000 showed the highest incidents of C. albicans, with a mean prevalence of 7.50 and 6.83, respectively. Specimen vs fungal prevalence data showed that 38% of the C. albicans were isolated from body aspirate specimens, followed by 26% from swab specimens. Conclusion The high prevalence of Candida spp. in the present study suggests increased susceptibility of patients with critical or chronic illnesses to fungal infections.

This study aimed to evaluate the antifungal activity of grapefruit seed extract (GSE) against Candida species and to confirm the biological safety. Two-fold serial dilutions of GSE or miconazole were prepared. Cell suspensions of Candida albicans, Candida glabrata, Candida tropicalis, or Candida parapsilosis were added to each diluted solution, and the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were determined. The MFC/MIC ratio was calculated to determine whether the substance had a fungistatic or fungicidal activity. Each Candida species treated with GSE were examined using a scanning electron microscope (SEM) and fluorescence microscopy. The effect of GSE on EpiOral™, a three-dimensional tissue model, was assessed by tissue viability assay and histological analysis. GSE was applied to the mandibular gingiva of Japanese white rabbits, and the gingival tissues were histopathologically assessed. GSE demonstrated a fungicidal effect against all tested Candida species, whereas miconazole demonstrated a fungicidal effect against only C. albicans. SEM images showed various cell damage patterns in GSE-treated Candida cells. Fluorescence microscopy revealed that almost all Candida cells were killed by GSE-treated. EpiOral™ treated with GSE showed no effect on the tissue viability and there was no remarkable difference between GSE and control groups in the histological analysis. The histopathological observation in the oral mucosa treated with GSE showed no significant histopathological changes. Taken together, GSE has a fungicidal activity against C. albicans, C. glabrata, C. tropicalis, and C. parapsilosis, and no adverse effects were observed in vitro and in vivo.

Voriconazole is an antifungal agent that is commonly used to treat fungal infections. Like any drug, it can cause side effects, especially in the case of overdose. Few cases of hyperkalemia have been reported, which presented when the serum voriconazole level was elevated or when there was voriconazole-drug interaction. We present here a case of voriconazole-induced hyperkalemia, prescribed for an oral candidiasis due to Candida glabrata in a patient hospitalized for severe COVID-19 pulmonary infection. Inflammation due to COVID-19 may play a role in the variability of voriconazole concentrations and then in hyperkalemia.

Biofilms are multicellular communities where microorganisms are grown and form an extracellular matrix that protects the pathogenic microorganisms from the immunity system and antimicrobial agents. This study is aimed to identify Candida spp. isolated from urine specimens by using traditional techniques, germ tube, growth on corn meal agar medium and chromogenic medium then determine the ability of isolates to producing biofilm by tube method (TM) and congo red agar method (CRA). In our study urine specimens were obtained from 174 diabetic females in the period of six months at the Al-Wafa Specialized Center for Diabetes and Endocrinology, Mosul city, Iraq. Out of the total 174 specimens, yeast species were isolated from 56 (32.2 %) specimens. Out of the 56 isolates, 50 isolates were positive for Candida spp., especially C. glabrata which appeared maximum in 30 isolates (60 %) and followed by C. albicans 18 isolates (36%) and C. krusei 2 isolates (4 %). In the TM method for biofilm detection, C. albicans showed 16 isolates (88.9%) positive for biofilm formation followed by 29 isolates (96.7%) of C. glabrata. Furthermore, in the CRA method, all isolates (100%) of C. albicans were negative followed by 27 isolates (90%) of C. glabrata, whereas only 3 isolates (10%) of C. glabrata were positive. We can conclude that TM is the best conventional method and is sensitive to detect biofilm-forming yeast when compared with the CRA method.

A 51-year-old female with a past medical history of poorly controlled diabetes mellitus presented to the emergency department with signs of emphysematous cystitis and pyelonephritis with associated Enterococcus faecalis bacteremia and Candida Glabrata (C.Glabrata) fungemia. She was treated with micafungin and amoxicillin with improvement in her symptoms. During her course she underwent repeat imaging which showed improvement in her cystitis and no obstruction in her bladder. She was discharged on home IV micafungin. She returned to the emergency department the next day with acute urinary obstruction and was found to have a completely obstructing mucus ball in her bladder. This was removed cystoscopically and surgical cultures grew C. glabrata. She was successfully treated with intra-vesicular amphotericin B and was discharged.

Background and Purpose: Many innovations are constantly emerging with the advances in modern medicine and also it is occurring in various traditional medicines as they are considered to be safe and economical in current increasing health care financial burden. Due to emerging of MDR (multiple drug resistance) microorganisms and reduce efficacy of Modern medicines researchers and clinicians are required to revisit the traditional and alternative medicines. Material and Method: Extracts of garlic (Allium sativum), turmeric (Curcuma longa), amla (Emblica officinalis) and honey were prepared at concentration of 1mg/mL, 2mg/mL, 3mg/mL and were tested on different cancdidal species like Candida Albicans, Candida tropicalis, Candida glabrata, Candida Parapsilosis and Candida Krusei and on Fluconazole as a positive control. Disc Diffusion Test for antifungal assay was performed. Results: The anti-fungal effect was maximum at 3mg/mL concentration for all the herbal extracts. Zone of inhibition for fluconazole, was highest (35.00 ± 1.73). Among herbal extracts zone of inhibition for garlic was maximum (16.67 ± 0.88), followed by turmeric (12.67 ± 1.20), honey (11.00 ± 0.58 ), amla (9.33 ± 0.67 ). Similarly for each dose, comparing the mean zone of inhibition between antifungal groups Tukey test showed significantly (p

Background Data regarding yeast microbiota in goat milk is scarce. Aims To isolate and identify species of the genus Candida in milk samples from clinically healthy goats, and evaluate their enzymatic activity and biofilm formation. Methods 1092 milk samples from clinically healthy goats were collected and processed. The yeast isolates were identified by phenotypic, methods and their enzymatic activity (phospholipase, hemolysin and protease) and biofilm formation evaluated. Results We obtained 221 Candida isolates belonging to six species: Candida kefyr (35.7%), Candida guilliermondii (33%), Candida famata (23.5%), Candida glabrata (5.9%), Candida albicans (1.35%) and Candida parapsilosis sensu lato (0.45%). Protease activity was detected in all Candida species while hemolysin activity was only present in C. kefyr, C. guilliermondii, C. famata and C. albicans. Only C. albicans showed phospholipase activity. With the exception of C. parapsilosis sensu lato, all Candida species formed biofilm, with 60.19% of the isolates being poor producers, 9.93% moderate producers, and 1.35% strong producers. Conclusions The milk of clinically healthy goats contains several species of the genus Candida that could play a role as opportunistic pathogens in mastitis.

Candida glabrata is the second frequent etiologic agent of mucosal and invasive candidiasis. Based on the recent developments in molecular methods, C. glabrata has been introduced as a complex composed of C. glabrata, Candida nivariensis, and Candida bracarensis. The four main classes of antifungal drugs effective against C. glabrata are pyrimidine analogs (flucytosine), azoles, echinocandins, and polyenes. Although the use of antifungal drugs is related to the predictable development of drug resistance, it is not clear why C. glabrata is able to rapidly resist against multiple antifungals in clinics. The enhanced incidence and antifungal resistance of C. glabrata and the high mortality and morbidity need more investigation regarding the resistance mechanisms and virulence associated with C. glabrata; additional progress concerning the drug resistance of C. glabrata has to be further prevented. The present review highlights the mechanism of resistance to antifungal drugs in C. glabrata.

Background: Candida species are responsible for causing many health care associated and central line associated infections. They are responsible for causing opportunistic infection in human beings. Genus of Candida is composed of a heterogeneous group of organsims.Invasive infections of Candida mainly caused by Candida albicans, Candida glabrata, Candida parapsilosis & Candida tropicalis. Aim and objective: The main objective of this study was to isolates candida albicans and Non- albicans candida and their antifungal susceptibility testing. Materials and Methods: The study was carried out in the Department of Microbiology, in Tmu Hospital Moradabad. Total numbers of 806 clinical samples were processed in which 206 isolates were taken for candida. Isolation and antifungal susceptibility testing done by Vitek-2 system. Result: Out of 206 samples 77(37%) were C.albicans and 129(63%) were Non-albicans candida (NAC). Maximum isolated species were C.albicans 77(37%), followed by C.tropicalis 70(34%), C. parapsilosis 24(12%), C. glabrata 19(9%), C. dubliniensis 12(6%), C. krusei 3(1%), C.african 1(1%). Conclusion: Infection caused by NAC species have increased. C.tropicalis was the most common isolated species. Candida albicans, C.glabrata and C.krusei were shown high susceptibility to fluconazole and voriconazole. Amphotericin B, Caspofungin, Micafungin and Flucytosine shows high susceptibility towards other candida species. Keywords: Candida species, Non-albicans candida, Antifungal Susceptibility Testing

Neonates admitted to neonatal intensive care units are at a risk of developing healthcare-associated infections, leading to increased risk of mortality. This study aimed to identify organisms causing such late-onset infections in neonates and determine whether these isolates were genetically identical to those from the surrounding environmental surfaces and hands of healthcare workers (HCWs). A cross-sectional study was carried out over a period of 4 months in a university neonatal intensive care unit (NICU). Samples were collected from all neonates with symptoms of late-onset infections (n = 180). Fingerprint samples of 21 healthcare workers as well as 330 random environmental samples were also taken from the unit. Isolates from neonates, environment and fingerprints were subjected to protein electrophoresis followed by sequencing to detect genetic similarities. Almost half of neonatal samples were culture positive (91/180, 50.6%), out of which 72% of bacterial isolates (49/68) were multi-drug resistant. Klebsiella pneumoniae (32.6%) and Candida spp. (28.4%) were the commonest neonatal isolates. A cluster of two homologous Klebsiella pneumoniae strains was isolated from a neonate and an examining bed, while another homologous cluster was from a neonatal sample and a portal incubator. A third cluster was isolated from hands and three neonatal samples. This cluster (caused by Klebsiella pneumoniae strain NH54 chromosome) was found to perpetuate over the 4 months of the study. All three clusters were multi-drug-resistant Klebsiella pneumoniae. A homologous pair of each of Candida tropicalis and Candida glabrata was isolated from the blood of two neonates, and one neonatal and a crash cart sample, respectively. Overall, 8.8% (8/91) of neonatal samples were found to be homologous to other neonatal/environmental/hand isolates, denoting perpetuation of pathogens between neonates themselves and also other reservoirs of infections. Conclusion: The hands of HCWs, crash carts and incubators are reservoirs of pathogens and can lead to nosocomial infections. Clusters of multi-drug-resistant Klebsiella pneumoniae and Candida spp. were the predominant neonatal pathogens in this NICU.

The present work describes design of a small library of new 1,2,3-triazole-appended bis-pyrazoles by using a molecular hybridization approach, and the synthesized hybrids were evaluated for their antifungal activity against different fungal strains, namely, Candida albicans, Cryptococcus neoformans, Candida glabrata, Candida tropicalis, Aspergillus niger, and Aspergillus fumigatus. All the compounds exhibited broad-spectrum activity against the tested fungal strains with excellent minimum inhibitory concentration values. The molecular docking study against sterol 14α-demethylase (CYP51) could provide valuable insights into the binding modes and affinity of these compounds. Furthermore, these compounds were also evaluated for their antioxidant activity, which also resulted in promising data.

Known morpholine class antifungals (fenpropimorph, fenpropidin, and amorolfine) were synthetically modified through silicon incorporation to have 15 sila-analogues. Twelve sila-analogues exhibited potent antifungal activity against different human fungal pathogens such as Candida albicans, Candida glabrata, Candida tropicalis, Cryptococcus neoformans, and Aspergillus niger. Sila-analogue 24 (fenpropimorph analogue) was the best in our hands, which showed superior fungicidal potential than fenpropidin, fenpropimorph, and amorolfine. The mode of action of sila-analogues was similar to morpholines, i.e., inhibition of sterol reductase and sterol isomerase enzymes of ergosterol synthesis pathway.

The current study described the systematic and detailed extracellular synthesis method of silver nanoparticles (AgNPs) using Streptomyces hirsutus strain SNPGA-8 by green synthesis method. The AgNPs were subjected for characterizations using UV–Vis, FTIR, TGA, TEM, EDX, XRD, and zeta-potential analyses. The antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis, Escherichia coli, Candida albicans, Alternaria alternata, Candida glabrata and Fusarium oxysporum was determined by the agar well diffusion technique. The cytotoxicity of AgNPs against human lung cancer (A549) was studied by MTT and ROS assays and capping of proteins of AgNPs from SDS-PAGE. In the UV–Vis., absorption peak was found at 418 nm, FTIR analysis revealed the infrared bands of specific functional groups from 3273 cm−1 to 428 cm−1; TEM data confirmed the spherical shape, smallest size of particle as 18.99 nm, while EDX analysis confirmed the elemental composition of AgNPs with 22.24% Ag. The XRD pattern confirmed the nature of AgNPs as crystalline, and zeta potential peak was found at −24.6 mV indicating the higher stability. The AgNPs exhibited increased antimicrobial activity with increase in dosage volume and considerable MIC and MBC values against microbial pathogens. In the MTT cytotoxicity assay, the IC50 value of 31.41 μg/mL is obtained against A549 cell line, suggesting the potential of AgNPs to inhibit the tumour cells; and ROS assay displayed increased ROS production with increase in treatment time. Based on the results, it is evident that Streptomyces hirsutus strain SNPGA-8 AgNPs are potentially promising to be applied for biomedical uses.

Introduction: Onychomycosis, also called tinea unguium, is a common fungal infection affecting the nails. After dermatophytes, Candida species are recognized as second-line pathogens responsible for this infection. The treatment of onychomycosis requires a long time and is associated with high rates of recurrence. Antifungal medicines conjugated with gold (Au-NP) nanoparticle are the possible platforms for the reduction of drug resistance. Methods: In the present study, we reported the in-vitro antifungal activity of itraconazole (ITZ) – Au conjugates, time-kill studies, and biofilm-producing ability of six ITZ-resistant C. glabrata. Results: 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium bromide (MTT) quantitative results revealed that four out of six resistant isolates studied able to form biofilms in vitro. ITZ-Au conjugates were more effective than ITZ or Au nanoparticle alone, and the time-kill tests pointed to the suitable effect of ITZ-Au conjugate. Conclusion: The present study concluded that ITZ-Au conjugates have an inhibitory effect on the biofilm of resistant C. glabrata isolates. Further studies are needed to compare the ex-vivo onychomycosis model.

Microbial cell factories provide a sustainable and economical way to produce chemicals from renewable feedstocks. However, the accumulation of targeted chemicals can reduce the robustness of the industrial strains and affect the production performance. Here, the physiological functions of Mediator tail subunit CgMed16 at l-malate stress were investigated. Deletion of CgMed16 decreased the survival, biomass, and half-maximal inhibitory concentration (IC50 ) by 40.4%, 34.0%, and 30.6%, respectively, at 25 g/L l-malate stress. Transcriptome analysis showed that this growth defect was attributable to changes in the expression of genes involved in lipid metabolism. In addition, tolerance transcription factors CgUSV1 and CgYAP3 were found to interact with CgMed16 to regulate sterol biosynthesis and glycerophospholipid metabolism, respectively, ultimately endowing strains with excellent membrane integrity to resist l-malate stress. Furthermore, a dynamic tolerance system (DTS) was constructed based on CgUSV1, CgYAP3, and an l-malate-driven promoter Pcgr-10 to improve the robustness and productive capacity of C. glabrata. As a result, the biomass, survival, and membrane integrity of C. glabrata 012 (with DTS) increased by 22.6%, 31.3%, and 53.8%, respectively, compared with those of strain 011 (without DTS). Therefore, at shake-flask scale, strain 012 accumulated 35.5 g/L l-malate, and the titer and productivity of l-malate increased by 32.5% and 32.1%, respectively, compared with those of strain 011. This study provides a novel strategy for the rational design and construction of DTS for dynamically enhancing the robustness of industrial strains. This article is protected by copyright. All rights reserved.

Introduction. Coupled with the increasing use of indwelling vascular catheters and prosthetic cardiac valves is an uptrend in sepsis secondary to fungemia. An insidious onset often shrouds the initial diagnosis, contributing to poor outcomes. Candida infective endocarditis (CIE) is a feared complication of candidemia, associated with high mortality rates. It requires prolonged hospital stays for medical and, often, surgical management. We report a case of a massive intracardiac Candida mass in an adult with native valve CIE. Case. A 51-year-old male on chronic total parenteral nutrition (TPN) because of bowel resection presented with fevers, night sweats, and unintentional weight loss. He was febrile and tachycardiac on admission, with a benign physical examination. Laboratory workup showed elevated inflammatory markers and an acute kidney injury. Extended blood cultures showed growth of Candida glabrata (C. glabrata) and Candida dubliniensis (C. dubliniensis). Transthoracic (TTE) and transesophageal echocardiography revealed a large mobile right atrial mass (4 cm × 6 cm × 2.5 cm), extending to the right ventricular outflow tract. Since he was a poor surgical candidate, management with micafungin was initiated and continued for 8 weeks. He responded well to the regimen with resolution of the fungal mass on follow-up TTE 3 months later. In anticipation of the future need for TPN, he continues on lifelong suppressive oral fluconazole. Conclusion. CIE may be an insidious complication of indwelling central venous catheters, necessitating a high index of suspicion. Conservative management, with antifungal therapy, can yield favorable outcomes in poor surgical candidates.

Objetivo: actualizar el diagnóstico y la terapéutica en el síndrome de flujo vaginal (vaginitis / vaginosis), en mujeres gestantes y no gestantes. Materiales y métodos: se hizo una revisión sistemática y búsqueda bibliográfica de la literatura en diferentes bases de datos electrónicas, para identificar literatura relevante: Embase, Medline, Pubmed, entre otras, y libros de texto impresos; por medio de términos de búsqueda libres y estandarizados. Entre enero de 1991 y enero de 2021; sin restricción de idioma. Los comparadores incluyeron vaginosis bacteriana (VB), candidiasis vulvovaginal (CVV) e infección por Trichomonas vaginalis. Resultados: se incluyeron 93 estudios. La VB es la infección vaginal más prevalente (40-50%) en mujeres en edad reproductiva, seguida por la CVV (20-25%). La cándida albicans es la especie más prevalente (60,3%); mientras que de las especies de Cándida no albicans, la Cándida tropicalis (9,9%) y la Cándida glabrata (12,2%) son las que prevalecen. La Gardnerella vaginalis es la causante del 19,5% de VB. La tricomoniasis vaginal es la Infección de Transmisión Sexual (no Viral) más común (0,8- 3,2 %). Conclusiones: la VB es la infección vaginal más prevalente en mujeres en edad reproductiva, seguida de la CVV. La Cándida albicans es la especie más prevalente en la CVV mientras que la Gardnerella vaginalis predomina en la VB. Se requieren acciones de intervención urgentes que permitan mejorar las medidas de prevención de las infecciones vaginales, con el propósito de disminuir tanto la prevalencia como las complicaciones.

Green tea is credited as one of the world’s healthiest drinks with enriched antioxidants. It is known for its multi-beneficial health benefits against diabetes, blood pressure, hypertension, gastro-intestinal upset and is bestowed with significant antimicrobial potential. There are previous scientific evidence highlighting the antifungal potential of green tea and has identified it as a potential inhibitor of non-albicans Candida species. Lansterol 14-α demethylase (Erg 11) or CYP51 protein belongs to the cytochrome P450 monooxygenase (CYP) superfamily. Erg 11 is involved in ergosterol biosynthesis and has a significant role in azole drug resistance in Candida glabrata. The present study attempted to identify the inhibitory potential of green tea phytocompounds against inhibition of Erg 11 in Candida glabrata using bioinformatics tool viz., autodock vina software. Out of 15 green tea phytocompounds investigated, the study identified, Rutin (-10.5 kcal) Kaempferitrin (-9.4kcal), Epigallocatechin gallate (-10kcal), Epicatechin gallate (-8.7kcal), and Coumaroylquinic acid (-8.6kcal) acid as the potent phytocompounds which showed significant molecular interaction with Erg 11 in Candida glabrata. In attribution to the constant emergence of azole-resistant isolates, this preliminary analysis therefore, indicated the potential of green tea phytocompounds against inhibition of non-albicans Candida specific candidiasis. However, further, in vitro antimicrobial efficacy of these phytocompounds, the dose regime, drug likeliness, and cytotoxic analysis are required to be investigated and validated.

Background Candida auris is an emerging multidrug-resistant and highly virulent yeast that spreads easily among patients. Aims To describe the characteristics of candidemia caused by C. auris in the southeast of Spain (Autonomous Community of Valencia – ACV) through a 5-year population-based study. Methods An analysis of all the episodes of candidemia diagnosed in the ACV, with approximately 4,500,000 inhabitants, during 2013–2017, was done. Data were obtained from the Epidemiological Surveillance Valencian Network, a network that collects all the microbiological data from the hospitals in the study region. Results Based on the records, 1.9% of the isolates recovered from the positive blood cultures (corresponding to 1789 patients) were yeasts. This implies an annual rate of 7.09 cases/100,000 inhabitants. Of the 23 yeast species isolated, Candida albicans was the most frequent (37.3%), showing a higher frequency than Candida parapsilosis (28.4%) and Candida glabrata (15.6%) (p 500 beds (63.3% versus 36.7% in small hospitals) (p Conclusions The information about the local epidemiology of candidemia is essential in order to decide the best empirical treatment approach. This study reports the novel presence of C. auris in large hospitals. This pathogen has usually resistance to several antifungals and causes severe fungemia, so the results of this work reveal the need to monitor the presence of this species systematically.

The emergence of resistant fungi to available drugs highlights the need for new antifungal drugs. The present study aimed to evaluate the antifungal activity of the isolated compounds, fractions and crude extract from the leaf of Mallotus oppositifolius (Geiseler) Müll. Arg. Three pure compounds labelled 1-3 were isolated from the methylene chloride / methanol (1/1) extract of the leaf of this plant using chromatography techniques. These compounds were identified using analytical spectroscopic methods as betulinic acid (1), quercetine (2) and quercitin (3). The crude extract, fractions and compounds were tested against pathogenic yeasts (Candida albicans, Candida glabrata, Candida krusei) and dermatophytes (Trichophyton rubrum, Trichophyton soudanense, Microsporum audouinii, Microsporum langeronii) using agar well diffusion and dilution methods. The safety of the crude extract was studied on Wistar rats according to the WHO guidelines. The minimal inhibitory concentration (MIC) values ranged from 48 to 781 μg/ml against yeasts for crude extract and fractions, and 1.86 to 25000 μg/ml against dermatophytes for pure compounds, fractions and crude extract. The antifungal activity of pure compounds was not determined against yeasts. The crude extract of leaf was found to be safe in rat at up to 12 g/kg. The results achieved supported the traditional use of Mallotus oppositifolius leaf for the treatment of fungal infections.Keywords: Mallotus oppositifolius, antifungal activity, safety, phytochemical screening.

Recently, Cryptococcus and non-albicans Candida (NAC) have emerged as health-threatening pathogens for clinical fungal infections. Due to their increased resistance to existing antifungal drugs, novel antifungals are urgently needed. In this study, we evaluated the antifungal effect of VT-1161 and its comparators itraconazole and fluconazole against common fluconazole-sensitive or -resistant Cryptococcus and NAC strains. The tested strains were obtained from Chinese patients by the Invasive Fungal Infection Group within the past 2 years. The minimum inhibitory concentrations (MICs) of VT-1161 and other triazoles were measured according to the Clinical and Laboratory Standards Institute (CLSI) M27-Ed4 guidelines. We found that VT-1161 exhibited strong in vitro activity against Cryptococcus spp.. VT-1161 (geometric mean MIC = 0.024 μg/mL) was 21.7-fold and 104.5-fold more potent than itraconazole and fluconazole, respectively. Against the seven Cryptococcus neoformans isolates with higher fluconazole MICs (≥8 μg/mL based on the MIC90 value of this azole), VT-1161 maintained potent activities, with MICs ranging between 0.031 and 0.5 μg/mL. For NAC spp., VT-1161 (geometric mean MIC = 0.099 μg/mL) was 6.0-fold and 11.4-fold more effective than itraconazole and fluconazole, respectively. There is a positive correlation of the MICs between VT-1161 and itraconazole/fluconazole. The MIC values of VT-1161 against Candida glabrata and Candida tropicalis were significantly lower than those of fluconazole, whereas for Candida parapsilosis the differences in the MIC values between VT-1161 and fluconazole were not statistically significant. The results showed that tetrazole VT-1161 might be a promising candidate for treating Cryptococcus and NAC infections.

BACKGROUND: Candida is considered the most common cause of opportunistic infections in the world. Increased use of antifungal agents may have led to increasing resistance of Candida for antifungals and may be related to therapeutic failures. Recently, a multidrug-resistant Candida auris has immerged causing outbreaks in several countries all over the world. This discovered superbug is widely spread causing a broad range of health care-associated infections. AIM: This study aims to set a protocol for the identification and detection of the prevalence of C. auris in tertiary Egyptian hospitals following the center of disease and control (CDC) methodology. METHODOS: Over almost 2 years, 400 Candida isolates were collected from different wards of Cairo University Hospitals. Identification of species of all isolates was done by germ tube test followed by sub-culturing on chromogenic agar media for confirmation. Candida non-albicans isolates were further subjected to thermotolerance. Isolates that grew in 42°C were further identified by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry for definite species identification. Antifungal susceptibility using E-test was done for isolates identified by MALDI to detect resistance patterns. RESULTS: Among the 400 isolates, 227 (56.75%) were Candida albicans while 180 (43.25%) were non-albicans Candida. Candida non-albicans was classified by Chromagar as following; 25 (13.8%) were Candida tropicalis, 43 (23.8%) were Candida krusei, and 112 (62.2%) were other Candida spp. (Candida glabrata, Candida kefyr, Candida parapsilosis, and Candida lusitaniae). Using thermotolerance, 10 isolates grew at 42°C suspecting C. auris. MALDI-TOF was used for definite and final identification; five isolates were identified as C. glabrata, four as C. krusei, and one C. kefyr. Antifungal susceptibility testing of the 10 identified isolates revealed total resistance to fluconazole. CONCLUSION: Following the set protocol for identification based on CDC guidelines, C. auris is not prevalent in Egyptian hospitals. Fluconazole resistance is on the surge among candida isolates. Further studies on a bigger scale including larger number of hospitals are recommended.

We report a case of Candida glabrata endophthalmitis which was effectively treated by intravitreal liposomal amphotericin B (L-AMB) injection. A 72-year-old man was referred to our department for positive blood culture of Candida glabrata. First ophthalmologic examination revealed a chorioretinal lesion in left eye, and the patient was diagnosed as possible candida chorioretinitis. Despite systemic antifungal therapy, his chorioretinal lesion increased in both eyes and complicated by vitritis. Intravitreal administration of L-AMB was introduced for probable candida endophthalmitis. Finally, improvement of vitritis and regression of chorioretinal lesions were obtained by total of 9 times intravitreal injection. Our case suggests the safety and efficacy of intravitreal L-AMB injection for Candida glabrata endophthalmitis.

The objective of the study was to evaluate the use of targeted multiplex Nanopore MinION amplicon re-sequencing of key Candida spp. from blood culture bottles to identify azole and echinocandin resistance associated SNPs. Targeted PCR amplification of azole (ERG11 and ERG3) and echinocandin (FKS) resistance-associated loci was performed on positive blood culture media. Sequencing was performed using MinION nanopore device with R9.4.1 Flow Cells. Twenty-eight spiked blood cultures (ATCC strains and clinical isolates) and 12 prospectively collected positive blood cultures with candidaemia were included. Isolate species included Candida albicans, Candida glabrata, Candida krusei, Candida parapsilosis, Candida tropicalis and Candida auris. SNPs that were identified on ERG and FKS genes using Snippy tool and CLC Genomic Workbench were correlated with phenotypic testing by broth microdilution (YeastOne™ Sensititre). Illumina whole-genome-sequencing and Sanger-sequencing were also performed as confirmatory testing of the mutations identified from nanopore sequencing data. There was a perfect agreement of the resistance-associated mutations detected by MinION-nanopore-sequencing compared to phenotypic testing for acquired resistance (16 with azole resistance; 3 with echinocandin resistance), and perfect concordance of the nanopore sequence mutations to Illumina and Sanger data. Mutations with no known association with phenotypic drug resistance and novel mutations were also detected.

BACKGROUND: The genus Candida includes about 200 different species, but only a few are able to produce disease in humans. The species responsible for the highest proportion of human infections is Candida albicans. However, in the last two decades there has been an increase in the proportion of infections caused by other Candida species, including C. glabrata (Nakaseomyces glabrata), C. parapsilosis, C. tropicalis, C. krusei (Pichia kudriavzevi) and more recently C. auris. Decolonization of patients has been used as an infection control strategy for bacterial infections, but information about decolonization products used in clinical practice for Candida and other fungal pathogens is limited. Compounds with antimicrobial activity, such as triclosan (TR), boric acid (BA) and zinc oxide (ZO), are mainly used in personal care products. These products can be used for long periods of time without an abrasive skin effect and are a possible alternative for patient decolonization in health care settings. OBJECTIVE: The aim of this study was to evaluate the antifungal activity of boric acid (BA), triclosan (TR) and zinc oxide (ZO), individually and combined, against clinically relevant Candida species. MATERIALS AND METHODS: Compounds to be screened for antifungal activity were evaluated at different concentrations, alone, and combined, using a well diffusion assay. The statistical evaluation was performed using analysis of variance (ANOVA) and a post hoc analysis using the multiple comparisons method. RESULTS: Individually, BA and TR showed antifungal activity against all Candida species evaluated but ZO did not show any antifungal activity. Mixtures of BA [5%] - TR [0.2%]; BA [5%] - TR [0.3%]; BA [5%] - TR [0.2%] - ZO [8.6%]; and BA [5%] - TR [0.2%] - ZO [25%] yielded the highest antifungal activity. An increased antifungal effect was observed in some mixtures when compared with individual compounds. CONCLUSIONS: We demonstrated antifungal activity of BA and TR against multiple Candida species, including against a clade of the emerging healthcare-associated pathogen C. auris. Additionally, this study shows enhancement of the antifungal effect and no antagonism among the mixtures of these compounds. Further research is needed to determine whether these compounds can reduce the burden of Candida on skin.

Background Although fungi are found relatively easily by microscopic examination of pathological samples, identification of fungal genus and species in pathological samples is not easy because the morphological features of fungi are similar among genera and species. Objectives A multiple real-time PCR was developed for identification of fungal genus/species, and morphological characterizations of fungi were analyzed in pathological samples. Patients/methods Seventy-five formalin-fixed, paraffin-embedded samples morphologically proven to contain any fungus were examined. A multiple real-time PCR system was developed to identify 25 fungal genus/species in pathological samples. Morphology of fungus in the specimens was re-reviewed retrospectively based on the results of real-time PCR. Results Real-time PCR identified fungal genus/species in 56 of 75 (74.6%) specimens with histologically proven fungal infection. In 53 specimens of filamentous fungi, Aspergillus spp. (22 specimens), Cladosporium (8), Scedosporium apiospermum (4), Malassezia sympodialis (1) and Candida albicans (1) were identified. Pseudohyphae of Candida were confused with filamentous fungus in a case. Morphological observation suggested differences in the presence of septated or non-septated hyphae, the filament size, and the branch angle among genus/species of filamentous fungi; however, genus/species was not able to be determined by their morphological features. In 22 specimens of yeasts, real-time PCR allowed for the identification of Candida albicans (12 specimens), Candida glabrata (2), Cladosporium (2), Scedosporium apiospermum (2), Pichia kudriavzevii (1), and Aspergillus sydowii (1). Conclusions These data suggest that it is difficult to identify fungal genus/species by morphological features alone. Real-time PCR is useful to identify fungal genus/species in pathological samples.

Background and Aim: Different Candida species isolated in humans and animals have different types of parasite activity. The most pathogenic species is Candida albicans followed by Candida tropicalis. However, the effects of the morphology of Candida krusei, Candida guilliermondii, and Candida glabrata biofilms on the pathogenicity of these species have not been fully characterized. To the best of our knowledge, there is no literature on the effect of farnesol on rare Candida species. This study aimed to check the effect of different farnesol concentrations on the species C. krusei, C. guilliermondii, and C. glabrata compared with the strain C. albicans ATCC 10231, which has been widely studied, and is a strong producer of biofilms. Materials and Methods: We studied the morphological and densitometric parameters of biofilms produced by Candida species under the influence of the drug farnesol (Sigma-Aldrich, St. Louis, MO). We used a heart brain broth with the addition of 2% bovine blood serum in 96-well plates. To each well, we added 100 μL of C. albicans, C. krusei, C. guilliermondii, or C. glabrata culture, and 0.2-400 μM farnesol. The microliter plates were cultured with the lid closed at 37°C for 48 h. Then, the liquid was removed, and the wells were washed 3 times with 200 μL phosphate buffer solution (pH 7.3). Biofilm fixation was performed using 150 μL of 96% ethanol for 15 min. Then, the microliter plates were dried for 20 min at 37°C, a 0.5% solution of crystalline violet was added, and the plates were placed in an incubator at 37°C. After 5 min, the contents of the wells were removed, washed 3 times with 200 μL of phosphate buffer solution (pH 7.2), and dried. The dye was extracted by washing with 200 μL of 96% ethanol for 30 min. The results were obtained using a photometric analyzer of enzyme immunoassay reactions at an optical density (OD) wavelength of 450 nm. Results: All of Candida spp. strains tested were susceptible to farnesol at concentrations ranging from 0.8 to 400 μM for C. albicans, C. krusei, and C. guilliermondii, and 12.5 to 400 μM for C. glabrata. Conclusion: This study provides new insights into the use of farnesol against biofilms produced by Candida species, but further studies in vivo are necessary to evaluate the effectiveness of the reduction of OD. To the best of our knowledge, the antimicrobial activity of farnesol against C. krusei, C. guilliermondii, and C. glabrata has not been reported previously, although studies have confirmed the inhibitory effect of farnesol on the growth of different microorganisms.

Introduction Infections with Candida glabrata have recently gained worldwide attention owing to its association with long hospitalizations and high mortality rates. This problem is highlighted when the infection is associated with echinocandin resistance, which is used for first-line therapy. Echinocandin resistance is exclusively attributed to functional mutations in FKS genes, and especially in hot spot (HS) regions. Unfortunately, few studies have focused on the rapid identification of FKS mutations associated with echinocandin resistance in C. glabrata. This study was intended to evaluate and validate the use of Surveyor nuclease assay (SN) for detection of FKS gene mutations. Methods SN was evaluated against three segments of FKS1 and FKS2 genes including whole gene, regions including all HSs, and the region including only HS1. Results Our results showed that SN results are basically dependent on the type of gene as well as the segment type. Interestingly, SN can detect mutations in the region containing HS1 in both FKS1 and FKS2 genes. Furthermore, SN can detect mutations in the segment containing all HS regions for FKS1 but not FKS2. SN was unable to detect mutations in the whole FKS1 and FKS2 genes. Conclusions As far as we know, this is the first study to validate SN for rapid identification of FKS gene mutations. This assay could be used as a sample for rapid identification of mutations associated with HS1 region in FKS genes, which have a predominant role for echinocandin resistance induction in C. glabrata.

After Candida albicans, Candida glabrata is one of the most common fungal species associated with candidemia in nosocomial infections. Rapid acquisition of nutrients from the host is important for the survival of pathogens which possess the metabolic flexibility to assimilate different carbon and nitrogen compounds. In Saccharomyces cerevisiae, nitrogen assimilation is controlled through a mechanism known as Nitrogen Catabolite Repression (NCR). NCR is coordinated by the action of four GATA factors; two positive regulators, Gat1 and Gln3, and two negative regulators, Gzf3 and Dal80. A mechanism in C. glabrata similar to NCR in S. cerevisiae has not been broadly studied. We previously showed that in C. glabrata, Gln3, and not Gat1, has a major role in nitrogen assimilation as opposed to what has been observed in S. cerevisiae in which both factors regulate NCR-sensitive genes. Here, we expand the knowledge about the role of Gln3 from C. glabrata through the transcriptional analysis of BG14 and gln3Δ strains. Approximately, 53.5% of the detected genes were differentially expressed (DEG). From these DEG, amino acid metabolism and ABC transporters were two of the most enriched KEGG categories in our analysis (Up-DEG and Down-DEG, respectively). Furthermore, a positive role of Gln3 in AAA assimilation was described, as was its role in the transcriptional regulation of ARO8. Finally, an unexpected negative role of Gln3 in the gene regulation of ABC transporters CDR1 and CDR2 and its associated transcriptional regulator PDR1 was found. This observation was confirmed by a decreased susceptibility of the gln3Δ strain to fluconazole.

The objective of this article was to evaluate the effectiveness of a PDT protocol to treat fungal infections in critically ill patients. We developed a protocol using PDT as a therapy to reduce oral candidiasis in mechanically ventilated patients hospitalized in an intensive care unit (ICU). Only adults 18 + years of age, who presented clinical signs of oral candidiasis, and were mechanically ventilated were recruited to participate in the study. The protocol consisted of clinical and laboratorial procedures. Clinical procedures involved the application of the photosensitizer methylene blue 0.01% activated by a diode laser (660 nm, 90 J/cm2) for 1.5 min. Saliva samples were collected before and after the PDT session. Laboratorial procedures included counting the fungi colony-forming units (CFU) and identifying the fungi species. Ten patients met the inclusion criteria. The mean age was 62 years, and 57% were men. PDT was effective in reducing colony-forming units regardless of the Candida species. We observed an average reduction of 1.99 log10 (98.54%; SD = 1.45) in fungi CFU counting. We identified Candida tropicalis and Candida glabrata as the most common fungi species. Technological and non-invasive therapies such as PDT may benefit mechanically ventilated ICU patients with oral candidiasis. Considering the compromised health of these patients and the disadvantages conventional pharmacological treatment may have for them (e.g., systemic administration of drugs and fungal resistance), PDT rises as an alternative to treat oral candidiasis in critically ill patients. Using our protocol, we will conduct a large clinical trial as the next step.

Surface-enhanced Raman spectroscopy (SERS) is a research method in which a lack of cost-effective, versatile platforms with high enhancement factor (EF) is still a major obstacle to its widespread use. The platforms should be also easy to manufacture, stable in time (for weeks or even for months) and manufactured with a highly reproducible method. We demonstrate SERS platforms based on silicon modified on the surface by laser ablation and covered with SERS-active metal. The substrates were fabricated by a femtosecond laser, thus the method is simple, very fast and creates highly uniform SERS platforms in a large number. The platform was tested with para-mercaptobenzoic acid (p-MBA) in terms of sensitivity and reproducibility. The calculated EF was at the level of 108 and the standard deviation (SD) gives 7% for 10−6 M solution of p-MBA based on the intensity of the band at 1073 cm−1. Optimized SERS substrate also exhibits excellent stability for up to six months. We also give the proof-of-concept of using our platform and, for the first time, the SERS analysis of the most important human opportunistic fungal pathogen Candida spp. (Candida glabrata, Candida albicans SN148 and C. albicans BWP17). Finally, the chemometric analysis in the form of Principal Component Analysis (PCA) allowed to strain differentiation of Candida spp., and to distinguish the studied Candida species from Gram-positive bacterial samples with Staphylococcus aureus. Our results demonstrate that the proposed SERS platform is a perfect substrate for detection, identification and differentiation between fungal and bacterial pathogens using SERS technique.

Aim: Melanin has been linked to pathogenesis in several fungi. They often produce melanin-like pigments in the presence of L-dihydroxyphenylalanine (L-DOPA), but this is poorly studied in Candida glabrata. Methods & materials: C. glabrata was grown in minimal medium with or without L-DOPA supplementation and submitted to a chemical treatment with denaturant and hot acid. Results: C. glabrata turned black when grown in the presence of L-DOPA, whereas cells grown without L-DOPA supplementation remained white. Biophysical properties demonstrated that the pigment was melanin. Melanized C. glabrata cells were effectively protected from azoles and amphotericin B, incubation at 42°C and macrophage killing. Conclusion: In the presence of L-DOPA, C. glabrata produces melanin, increases antifungal resistance and enhances host survival.

Objective: The principal objective of the study was to synthesize and evaluate the biological activities of a novel class of 5-benzylidene substituted rhodanine derivatives as antimicrobial agents. Methods: All the synthesized compounds (D1-D10) were screened for their antimicrobial activities using microdilution methods as per the reported procedure. All compounds were evaluated as potential antimicrobial agents against gram-positive bacteria: Bacillus cereus, Staphylococcus aureus, gram negative bacteria: Escherichia coli Pseudomonas aeruginosa and Klebsiella pneumoniae Fungal cultures used in the study were Aspergillus niger, Candida albicans, Candida parapsilosis, Candida tropicalis and Candida glabrata. Results: Compound D6 showed good antifungal activity in the MIC range 16μg/ml against Candida tropicalis and Compound D10 showed good antifungal activity in the MIC range 16μg/ml against Candida glabrata. Compounds D2 and D5 showed good antibacterial activity at 32μg/ml. all the other compounds showed moderate antibacterial activity. Conclusion: Based on the above results, it can be concluded that the compounds may lead to the development of more potent antimicrobial drug candidates in the near future.

BACKGROUND We aimed to describe the epidemiology of candidemia among children in South Africa. METHODS We conducted laboratory-based surveillance among neonates (≤28 days), infants (29 days to

Background Candida bloodstream infections (BSI) became an important invasive disease in the late 20th century, in particular among immunocompromised patients. Although considerable progress has been made in the management of patients with invasive mycoses, Candida BSI are still widespread among hospitalised patients and are associated with relatively high mortality. Objectives We conducted a retrospective study to evaluate patient characteristics, incidence, species distribution and antifungal susceptibility of BSI isolates of Candida spp. as well as outcomes of Candida BSI from 2001 to 2012, before the widespread use of echinocandins. This is the first epidemiological study of Candida BSI in Slovenia so far. Methods All documented candidaemia cases from 2001 to 2012 in two major hospitals-University Medical Centre and Institute of Oncology in Ljubljana, Slovenia-were taken into consideration. Candida BSI were identified in 422 patients (250 male, 172 female). Laboratory and clinical data of these patients were retrospectively analysed. Mann-Whitney U test was used to compare continuous variables and Fisher's exact test or chi-squared test for categorical variables. Results and conclusions The average incidence of Candida BSI was 0.524/10.000 patient-days (0,317/1000 admissions); 16/422 were younger than 1 year and 251/422 patients were over 60 years old. The most commonly isolated species were Candida albicans and Candida glabrata, followed by Candida parapsilosis. Majority of the patients had a single episode of Candida BSI, multiple episodes of Candida BSI occurred in 18/434 patients (4.1%); in 25/434 patients (5.8%) mixed Candida BSI were observed. Crude 30-day case-fatality rate was 55.4%.

Candida sp. é um gênero de levedura que faz parte da microbiota do corpo humano e de animais. É considerado o principal grupo de fungos patógenos oportunistas. O uso de plantas medicinais é empregado cada vez mais nas diversas áreas. A planta Cymbopogon citratus (DC). Stapf é popularmente conhecida no Brasil por Capim-cheiroso, Capim-cidreira, Capim-santo, Capim-limão, Capim-cidró, Capim-cidrão. Este estudo teve como objetivo avaliar a atividade antifúngica do óleo essencial de Cymbopogon citratus frente a leveduras do gênero Candida sp.. O óleo essencial foi obtido por hidrodestilação das folhas pelo método do arraste a vapor. A atividade antifúngica foi verificada pelas técnicas de disco-difusão, concentração inibitória mínima e concentração letal mínima. Foram utilizadas as cepas padrão ATCC de Candida albicans, Candida dubliniensis, Candida tropicalis, Candida glabrata, Candida Krusei, Candida parapsilosis e Candida utilis. O óleo essencial apresentou atividade antifúngica frente a todas as espécies. A faixa de inibição varia de 0,281 µg/mL a 1,125 µg/mL e concentração letal varia de 0,562 µg/mL a 1,125 µg/mL. Esses achados contribuem para um futuro emprego da planta C. citratus como fitoterápico em tratamento preventivo ou alternativo contra infecções fúngicas.

In the present investigation, we have evaluated the antibiofilm potential of Bacillus licheniformis SV1 derived glycolipid against C. glabrata biofilm. Impact of isolated glycolipid on the viability of C. glabrata and on inhibiting as well as eradicating ability of its biofilm were studied. Further, morphological alterations, reactive oxygen species generation (ROS) production and transcriptional expression of selected genes (RT-PCR) of C. glabrata in response with isolated glycolipid were studied. The isolated glycolipid (1.0 mg ml−1) inhibited and eradicated C. glabrata biofilm approximately 80% and 60%, respectively. FE-SEM images revealed glycolipid exposure results in architectural alteration and eradication of C. glabrata biofilm and ROS generation. Transcriptional studies of selected genes showed that the expression of AUS1, FKS1 and KRE1 were down-regulated, while that of ergosterol biosynthesis pathway and multidrug transporter increased, in the presence of glycolipid.

Background: Intensive studies and experiments have proven that the drug resistance of microorganisms is evolving very disturbingly. As a consequence, new avenues of research become strongly essential to expand knowledge on this phenomenon and develop highly effective alternatives targeting it. Therefore, there is increasing interest in the use of medicinal plants for treatment of many infections. Objective: In accordance with these findings, this study aims to evaluate the in vitro activities of fruit oil extracted from Tunisian varieties of Pistacia lentiscus, growing in Rimel forest (Bizerte) in north of Tunisia, against some medically important and emerging species of bacteria and yeasts. Methods: The antimicrobial activities were performed using the disc diffusion method. We evaluate antimicrobial activities of this oil against many species like Staphylococcus aureus; Pseudomonas aeruginosa; Escherichia coli and C. albicans and to our knowledge, in our current study, we demonstrated for the first time his activities against the following species: Listeria innocua; Salmonella enterica; Enterococcus faecalis; Shigella flexneri; Candida parapsilosis; Candida tropicalis and Candida glabrata. Results: The results showed positive activities of fruit oil against all tested bacteria and yeasts with some differences depending on the microorganism tested. The maximum antibacterial activities were obtained against Staphylococcus aureus and Pseudomonas aeruginosa. For yeasts, the oil exhibited high activity against C. parapsilosis. Conclusion: This study elaborately confirmed that Tunisian P. lentiscus fruit oil contains compounds that can be used to treat many infections.

Aims The aim of this study was to evaluate the formation of biofilm by Candida spp. isolated from the bloodstream, using traditional spectrophotometric methodologies. In addition, the goal was to compare the results with those obtained through MALDI-TOF/MS, as well as to verify its use as a potential tool for the detection of biofilm-forming strains. Methods and results Hundred and thirteen isolates of Candida spp. were studied: 41 were Candida albicans, 27 C. tropicalis, 18 C. glabrata, 17 C. parapsilosis and 10 C. krusei. Metabolic activity was determined through the tetrazolium salt (XTT) reduction assay and biomass by staining with Crystal Violet. All isolates were able to form biofilm, 94% of which were strong producers, with high biomass quantification (95%; 107/113) and high metabolic activity (99%; 112/113). Mass spectra of the biofilm-producing isolates showed differences in the intensity of mass peaks when compared with the spectra of the nonproducing strains. Conclusions It was demonstrated that MALDI-TOF/MS was able to detect specific biofilm proteins, as the mass spectra of the isolates presented differences when compared with nonproducing strains. Significance and impact of the study MALDI-TOF/MS can become a valuable tool for biofilm detection at the moment of the identification of the microorganism, thus contributing greatly to the management of patients with Candidemia.

This research study tested the protective effect of prolonged exposure of shrimp food supplemented with glucans from shrimp-pond related yeasts on shrimp Penaeus vannamei reared under the presence of the white spot syndrome virus (WSSV). The glucans extracted and purified from isolated marine yeasts identified as Debaryomyces hansenii , Candida tropicalis , Candida humilis , Candida glabrata , Pichia kudriavzevyi , Wickerhamomyces anomalus and the terrestrial Saccharomyces cerevisiae yeast were characterized by the Fourier-transform infrared spectroscopy and proton nuclear magnetic resonance spectroscopy. The treatments were prepared with food enriched with the yeast beta-glucans and the control groups without beta-glucans. Shrimp were fed thrice a day and challenged orally with WSSV on days 31, 54, 66 and muscularly at day 70. The animals were assessed for the protective effect in terms of post-infection total hemocyte counts, and survival rate. The results indicated that marine yeasts possessed s-1,3/1,6-glucans, and that D. hansenii was an excellent source yielding 30% of its dry biomass of pure glucans. For the positive control group where no glucans were added, WSSV challenges showed 100% survival when the virus was provided orally, and 40% when the virus was injected. These results also indicated that the shrimp line selected for this study was a resistant line for WSSV. Shrimp groups fed with glucans of the marine yeasts D. hansenii and C. humillis showed a significant protection, allowing shrimp survival of 66% while terrestrial yeast showed 57.14%. These results indicated that marine yeasts growing in the shrimp pond were an excellent source of beta-glucans that allowed extra protection against the mortality caused by this pathogenic virus.

Background Sodium-glucose co-transporter 2 (SGLT2) inhibitors are novel hypoglycemic agents which reduce reabsorption of glucose at the renal proximal tubule, resulting in significant glycosuria and increased risk of genital mycotic infections (GMI). These infections are typically not severe as reported in large systematic reviews and meta-analyses of the medications. These reviews have also demonstrated significant cardiovascular benefits through other mechanisms of action, making them attractive options for the management of Type 2 diabetes mellitus (T2DM). We present two cases with underlying abnormalities of the urogenital tract in which the GMI were complicated and necessitated cessation of the SGLT2 inhibitor. Case presentations Both cases are patients with T2DM on empagliflozin, an SGLT2 inhibitor. The first case is a 64 year old man with Candida albicans balanitis and candidemia who was found to have an obstructing renal calculus and prostatic abscess requiring operative management. The second case describes a 72 year old man with Candida glabrata candidemia who was found to have prostatomegaly, balanitis xerotica obliterans with significant urethral stricture and bladder diverticulae. His treatment was more complex due to fluconazole resistance and concerns about urinary tract penetration of other antifungals. Both patients recovered following prolonged courses of antifungal therapy and in both cases the SGLT2 inhibitor was ceased. Conclusions Despite their cardiovascular benefits, SGLT2 inhibitors can be associated with complicated fungal infections including candidemia and patients with anatomical abnormalities of the urogenital tract may be more susceptible to these infections as demonstrated in these cases. Clinicians should be aware of their mechanism of action and associated risk of infection and prior to prescription, assessment of urogenital anatomical abnormalities should be performed to identify patients who may be at risk of complicated infection.

Global antimicrobial crisis and advent of drug resistant fungal strains has substantially distressed disease management for clinicians. Biodegradable silver nanoparticles (AgNps) emerge as an excellent alternative remedial option. In the current study, the anti-biofilm activity of microwave irradiated kappa-carrageenan (CRG) capped AgNps against Candida albicans, and Candida glabrata was investigated in terms of their effect on reactive oxygen species (ROS) generation, cellular morphology, biochemical composition, and the activity of enzymes of extracellular matrix. Minimum inhibitory concentration and fungicidal concentration value of CRG-AgNps against both Candida spp. ranged between 400 and 500 μg/mL. The 80% of Candida biofilm was inhibited and eradicated by CRG-AgNps at a concentration of ~300 μg/mL. Microscopic studies indicate that CRG-AgNps caused morphological damage through membrane disruption and pore formation. Further, CRG-AgNps generated ROS in a concentration-dependent manner and modulated the composition of Candida biofilm ECM by increasing the carbohydrate and eDNA content. CRG-AgNps also significantly inactivated the hydrolytic enzymes, thus hindering the biofilm forming ability. In conclusion, all these results suggest that the CRG-AgNps are potential antifungal agents against Candida biofilms, and they inhibit/eradicate the fungal biofilms through multiple signalling mechanisms.

How cells respond to DNA damage is key to maintaining genome integrity or facilitating genetic change. In fungi, DNA damage responses have been extensively characterized in the model budding yeast Saccharomyces cerevisiae, which is generally not pathogenic. However, it is not clear how closely these responses resemble those in fungal pathogens, in which genetic change plays an important role in the evolutionary arms race between pathogen and host and the evolution of antifungal drug resistance. A close relative of S. cerevisiae, Candida glabrata, is an opportunistic pathogen that displays high variability in chromosome structure among clinical isolates and rapidly evolves antifungal drug resistance. The mechanisms facilitating such genomic flexibility and evolvability in this organism are unknown. Recently we characterized the DNA damage response of C. glabrata and identified several features that distinguish it from the well characterized DNA damage response of S. cerevisiae. First, we discovered that, in contrast to the established paradigm, C. glabrata effector kinase Rad53 is not hyperphosphorylated upon DNA damage. We also uncovered evidence of an attenuated DNA damage checkpoint response, wherein in the presence of DNA damage C. glabrata cells did not accumulate in S-phase and proceeded with cell division, leading to aberrant mitoses and cell death. Finally, we identified evidence of transcriptional rewiring of the DNA damage response of C. glabrata relative to S. cerevisiae, including an upregulation of genes involved in mating and meiosis – processes that have not been reported in C. glabrata. Together, these results open new possibilities and raise tantalizing questions of how this major fungal pathogen facilitates genetic change.

Made available in DSpace on 2022-04-29T08:30:53Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-07-01 Previous studies showed that the crude extract obtained from Streptococcus mutans inhibited the growth of Candida albicans reference strains. In this study, we evaluated whether the antifungal effects of S. mutans extract can be extended to clinical Candida isolates, including C. albicans and non-abicans strains with different susceptibilities to fluconazole. We verified that S. mutans extract increased the survival of Galleria mellonella larvae infected with C. albicans and C. glabrata and inhibited the fungal cells in hemolymph. These antifungal effects occurred for both fluconazole-susceptible and fluconazole-resistant strains. However, larvae infected by C. krusei were not affected by S. mutans extract. Lay Summary: Streptococcus mutans crude extract shows antifungal effects on clinical Candida strains susceptible and resistant to fluconazole in Galleria mellonella model. Department of Biosciences and Oral Diagnosis Institute of Science and Technology Sao Paulo State University (UNESP) Sao Jose Dos Campos Department of Organic Chemistry Institute of Chemistry Sao Paulo State University (UNESP) Department of Biosciences and Oral Diagnosis Institute of Science and Technology Sao Paulo State University (UNESP) Sao Jose Dos Campos Department of Organic Chemistry Institute of Chemistry Sao Paulo State University (UNESP)

The main aims of this investigation were the isolation of dimeric naphthoquinones, a new class of dinaphthodiospyrols (1-7), from chloroform fractions and screening them for antibacterial, antifungal, and antioxidant potential. The susceptibility of the isolated compounds, namely, dinaphthodiospyrol A (1), dinaphthodiospyrol B (2), dinaphthodiospyrol C (3), dinaphthodiospyrol D (4), dinaphthodiospyrol E (5), dinaphthodiospyrol F (6), and dinaphthodiospyrol G (7) was assessed for antibacterial potential using well diffusion methods. The isolated compounds showed excellent antibacterial activity against selected bacterial strains, including Gram-positive Bacillus subtilis, Streptococcus epidermis, and Bacillus subtilis, and Gram-negative bacteria Klebsiella pneumonia with the zones of inhibition 6 to 26 nm. The standard drug Imipenem showed a maximum inhibitory zone 30 to 35 nm. Similarly, the isolated compounds were screened for antifungal properties, which showed an excellent reduction in the growth of selected fungal strain including Candida albicans, Aspergillus flavus, Fusarium solani, Trichyton logifusus, Microsporum canis , and Candida glabrata. Among all the screened compounds, 7 exhibited good activity (30-49 mm), followed by compounds 5 and 6, (35-46 mm), while compounds 1-4 showed a moderate effect (8-28 mm) against the selected fungal strain against miconazole which showed potent effects (101-110.98 mm). The isolated compounds were also screened for 1, 1-diphenyl-2-picrylhydrazyl (DPPH) activity. In vitro-based free radical was employed using ascorbic acid as a standard antioxidant. The tested compounds (1-7) exhibited significant antioxidant activity in a concentration-dependent manner. The dinaphthodiospyrol 7 exhibited 97.32% scavenging activity, followed by dinaphthodiospyrol 6, 92.01%, and compounds 5 and 4 with 89.90 and 88.43% scavenging activity at 100 μg/mL, respectively; ascorbic acid showed 96.45% scavenging effect. Furthermore, docking analysis was performed to know the exact binding mode of the tetra-substituted derivatives of dinaphthodiospyrols to the selected target proteins. From the docking analysis, it was found that the docking results are well correlated with the experimental observations. In conclusion, the dinaphthodiospyrols exhibited excellent antibacterial, antifungal, and free radical scavenging potential.

Candida glabrata is an opportunistic human fungal pathogen and is frequently present in the human microbiome. It has a high relative resistance to environmental stresses and several antifungal drugs. An important component involved in microbial stress tolerance is trehalose. In this work, we characterized the three C. glabrata trehalase enzymes Ath1, Nth1 and Nth2. Single, double and triple deletion strains were constructed and characterized both in vitro and in vivo to determine the role of these enzymes in virulence. Ath1 was found to be located in the periplasm and was essential for growth on trehalose as sole carbon source, while Nth1 on the other hand was important for oxidative stress resistance, an observation which was consistent by the lower survival rate of the NTH1 deletion strain in human macrophages. No significant phenotype was observed for Nth2. The triple deletion strain was unable to establish a stable colonization of the gastrointestinal (GI) tract in mice indicating the importance of having trehalase activity for colonization in the gut. ispartof: VIRULENCE vol:12 issue:1 pages:329-345 ispartof: location:United States status: published

Currently, opportunistic fungi of the genus Candida are the main causative agents of mycoses, which are especially severe upon condition of acquired immunodeficiency. The main target for the development of new antimycotics is the cytochrome P450 51 (CYP51) of the pathogenic fungus. Due to the widespread distribution of Candida strains resistancy to inhibitors of the azole class, the screening for CYP51 inhibitors both among non-azole compounds and among clinically used drugs repurposing as antimycotics is becoming urgent. To identify potential inhibitors from the non-azole group, an integrated approach was applied, including bioinformatics analysis, computer molecular modeling, and a surface plasmon resonance (SPR) technology. Using in silico modeling, the binding sites for acetylsalicylic acid, ibuprofen, chlorpromazine and haloperidol (this compounds, according to the literature, showed antimycotic activity) were predicted in the active site of CYP51 of Candida albicans and Candida glabrata. The Kd values of molecular complexes of acetylsalicylic acid, ibuprofen and haloperidol with CYP51, determined by SPR analysis, ranged from 18 μM to 126 μM. It was also shown that structural derivatives of haloperidol, containing various substituents, could be positioned in the active site of CYP51 of Candida albicans with the possible formation of coordination bonds between the hydroxyl groups of the derivatives and the iron atom in the heme of CYP51. Thus, the potential basic structures of non-azole compounds have been proposed, which can be used for the design of new CYP51 inhibitors of Candida fungi.V nastoiashchee vremia uslovno-patogennye griby roda Candida iavliaiutsia osnovnymi vozbuditeliami mikozov, kotorye osobenno tiazhelo protekaiut pri nalichii priobretennogo immunodefitsita. Glavnoĭ mishen'iu dlia razrabotki novykh antimikotikov iavliaetsia tsitokhrom R450 51 (CYP51) patogennogo griba. Vsledstvie shirokogo rasprostraneniia rezistentnykh shtammov Candida k ingibitoram iz klassa azolov aktual'nost' priobretaet poisk ingibitorov CYP51 kak sredi soedineniĭ neazol'noĭ prirody, tak i sredi primeniaemykh v klinike lekarstvennykh sredstv, repozitsioniruemykh v kachestve antimikotikov. Dlia identifikatsii potentsial'nykh ingibitorov iz gruppy soedineniĭ neazol'noĭ prirody byl primenen kompleksnyĭ podkhod, vkliuchaiushchiĭ bioinformaticheskiĭ analiz, komp'iuternoe molekuliarnoe modelirovanie i tekhnologiiu poverkhnostnogo plazmonnogo rezonansa (SPR). S pomoshch'iu in silico modelirovaniia byli predskazany saĭty sviazyvaniia atsetilsalitsilovoĭ kisloty, ibuprofena, khlorpromazina, galoperidola (kotorye po literaturnym dannym proiavliali antimikoticheskuiu aktivnost') v aktivnom tsentre CYP51 Candida albicans i CYP51 Candida glabrata. SPR analiz pozvolil ustanovit' znacheniia Kd kompleksov atsetilsalitsilovoĭ kisloty, ibuprofena i galoperidola s CYP51, kotorye byli v diapazone ot 18 mkM do 126 mkM. Takzhe bylo pokazano, chto strukturnye proizvodnye galoperidola, soderzhashchie razlichnye zamestiteli, mogut pozitsionirovat'sia v aktivnom tsentre CYP51 Candida albicans s vozmozhnym obrazovaniem koordinatsionnykh sviazeĭ mezhdu gidroksil'nymi gruppami proizvodnykh i atomom zheleza v geme CYP51. Takim obrazom, byli predlozheny potentsial'nye bazovye struktury neazol'nykh soedineniĭ, kotorye mogut byt' osnovoĭ dlia sozdaniia novykh ingibitorov CYP51 gribov roda Candida.