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In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established. Resumo No atual contexto de patógenos fúngicos resistentes emergentes tais como Candida albicans e Candida parapsilosis, a descoberta de novos agentes antifúngicos é uma questão urgente. Esta pesquisa teve como objetivo avaliar o potencial antifúngico da 2-cloro-N-fenilacetamida contra cepas clínicas de C. albicans e C. parapsilosis resistentes a fluconazol. A atividade antifúngica da substância foi avaliada in vitro através da determinação da concentração inibitória mínima (CIM), concentração fungicida mínima (CFM), ruptura e inibição da formação de biofilme, ensaios de sorbitol e ergosterol, e associação entre esta molécula e antifúngicos comuns, anfotericina B e fluconazol. O produto teste inibiu todas as cepas de C. albicans e C. parapsilosis, com uma CIM variando de 128 a 256 µg.mL-1, e uma CFM de 512-1,024 µg.mL-1. Também inibiu até 92% da formação de biofilme e causou a ruptura de até 87% de biofilme pré-formado. A 2-cloro-N-fenilacetamida não promoveu atividade antifúngica pela ligação ao ergosterol da membrana celular fúngica, tampouco danificou a parede celular. Antagonismo foi observado ao combinar esta substância com anfotericina B e fluconazol. A substância exibiu atividade antifúngica significativa ao inibir tanto as células planctônicas quanto o biofilme das cepas resistentes ao fluconazol. Sua combinação com outros antifúngicos deve ser evitada e seu mecanismo de ação deve ser estabelecido.

The goal in this case report is to discuss one presentation of the microbiology of theCandidaspecies in an intra-abdominal abscess which is under reported. In general, theCandidaspecies fungi is a common yeast found human beings as part of the microbiology. Common sites of candidiasis are often the oral cavity or the genitalia.Candidaspecies have been found to have a mutualistic relationship with different species with the body [5]. When overproduction ofCandidaspecies occurs, it can cause pathological infections in patients. While it is common to see patients withCandidaspecies in their gut micro biome, it is not well documented to have intra-abdominal abscesses that growCandidaspecies. Typically, bacterial causes are first on the differential for clinicians. This case presentation looks to discuss some of presenting factors seen in intra-abdominal candidiasis (IAC) and what leads to it being under diagnosed.

Background: Candidaemia is the commonest fungal bloodstream infection in hospitalised patients. Diabetes is one of the risk factors for mortality from candidaemia. Methods: We compared the epidemiology, clinical characteristics and management of candidaemia in patients with and without diabetes. Results: Over a 10-year period, 200 episodes of Candida bloodstream infection were documented. Patients with diabetes were younger (58.7 vs 65.5 years), less likely to be suffering from cancer (21.8% vs 36%), and had significantly lower 30-day and 90-day crude mortality (17.2% vs 35.6% and 28.4% vs 48.6%, respectively). Candida glabrata was more common in patients with diabetes (39.3% vs 29.7%). Based on European Confederation of Medical Mycology (ECMM) quality indicators, the management of patients with and without diabetes was similar. Discussion: Our study highlights the importance of epidemiological data in relation to candidaemia in patients with diabetes and the growing threat of invasive C. glabrata infection in this subset of patients.

The detection of fungi in the human respiratory tract may represent contamination, colonization or a respiratory infection. To develop effective management strategies, a more accurate and comprehensive understanding of the lung fungal microbiome is required. Therefore, the objective of the present study was to define the “mycobiome” of mechanically ventilated patients admitted to an intensive care unit (ICU) using broncho-alveolar aspirate (“sputum”) samples and correlate this with clinical parameters and the bacterial microbiota. To this end, the mycobiome of 33 sputum samples was analyzed by Internal Transcribed Spacer2 (ITS2) amplicon sequencing of the ribosomal operons. The results show that in the investigated sputa of mechanically ventilated patients Candida spp. were most frequently detected, independent of pneumonia or antimicrobial therapy. The presence of Candida excluded in most cases the presence of Malassezia, which was the second most-frequently encountered fungus. Moreover, a hierarchical clustering of the sequence data indicated a patient-specific mycobiome. Fungi detected by culturing (Candida and Aspergillus) were also detected through ITS2 sequencing, but other yeasts and fungi were only detectable by sequencing. While Candida showed no correlations with identified bacterial groups, the presence of Malassezia and Rhodotorula correlated with oral bacteria associated with periodontal disease. Likewise, Cladosporium correlated with other oral bacteria, whereas Saccharomyces correlated more specifically with dental plaque bacteria and Alternaria with the nasal-throat-resident bacteria Neisseria, Haemophilus and Moraxella. In conclusion, ITS2 sequencing of sputum samples uncovered patient-specific lung mycobiomes, which were only partially detectable by culturing, and which could be correlated to specific nasal-oral-pharyngeal niches.

Candida auris is a nosocomial fungal pathogen of prime importance due to its global emergence and rapid spread in healthcare facilities worldwide. One important concern is that routine, conventional methods fail to identify C. auris. While molecular and protein-based assays accurately detect/identify C. auris, these methods are time-consuming, expensive, and require expertise. Therefore, the objective of the present study was to assess the potential use of a novel chromogenic medium, CHROMagar™ Candida Plus, as an economical alternative to expensive and laborious diagnostic tests. We compared CHROMagar™ Candida Plus with the standard enrichment (salt Sabouraud Dulcitol broth) medium to test the recovery efficiency of C. auris from surveillance samples. We also tested CHROMagar™ Candida Plus for its ability to distinguish C. auris from other yeast species. One hundred surveillance samples were cultured on CHROMagar™ Candida Plus and Dulcitol broth and incubated at 37 °C and 40 °C, respectively. Additionally, 32 Candida and yeast species were cultured on CHROMagar™ Candida Plus at 37 °C for three days to rule out any close resemblance to C. auris. Of 100 surveillance samples tested, 69 yielded presumptive positive C. auris exhibiting creamy pink colonies with a blue halo on CHROMagar™ Candida Plus within three days of incubation, and MALDI-TOF MS confirmed all by day 4. On the other hand, 69 of 100 surveillance samples yielded turbidity in Dulcitol broth by days 3-14 with final MALDI identification by days 5 to 17. Both media failed to identify one sample each, resulting in assay sensitivity and specificity of 99% and 97%, respectively. Of Candida and yeast species tested, 75-80% of C. metapsilosis and C. orthospilosis were misidentified as C. auris. However, previous studies indicated that these species are rarely detected in surveillance screening of C. auris. Naganishia diffluens also resembled C. auris, although it required different temperature growth (30 °C). In conclusion, CHROMagar™ Candida Plus provides rapid presumptive identification of C. auris. It would be another valuable tool in surveillance efforts to control the spread of C. auris in healthcare.

The present study aimed to compare the oral Candida rate between infected and uninfected children with the human immunodeficiency virus (HIV), as well as analyze the association between Candida spp. and predisposing factors of colonization, like oral biofilm index, caries experience, and laboratory markers of AIDS progression. A cross-sectional study was employed. Candida species were identified and quantified from saliva samples of 50 HIV-infected and 50 uninfected children. Biofilm index and decayed, missing, and filled teeth (dmft/DMFT) indices were assessed by oral clinical examinations. Additionally, CD4

The last decade has witnessed the rise of an extremely threatening healthcare-associated multidrug-resistant non-albicans Candida (NAC) species, Candida auris. Since besides target alterations, efflux mechanisms contribute maximally to antifungal resistance, it is imperative to investigate their contributions in this pathogen. Of note, within the major facilitator superfamily (MFS) of efflux pumps, drug/H

Candida albicans promotes biofilm formation on dentures, which compromises the use of poly(methyl-methacrylate) (PMMA) as a dental material. Farnesol (FAR), a natural compound that prevents C. albicans filamentation and biofilm formation, was incorporated into the PMMA matrix, to obtain antifungal PMMA_FAR materials. The tested concentrations (0·0125% and 0·4%) of FAR, 24 h after incubation on YPD agar, inhibited filamentation of C. albicans. PMMA was modified with different FAR concentrations (3–12%), and physicochemical properties, antifungal activity and cytotoxicity of these modified materials (PMMA_FAR) were tested. The presence of FAR in PMMA_FAR composites was verified by Fourier-transform infrared spectroscopy (FT-IR). Incorporation of FAR into the polymeric matrix significantly decreased hydrophilicity at all tested concentrations and significantly reduced biofilm and planktonic cells metabolic activity in the early stage of biofilm formation at ≥6% FAR in PMMA. PMMA_FAR composites with

Background: Emerging pathogen Candida auris has been associated with nosocomial outbreaks demonstrating widespread antifungal resistance. This microorganism is associated with systemic infections with a high mortality rate, and studies that contribute to a better understanding of this agent are important. Objectives: The present article aimed to carry out bibliographic research on Candida auris. Therefore, a literature search was carried out between January 2018 and January 2022, applying the following: C. auris; Candida auris and Infection, in Portuguese, English and Spanish. This review considers the available literature on C. auris and highlights key unknowns, which will provide guidance for future work in this field. Results: The data collected in the present review allowed us to divide the theme into topics on Pathophysiology, Epidemiology/clinical and drug resistance mechanisms, so that the understanding of Candida auris can be better detailed and understood. Conclusion: The predilection of C. auris by the most vulnerable and immunosuppressed patients or with comorbidities and with the potential to cause invasive and bloodstream infections with a propensity to cause outbreaks and concerns regarding resistance to antifungal agents, the fungus Candida auris, is of increasing clinical relevance. The increase in the number of detected cases and the occurrence of strains with multiple drug resistance prove to be worrisome, and applied research is essential to contribute to the knowledge of this strain and effective therapy.

We aimed to detect possible changes in Candida species distribution over time and to know the antifungal susceptibility profile of isolates obtained from patients with bloodstream infection (BSI) due to this pathogen. Risk factors associated with 30-day mortality were also assessed. We conducted a retrospective cohort study of patients diagnosed with Candida BSI at a Japanese university hospital from 2013 to 2021. The change in the distribution pattern of the Candida spp. isolated was examined by considering three successive sub-periods of 3 years each. Risk factors for 30-day mortality were determined using Cox regression analysis. In the entire study period, Candida albicans was the most frequent species (46.7%), followed by Candida glabrata (21.5%) and Candida parapsilosis (18.7%). There was no change in Candida species distribution comparing the three sub-periods analyzed. All isolates were susceptible to micafungin, and most were susceptible to fluconazole, except for C. glabrata. No isolates were resistant to amphotericin B or voriconazole. The overall 30-day mortality was 40.2%. Univariate analysis revealed an association between 30-day mortality and central venous catheter (CVC) removal at any time, high Pitt bacteremia score (PBS), and high Charlson comorbidity index (CCI). Multivariate Cox analysis found that high PBS was the only independent predictor of 30-day mortality; subsequent multivariate Cox regression demonstrated that early CVC removal significantly reduced 30-day mortality. Candida species distribution and antifungal susceptibility profile in our hospital remained similar from 2013 to 2021. Early CVC removal may improve candidemia outcomes.

Aim The study aimed to profile the volatile phytocomposition of snow mountain garlic (SMG) compared to normal garlic and investigate the anti-Candida efficacy against clinically relevant multi-drug resistant isolates of Candida species. Methods and Results Herein, SMG has shown significantly superior fungicidal power at 2x-MIC dose against C. albicans and C. glabrata in killing kinetic evaluation unlike the fungistatic effect of normal garlic. GC–MS headspace-based profiling of SMG showed 5 unique volatile compounds and a 5-fold higher content of saponins than normal garlic. In an in-silico analysis, cholesta-4,6-dien-3-ol,(3-beta) was uniquely identified in SMG as a potential inhibitor with high binding affinity to the active site of exo-1,3-betaglucan synthase, an established anti-candida drug target crucial for the biofilm matrix formation, thus suggesting a plausible anti-Candida mechanism. Conclusion The in-vitro and in-silico studies have demonstrated the Candida-cidal and anti-biofilm activities of SMG, distinguishing it from the Candida-static efficacy of normal garlic. Significance and Impact of the study This is the first report that identifies several phytochemical signatures of SMG along with a potential anti-Candida compound, that is cholesta-4,6-dien-3-ol,(3-beta)-, which appears worthy of detailed studies in the future to explore the utility of SMG as a fungal phytotherapy agent, especially against drug-resistant Candida sp.

Background: Candida auris has been associated with rapid transmission and high mortality. A novel PCR based surveillance programme was initiated at a London teaching hospital from January 2018. The results of this implementation until March 2019 are presented along with the clinical, transmission and phylogenetic characteristics encountered in that setting. Methods: A real time-PCR assay for C auris was developed, validated, and implemented for direct use on skin swabs and urine. Environmental swabs were also tested by PCR as an emergency outbreak control measure. Clinical risk factors and outcomes of patients were determined. Environmental dispersal was assessed using 24 h settle plate cultures around 9 colonised patients followed by air sampling around one colonised patient during high and low turbulence activities. Sequencing was performed using Illumina HiSeq and maximum likelihood phylogenies were constructed using rapid bootstrap analysis. Results 21 C. auris colonised patients were identified. Median turnaround time of colonisation detection reduced from 141 h (5.8 d) to approximately 24 h enabling rapid infection control precautions. Settle plates detected 70 to 600 CFU/m2 around colonised patients over 24 h and air sampling suggested dispersal during turbulent activities. C. auris DNA was detected from 35.7% environmental swabs. Despite being in a high-risk setting, no patients developed invasive infection. Sequencing analysis of isolates from this centre identified two introductions of the South Asian (Clade I) and one of the South African (Clade III) strain. Conclusion: The PCR offers a rapid, scalable method of screening and supports clinical risk reduction in settings likely to encounter multiple introductions.

Invasive candidiasis is a severe infectious disease that could lead to mortality in critically ill children.We collected data regarding demographics, underlying diseases, predisposing factors, outcomes for pediatric patients with candidemia at a medical centre in Taiwan from 2011 to 2017.Fifty-eight patients with 60 candidemia episodes were diagnosed. The 3 most common species were Candida albicans (42%), Candida parapsilosis (25%) and Candida tropicalis (23%). C. parapsilosis predominantly infected infants and neonates (median age: 0.8 years, range: 0.1-14.5). Cases with C. tropicalis had significantly higher rates of multidrug resistance (p = 0.011) and disseminated candidiasis (p = 0.025) compared with other cases. The all-cause mortality rate was 43%, and the candidemia-related mortality rate was 29%. Pediatric sequential organ failure assessment score8 [adjusted odds ratio (aOR) 66.2, 95% CI 4.03-1088.5] and posaconazole resistance (aOR 33.57, 95% CI 1.61-700.3) were the most significant risk factors associated with candidemia-related mortality, whereas treatment with effective antifungal agents within 48 h (aOR 0.07, 95% CI 0.01-0.9) was the only significant protective factor.Candidemia-related mortality was related to azole resistance; therefore, empirical therapy with echinocandin or amphotericin B is recommended pending species and susceptibility results.

Introduction: We sought to evaluate secondary infections (SIs) in patients admitted to the intensive care unit (ICU) for COVID-19 with respect to incidence, causative pathogens, and clinical outcomes. Methodology: In this two-centre retrospective study, we analysed 146 patients (96 males, 50 females; median age, 64 years) admitted to the ICU with COVID-19 between March 26 and December 31, 2020. Inclusion criteria were an ICU admission for at least 48 hours and age beyond 18 years. Patients with and without SIs were compared and the impacts of SIs and carbapenem resistance on mortality were analysed. Results: During ICU admission, 84 episodes of SIs developed in 58 patients (39.7%). A total of 104 isolates were recovered, with Gram-negative bacteria most frequent accounting for 74%. At least one carbapenem-resistant pathogen (n = 61) was recovered in 41 patients (70.1%). In multivariate analysis, the use of ECMO and an elevated procalcitonin level were significantly associated with the development of SIs. The mortality rate and the incidence of carbapenem resistance did not differ significantly in COVID-19 patients with and without SIs (p = 0.059 and p = 0.083, respectively). Conclusions: The incidences of SIs and carbapenem resistance among COVID-19 patients were alarming, emphasizing stricter infection control measures in the ICU setting. © 2022 Journal of Infection in Developing Countries. All rights reserved.

Objectives: Candida species are the most common fungal infectious agents. Candida species are important with their increasing frequency in hospital-acquired infectious agents. The issue of antibiotic resistance, which frequently encountered in bacterial agents, is unfortunately also valid in fungal infections. In the present study, we aimed to determine the resistance of Candida species in our hospital including 1350 patient beds, the materials and units of production, and their sensitivity to antifungal drugs, in particular fluconazole. Methods: Yeast growths, colony morphology, germ tube formation and VITEK 2 Compact automated identification system detected in the samples evaluated in the central laboratory of our hospital between January 1, 2021 and December 31, 2021 were typed. Antifungal susceptibilities, especially fluconazole, caspofungin, and amphotericin B susceptibilities, were determined by an automated system. Results: In total, 2446 within the Candida growing sample was determined as 49% Candida albicans, 26.9% Candida parapsilosis, %17.9 Candida tropicalis, Candida glabrata and Candida krusei were observed in 2.3%. Although the distribution of Candida species in other intensive care units and services was comparable to the general incidence, C. albicans was detected in 38%, C. parapsilosis 30% and C. tropicalis 27% in surgical intensive care units. Moreover, C. tropicalis was the dominant species in the neonatal intensive care unit (75%). Conclusions: In the present study, C. albicans was the most common candida species, and C. parapsilosis was the second most frequently reproduced species. It has been suggested that resistance patterns differ between species and between wards, therefore species identification and susceptibility analysis are important, and these should be taken into account when starting empirical, preemptive and antifungal treatment.

Mouth rinses which function as breath fresheners, medicaments, and antiseptics can also deliver oral therapeutic agents. This study evaluated and compared the antifungal effects of synthetic and herbal mouth rinses on oral C. albicans and C. glabrata via disk diffusion, minimal inhibition concentration (MIC), minimum fungicidal concentration (MFC), time-kill assay, and growth profile tests. The four chemical mouth rinses, namely Brand O (A), Brand M (B), Brand H (C), and Brand B (D) used in the study showed positive antifungal activity in these two species. The average diameter of the inhibition zones obtained from the disk diffusion test was higher in mouth rinse B (C. albicans = 12.0 ± 0.9 mm, C. glabrata = 13.5 ± 0.8 mm) compared to those in C, A and D. Both Candida species exhibited similar MIC and MFC values, ranging from 1.63 ± 0.5 to 18.75 ± 0.0 µg/mL and 6.51 ± 2.01 to 50.00 ± 9.36 µg/mL, respectively. These synthetic mouth rinses had efficient killing activity eliminating 50% of the growing population of both Candida spp. following 15 seconds exposure time. Analyses of the growth profile curves showed that mouth rinses B and A resulted in rapid growth depletion of both Candida spp. Meanwhile, three herbal mouth rinses, namely Brand S (E), Brand C (F), and Brand P (G), were less effective against C. albicans and C. glabrata. Mouth rinses B and A contained cetylpyridinium chloride and chlorhexidine, respectively, and could be an effective alternative for controlling and preventing oral candidiasis.

Coronavirus disease 2019 (COVID-19)-associated invasive fungal infections are an important complication in a substantial number of critically ill, hospitalized patients with COVID-19. Three groups of fungal pathogens cause co-infections in COVID-19: Aspergillus, Mucorales and Candida species, including Candida auris. Here we review the incidence of COVID-19-associated invasive fungal infections caused by these fungi in low-, middle- and high-income countries. By evaluating the epidemiology, clinical risk factors, predisposing features of the host environment and immunological mechanisms that underlie the pathogenesis of these co-infections, we set the scene for future research and development of clinical guidance.

This study aimed to evaluate the relationship between oral candidiasis with salivary features, neutrophil function, and glycemic control in type 2 diabetes (T2D).Twenty-nine individuals were included, 16 with T2D and 13 without the disease. The participants underwent clinical examination, neutrophilic function tests, fasting glycemia and glycated hemoglobin (A1c), stimulated and unstimulated saliva collection, and swab and exfoliative cytology. Salivary flow, pH, and total fungi count were evaluated on saliva, and identification of the Candida species was performed in saliva and swab samples.There was no difference in unstimulated salivary flow and pH of the stimulated and unstimulated saliva for participants with T2D and controls (P.05). Individuals from both groups presented no candidal lesions. The salivary fungal growth in the T2D group was higher than that in controls (P.05). Only individuals with T2D presented alterations in the neutrophilic functions (14/16; 87.5%; P.05). There was no relationship between high A1c values and neutrophil dysfunction with the presence of Candida spp. in both saliva and mucosa (P.05).High A1c level, reduction in neutrophil activity, salivary flow and pH, and increase in total salivary Candida spp. counts were not related to oral candidiasis in patients with T2D.

Recent years have witnessed a phenomenal increase in prevalence as well as severity of opportunistic fungal infection (OFIs) despite of availability of different therapeutic options. This could be attributed to various reasons including increased prevalence of immunocomprised patients. Awareness of spectrum of fungal infections in immunocompromised conditions and their epidemiological changes is critical for early diagnosis and proper management. Method: A total of 277 clinical samples were collected from 216 HIV patients suspected of OFI. Sample processing and identification of fungi carried out by standard conventional methods. Result: Out of 216 patients, 30% (65) showed positive fungal etiology with single or multiple system involvement. Oral candidiasis ranked highest (46%) followed by Cryptococcal meningitis (35%), Candidial pneumonia (10%), Oesophageal candidiasis (7%), Dermatophytoses (6%), Pulmonary aspergillosis (4%), Systemic candidiasis (4%), Pneumocystis pneumonia (3%) etc. Discussion: As most of opportunistic fungi (OF) are part of the normal human microbial flora, clinical significance of fungal isolates must be confirmed by standard criteria laid down such as repeated isolation, supportive clinical findings and radiographical evidences. Although Antiretroviral Therapy for minimum 6 months duration provides protection against OFI by maintaining CD4 count, compliance to treatment important.

The incidence of fungal infections particularly Candida species, is increasing gradually as a result of the increased life expectancy associated with the advances in the diagnosis and treatment of diseases, and increased number of patients in the risk group over the years. In addition, the incidence of fungal infection types that are resistant to antifungal drugs has been increasing, and rare fungal species have been reported to be isolated more frequently. For this reason, it is indicated that identification to the species level will contribute to the early initiation of an accurate and effective treatment. In this study, it was aimed to define the Candida species isolated from various clinical specimens and to document the performance of antifungal sensitivity tests. The Candida isolates sent to the central mycology laboratory in 2019 for identification and antifungal susceptibility tests were included in the study. The definition of the fungi to the species level was carried out using matrix-assisted laser desorption ionization-time of fl ight mass spectrometry (MALDI-TOF MS) and conventional methods. In vitro antifungal drug susceptibilities were analyzed using the The Clinical and Laboratory Standarts Institute (CLSI, M27-A3) reference broth microdilution method. The minimum inhibitory concentration (MIC) results were interpreted in accordance with the species-specific clinical breakpoints (CBPs) cited in the CLSI-M60 guidelines, and according to the epidemiological cut-off value (ECV) when no CBP was mentioned. The distribution of the species of the total 813 Candida isolates included in the study were as follows: Candida albicans (n= 312 ), Candida parapsilosis (n= 202), Candida tropicalis (n= 92), Candida glabrata (n= 71), Candida dubliniensis (n=28), Candida lusitaniae (n= 26), Candida kefyr (n= 22), Candida utilis (n= 17), Candida krusei (n= 14), Candida orthopsilosis (n= 7), Candida inconspicua (n= 7), Candida guilliermondii (n= 5), Candida metapsilosis (n= 4), Candida norvegensis (n= 4), Candida lambica (n= 1) and Candida lipolytica (n= 1). The evaluation of the results of the antifungal susceptibility tests according to the CBPs revealed that one C.albicans isolate and 60 C.parapsilosis (29.7%) isolates were resistant, and seven C.parapsilosis (3.5%) isolates were dose-dependent susceptible to fluconazole; 32 C.parapsilosis (15.8%) isolates were intermediately susceptible to voriconazole; one C.parapsilosis (0.5%) was resistant and one C.krusei (7.1%) was intermediately susceptible to anidulafungin; and one C.parapsilosis (0.5%) was resistant and one C.krusei (7.1%) isolate was intermediately susceptible to micafungin. In terms of ECVs, one C.lusitaniae isolate for fluconazole and one of each C.lusitaniae and C.kefyr isolates were evaluated as a non-wild type. In the present study, 61 of 813 isolates were found to be resistant to fluconazole and seven were dose dependently susceptible, 32 were intermediately susceptible to voriconazole, one was resistant to anidulafungin, one was intermediately susceptible, and one was resistant to micafungin and one was intermediately susceptible to micafungin. In conclusion, the increased number of non- albicans Candida species and increased levels of resistance to antifungal drugs further establish the importance of early diagnosis at a species level alongside antifungal susceptibility tests.

Vulvovaginal candidosis (VVC) is a symptomatic vaginal yeast infection, especially caused by Candida spp. Although VVC is common among reproductive-age women, prevalence studies notice the uprise of vaginal Candida colonization to 30% during pregnancy by culture, especially in the last trimester. Recent studies have considered it a severe problem due to the emerging evidence showing the association of VVC with a higher chance of pregnancy-related complexities (e.g., preterm labor, premature rupture of membranes, congenital cutaneous candidosis, and chorioamnionitis). In this review, we have reassessed and summarized the prevalence rate of VVC in expecting mothers and analyzed the association of several factors to the increased risk of VVC during pregnancy in different regions of the world. Altogether, these data collected from various studies showed the highest prevalence of VVC during pregnancy, mostly in Asian and African countries (90.38%, 62.2%, and 61.5% in Kenya, Nigeria, and Yemen, respectively). The prevalence rate of VVC during pregnancy was also found to differ with age, gestation period, parity, educational status, and socioeconomic level. Some pregnancy-related factors (e.g., weakened immunity; elevated level of sex hormones, glycogen deposition; low vaginal pH; decreased cell-mediated immunity) and several clinical and behavioral factors can be suggested as potential risk factors of candidosis during pregnancy.

Approximately 30% of Candida genus isolates are resistant to all currently available antifungal drugs and it is highly important to develop new treatments. Additionally, many current drugs are toxic and cause unwanted side effects. 1,3-beta-glucan synthase is an essential enzyme that builds the cell walls of Candida.Targeting CaFKS1, a subunit of the synthase, could be used to fight Candida.In the present study, a machine-learning model based on chemical descriptors was trained to recognise drugs that inhibit CaFKS1. The model attained 96.72% accuracy for classifying between active and inactive drug compounds. Descriptors for FDA-approved and other drugs were calculated, and the model was used to predict the potential activity of these drugs against CaFKS1.Several drugs, including goserelin and icatibant, were detected as active with high confidence. Many of the drugs, interestingly, were gonadotrophin-releasing hormone (GnRH) antagonists or agonists. A literature search found that five of the predicted drugs inhibit Candida experimentally.This study yields promising drugs to be repurposed to combat Candida albicans infection. Future steps include testing the drugs on fungal cells in vitro.

Infections due to Candida species represent a serious threat to healthcare facilities. Only a few classes of antifungal agents are available, and the rapid emergence of (multi)drug resistance eliminates effective treatment options for successful patient outcome. Topically applied antiseptics may represent a suitable tool for infection control and local therapy. This study aimed to investigate the in vitro efficacy of the widely used antiseptic octenidine (OCT) against clinical isolates of emerging azole-, echinocandin- and multi-resistant Candida albicans and Candida glabrata.The antifungal activity of different concentrations of OCT ranging from 0.001% to 0.05% and of OCT-containing ready-to-use products was determined against well-characterised (multidrug) resistant C. albicans and C. glabrata isolates, including susceptible wild-type strains. Quantitative suspension tests were performed under "clean conditions" (0.3 g/L bovine serum albumin) and under "dirty conditions" (3 g/L albumin + 3 mL/L defibrinated sheep blood) as well as various contact times (30 s, 1 min, 2 min) according to EN13624:2013.Even in the presence of a high organic load, pure OCT at 0.05% and a contact time of 30 s was fully effective for all Candida strains, with growth kinetics indicating a time- and concentration-dependent activity. Importantly, commercially available OCT-based products achieved the required reduction of ≥4 logThese results encourage consideration of the well-tolerated antiseptic molecule OCT in the eradication of emerging (multidrug) resistant C. albicans and C. glabrata.

Lignocellulose is an abundant xylose-containing biomass found in agricultural wastes, and has arisen as a suitable alternative to fossil fuels for the production of bioethanol. Although Saccharomyces cerevisiae has been thoroughly used for the production of bioethanol, its potential to utilize lignocellulose remains poorly understood. In this work, xylose-metabolic genes of Pichia stipitis and Candida tropicalis, under the control of different promoters, were introduced into S. cerevisiae. RNA-seq analysis was use to examine the response of S. cerevisiae metabolism to the introduction of xylose-metabolic genes. The use of the PGK1 promoter to drive xylitol dehydrogenase (XDH) expression, instead of the TEF1 promoter, improved xylose utilization in ?XR-pXDH? strain by overexpressing xylose reductase (XR) and XDH from C. tropicalis, enhancing the production of xylitol (13.66 ? 0.54 g/L after 6 days fermentation). Overexpression of xylulokinase and XR/XDH from P. stipitis remarkably decreased xylitol accumulation (1.13 ? 0.06 g/L and 0.89 ? 0.04 g/L xylitol, respectively) and increased ethanol production (196.14% and 148.50% increases during the xylose utilization stage, respectively), in comparison with the results of XR-pXDH. This result may be produced due to the enhanced xylose transport, Embden?Meyerhof and pentose phosphate pathways, as well as alleviated oxidative stress. The low xylose consumption rate in these recombinant strains comparing with P. stipitis and C. tropicalis may be explained by the insufficient supplementation of NADPH and NAD+. The results obtained in this work provide new insights on the potential utilization of xylose using bioengineered S. cerevisiae strains.

Each year, millions of central venous catheter insertions are performed in intensive care units worldwide. The usage of these indwelling devices is associated with a high risk of bacterial and fungal colonization, leading to the development of microbial consortia, namely biofilms. These sessile structures provide fungal cells with resistance to the majority of antifungals, environmental stress and host immune responses. Based on different guidelines, colonized/infected catheters should be removed and changed immediately in the case of Candida-related central line infections. However, catheter replacement is not feasible for all patient populations. An alternative therapeutic approach may be antifungal lock therapy, which has received high interest, especially in the last decade. This review summarizes the published Candida-related in vitro, in vivo data and case studies in terms of antifungal lock therapy. The number of clinical studies remains limited and further studies are needed for safe implementation of the antifungal lock therapy into clinical practice.

The peptide transport (PTR) or proton-dependent oligopeptide transporter (POT) family exploits the inwardly directed proton motive force to facilitate the cellular uptake of di/tripeptides. Interestingly, some representatives are also shown to import peptide-based antifungals in certain Candida species. Thus, the identification and characterization of PTR transporters serve as an essential first step for their potential usage as antifungal peptide uptake systems. Herein, we present a genome-wide inventory of the PTR transporters in five prominent Candida species. Our study identifies 2 PTR transporters each in C. albicans and C. dubliniensis, 1 in C. glabrata, 4 in C. parapsilosis, and 3 in C. auris. Notably, despite all representatives retaining the conserved features seen in the PTR family, there exist two distinct classes of PTR transporters that differ in terms of their sequence identities and lengths of certain extracellular and intracellular segments. Further, we also evaluated the contribution of each PTR protein of the newly emerged multi-drug-resistant C. auris in di/tripeptide uptake. Notably, deletion of two PTR genes BNJ08_003830 and BNJ08_005124 led to a marked reduction in the transport capabilities of several tested di/tripeptides. However, all three genes could complement the role of native PTR2 gene of Saccharomyces cerevisiae, albeit to varied levels. Besides, BNJ08_005124 deletion also resulted in increased resistance toward the peptide-nucleoside drug Nikkomycin Z as well as the glucosamine-6-phosphate synthase inhibitor, L-norvalyl-N3-(4-methoxyfumaroyl)-L-2,3-diaminopropionoic acid (Nva-FMDP), pointing toward its predominant role in their uptake mechanism. Altogether, the study provides an important template for future structure-function investigations of PTR transporters in Candida species. KEY POINTS: • Candida genome encodes for two distinct classes of PTR transporters. • Candida auris encodes for 3 PTR transporters with different specificities. • BNJ08_005124 in C. auris is involved in the uptake of Nikkomycin Z and Nva-FMDP.

Aims This study aimed to evaluate the combined effect of a mannose-binding lectin Helja with fluconazole (FLC) on Candida albicans and to get insights about the joint action mechanism. Methods and Results The fungal growth was assessed following the optical density at 630 nm. Fungal cell morphology and nucleus integrity were analysed by flow cytometry and confocal laser scanning microscopy using Calcofluor White (CFW) and 4′,6-diamidino-2-phenylindole (DAPI) staining respectively. The basis of Helja + FLC action on cell wall and plasma membrane was analysed using perturbing agents. The Helja + FLC combination exhibited an inhibitory effect of fungal growth about three times greater than the sum of both compounds separately and inhibited fungal morphological plasticity, an important virulence attribute associated with drug resistance. Cells treated with Helja + FLC showed morphological changes, nucleus disintegration and formation of multimera structures, leading to cell collapse. Conclusions Our findings indicate that the Helja + FLC combination exhibited a potent antifungal activity based on their simultaneous action on different microbial cell targets. Significance and Impact of Study The combination of a natural protein with conventional drugs might be helpful for the design of effective therapeutic strategies against Candida, contributing to minimize the development of drug resistance and host cell toxicity.

Background: Candida is an opportunistic fungus often present in the oral mucosa. In the compromised immune system, it may become pathogenic and cause oral candidiasis. This infection is more common with Candida albicans; though, non-albicans Candida spp also have significant relevance. Current treatment guidelines include polyenes, azoles and echinocandins, where fluconazole is the primary therapeutic option. However, both inherited and acquired resistance to fluconazole is exhaustively reported. The development of resistance has resulted in the worsening of the original and re-emergence of new fungal diseases. Thus, the development of an anti-candidiasis therapy with a satisfactory outcome is the urgent need of the hour. Objective: This review article aims to stimulate research in establishing the synergistic efficacy of various flavonoids with fluconazole to combat the resistance and develop an effective pharmacotherapy for the treatment of oral candidiasis. Further, in this article, we discuss in detail the mechanisms of action of fluconazole, along with the molecular basis of the development of resistance in Candida species. Method: PubMed and other databases were used for literature search. Results: The designing of natural drugs from the plant-derived phytochemicals are the promising alternatives in modern medicine. The challenge today is the development of alternative anti-oral candidiasis drugs with increased efficacy, bioavailability and better outcome which can combat azole resistance. Identifying the flavonoids with potential antifungal action at low concentrations seems to meet the challenges. Conclusion: Phyto-active constituents, either alone or in combination with conventional antibiotics may be an effective approach to deal with global antimicrobial resistance. The efficacy of herbal therapy for decades suggests that bacteria, fungi, and viruses may have a reduced ability to adapt and resistance to these natural antimicrobial regimes.

Candidemia is rare and has a high mortality rate. This study analyses the impact of bedside antifungal stewardship (AFS) on clinical management and prognosis of patients with candidemia at a university hospital in Germany.All patients with at least one positive blood culture with Candida species between 2014 and 2016 received bedside AFS with standardized recommendations. Medical records were retrospectively analyzed. Results from the intervention period from 2014-2016 (n=109), with focus on 2016 (n=39), were compared with those from the pre-intervention period in 2013 (n=30).Bedside AFS was performed in 24/35 (69%) surviving patients in 2016 within the first 3 days after diagnosis of candidemia. All surviving patients (n=35) in 2016 received antifungal treatment compared with 24/28 (86%) in 2013 (p=0.0344). Follow-up blood cultures were performed in 25/35 (71%) in 2016 compared with 10/25 (40%) in 2013 (p=0.0046). Survival in the intervention compared with the pre-intervention group did not differ significantly (p=0.58) one year after the diagnosis of candidemia was made. However, patients with candidemia often have multiple serious comorbidities.Individualized bedside AFS significantly improves adherence to recommendations for patients with Candida fungemia, especially guideline-oriented diagnostics and therapy. Improving the prognosis of patients with candidemia remains a huge challenge for AFS.

Background Hybrids are chimeric organisms with highly plastic heterozygous genomes that may confer unique traits enabling the adaptation to new environments. However, most evolutionary theory frameworks predict that the high levels of genetic heterozygosity present in hybrids from divergent parents are likely to result in numerous deleterious epistatic interactions. Under this scenario, selection is expected to favor recombination events resulting in loss of heterozygosity (LOH) affecting genes involved in such negative interactions. Nevertheless, it is so far unknown whether this phenomenon actually drives genomic evolution in natural populations of hybrids. To determine the balance between selection and drift in the evolution of LOH patterns in natural yeast hybrids, we analyzed the genomic sequences from fifty-five hybrid strains of the pathogenic yeasts Candida orthopsilosis and Candida metapsilosis, which derived from at least six distinct natural hybridization events. Results We found that, although LOH patterns in independent hybrid clades share some level of convergence that would not be expected from random occurrence, there is an apparent lack of strong functional selection. Moreover, while mitosis is associated with a limited number of inter-homeologous chromosome recombinations in these genomes, induced DNA breaks seem to increase the LOH rate. We also found that LOH does not accumulate linearly with time in these hybrids. Furthermore, some C. orthopsilosis hybrids present LOH patterns compatible with footprints of meiotic recombination. These meiotic-like patterns are at odds with a lack of evidence of sexual recombination and with our inability to experimentally induce sporulation in these hybrids. Conclusions Our results suggest that genetic drift is the prevailing force shaping LOH patterns in these hybrid genomes. Moreover, the observed LOH patterns suggest that these are likely not the result of continuous accumulation of sporadic events—as expected by mitotic repair of rare chromosomal breaks—but rather of acute episodes involving many LOH events in a short period of time. This work was supported by the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No. H2020-MSCA-ITN-2014-642095. The TG group also acknowledges the support from the Spanish Ministry of Economy, Industry, and Competitiveness (MEIC) for the EMBL partnership and grants “Centro de Excelencia Severo Ochoa 2013-2017” SEV-2012-0208 and BFU2015-67107 co-founded by the European Regional Development Fund (ERDF); from the CERCA Programme/Generalitat de Catalunya; from the Catalan Research Agency (AGAUR) SGR857 and grants from the European Union’s Horizon 2020 research and innovation program under the grant agreement ERC-2016-724173. TG also receives support from an INB Grant (PT17/0009/0023—ISCIII-SGEFI/ERDF). The authors thank Dr. Powel Golik’s guidance in the identification of PPR proteins, Simone Mozzachiodi and Dr. Gianni Liti for the helpful discussions on the analysis of meiotic patterns, and all Gabaldón lab members for the helpful discussions and comments on this work, especially Marina Marcet-Houben.

The opportunistic fungal infections are an increasing threat to humans due to the increasing number of patients with immunodeficiency, in which the most popular fungal pathogen is Candida albicans. Fluconazole (FLC) is the common drug for treating C. albicans infections, but increasing drug resistance has limited its clinical use. Currently, combination therapy is being investigated as a treatment to overcome the resistance of C. albicans. This report investigated the synergistic properties of deferoxamine (DFO) and FLC combination therapy in vitro and in vivo against drug-resistant C. albicans. The results showed that the combination of DFO and FLC had a great synergistic antifungal effect against C. albicans, an FLC-resistant strain, with a fractional inhibition concentration index (FICI) of 0.25 by the broth microdilution checkerboard assay. Furthermore, the combination of DFO and FLC significantly inhibited the activity of C. glabrata cells (approximately 30% of C. glabrata cells are azole-resistant). The time-growth curves confirmed that the combination of DFO and FLC have a potent synergistic antifungal effect. Hyphal formation assays confirmed that DFO inhibited the hyphal induction of C. albicans. In addition, the combination of DFO and FLC significantly inhibited the expression of the adhesion gene (ALS1). In vivo experiments showed that the combination of DFO and FLC significantly reduced pustules, CFU counts and inflammatory cell infiltration in skin tissue. These results suggest that the combination of DFO and FLC inhibits yeast-hyphae transformation, reduces C. albicans infectivity and resistance in vitro and in vivo, and affects Cek1 MAPK signaling. This may offer a new option for the treatment of cutaneous candidiasis.

Biofilm and hypha formation are central to virulence of the fungal pathogen Candida albicans. The G1 cyclin gene HGC1 is required for hypha formation under diverse in vitro and in vivo growth conditions. Hgc1 is required for disseminated infection and is a linchpin in the argument that hyphal morphogenesis itself is required for pathogenicity. We report here that HGC1 is dispensable for hypha formation during biofilm formation both in vitro, under strong inducing conditions, and in vivo, in a mouse oropharyngeal candidiasis model. These findings are validated with two or more C. albicans isolates. Systematic screening of overexpressed cyclin genes indicates that CCN1 and CLN3 can compensate partially for Hgc1 function during biofilm growth. This conclusion is also supported by the severity of the hgc1Δ/Δ ccn1Δ/Δ double mutant biofilm defect. Our results suggest that hypha formation in biofilm is accomplished by combined action of multiple cyclins, not solely by Hgc1. IMPORTANCE The HGC1 gene encodes a cyclin that is required for virulence of the fungal pathogen Candida albicans. It is required to produce the elongated hyphal filaments of free-living planktonic cells that are associated with virulence. Here, we show that HGC1 is not required to produce hyphae in the alternative growth form of a biofilm community. We observe Hgc1-independent hyphae in two infection-relevant situations, biofilm growth in vitro and biofilm-like oropharyngeal infection. Our analysis suggests that hypha formation in the biofilm state reflects combined action of multiple cyclins.

In this issue of Cell Hostamp; Microbe, Basso et al. delineate a mechanism for the fungal lipase Lip2 during systemic infection with Candida albicans, the most common human fungal pathogen. They find that lipids liberated by Lip2 can manipulate the protective host cytokine response, thereby enhancing fungal virulence.

Candida auris is an emerging multidrug-resistant pathogen in intensive care settings (ICU). During the coronavirus disease 19 (COVID-19) pandemic, ICU admissions were overwhelmed, possibly contributing to the C. auris outbreak in COVID-19 patients.The present systematic review addresses the prevalence, underlying diseases, iatrogenic risk factors, treatment and outcome of C. auris infections in COVID-19 patients.MEDLINE, Scopus, Embase, Web of Science and LitCovid databases were systematically searched with appropriate keywords from 1 January 2020 to 31 December 2021.A total of 97 cases of C. auris were identified in COVID-19 patients. The pooled prevalence of C. auris infections (encompassing candidemia and non-candidemia cases) in COVID-19 patients was 14%. The major underlying diseases were diabetes mellitus (42.7%), hypertension (32.9%) and obesity (14.6%), followed by the iatrogenic risk factors such as a central venous catheter (76.8%%), intensive care unit (ICU) stay (75.6%) and broad-spectrum antibiotic usage (74.3%). There were no significant differences in underlying disease and iatrogenic risk factors among C. auris non-candidemia/colonisation and C. auris candidemia cases. The mortality rate of the total cohort is 44.4%, whereas, in C. auris candidemia patients, the mortality was 64.7%.This study shows that the prevalence of C. auris infections remains unchanged in the COVID-19 pandemic. Hospital-acquired risk factors may contribute to the clinical illness. Proper infection control practices and hospital surveillance may stop future hospital outbreaks during the pandemic.

O abscesso cerebral por Candida spp. é uma condição ameaçadora à vida, mais comum em indivíduos imunocomprometidos, porém com relatos crescentes em pacientes imunocompetentes. Esta revisão sistemática objetiva avaliar os principais fatores de risco, para além de distúrbios imunológicos, relacionados ao abscesso cerebral por Candida, bem como os métodos diagnósticos e tratamentos recomendados. Foram utilizados os descritores DeCS “Brain abscess” e “Candida” para uma pesquisa na base de dados MEDLINE, restrita aos trabalhos publicados até dezembro de 2021, encontrando-se um total de 88 publicações; destes, 48 atenderam aos critérios de inclusão deste estudo. Observou-se que a idade média dos pacientes foi de 21 anos, com maior prevalência no sexo masculino (56%). Prematuridade (57%) e baixo peso ao nascer (57%), em crianças, além de uso prévio de antibióticos de amplo espectro (59%) e infecção prévia por Candida (47%), nos pacientes em geral, foram os principais fatores de risco identificados, enquanto 6% não apresentaram qualquer fator de risco. Alteração do nível de consciência (67%) e febre (47%) foram as principais manifestações clínicas e o diagnóstico foi realizado em grande parte por biópsia (65%). Candida albicans foi a espécie mais frequentemente envolvida (93%). A terapia antifúngica foi variável, sendo precedida por antibioticoterapia em 53% dos casos. A taxa de mortalidade foi de 24%. Houve retardo no início da terapia antifúngica na maioria dos casos devido à dificuldade diagnóstica, evidenciado a necessidade de alto nível de suspeição e métodos diagnósticos sensíveis para otimização da terapêutica e desfecho dos casos, com redução das taxas de mortalidade.

Lipase B from Candida antarctica (CALB) and lipases from Candida rugosa (CRL) and Rhizomucor miehei (RML) have been coimmobilized on octyl and octyl-Asp agarose beads. CALB was much more stable than CRL, that was significantly more stable than RML. This forces the user to discard immobilized CALB and CRL when only RML has been inactivated, or immobilized CALB when CRL have been inactivated. To solve this problem, a new strategy has been proposed using three different immobilization protocols. CALB was covalently immobilized on octyl-vinyl sulfone agarose and blocked with Asp. Then, CRL was immobilized via interfacial activation. After coating both immobilized enzymes with polyethylenimine, RML could be immobilized via ion exchange. That way, by incubating in ammonium sulfate solutions, inactivated RML could be released enabling the reuse of coimmobilized CRL and CALB to build a new combi-lipase. Incubating in triton and ammonium sulfate solutions, it was possible to release inactivated CRL and RML, enabling the reuse of immobilized CALB when CRL was inactivated. These cycles could be repeated for 3 full cycles, maintaining the activity of the active and immobilized enzymes.

Although oxygen concentrations inside of the human body vary depending on organs or tissues, few reports describe the relationships between biofilm formation of Candida species and oxygen concentrations. In this study, we investigated the biofilm-forming capabilities of Candida species under various oxygen conditions.We evaluated the adhesion and biofilm formation of Candida albicans and C. tropicalis under aerobic, microaerobic (oxygen concentration 5%), or anaerobic conditions. We also examined how oxygen concentration affects adhesion/maturation by changing adhesion/maturation phase conditions. We used crystal violet assay to estimate the approximate biofilm size, performed microscopic observation of biofilm morphology, and evaluated adhesion-associated gene expression.The adhered amount was relatively small except for a clinical strain of C. tropicalis. Our biofilm-formation analysis showed that C. albicans formed a higher-size biofilm under aerobic conditions, while C. tropicalis favored microaerobic conditions to form mature biofilms. Our microscopic observations were consistent with these biofilm-formation analysis results. In particular, C. tropicalis exhibited more hyphal formation under microaerobic conditions. By changing the adhesion/maturation phase conditions, we represented that C. albicans had favorable biofilm-formation capability under aerobic conditions, while C. tropicalis showed enhanced biofilm formation under microaerobic adhesion conditions. In good agreement with these results, the C. tropicalis adhesion-associated gene expression tended to be higher under microaerobic or anaerobic conditions.C. albicans favored aerobic conditions to form biofilms, whereas C. tropicalis showed higher biofilm-formation ability and promoted hyphal growth under microaerobic conditions. These results indicate that favorable oxygen conditions significantly differ for each Candida species.

Candida auris a frequently multidrug-resistant yeast species that poses a global health threat due to its high potential for hospital outbreaks. While C. auris has become endemic in parts of Asia and Africa, transmissions have so far rarely been reported in Western Europe except for Great Britain and Spain. We describe the first documented patient-to-patient transmission of C. auris in Germany in a COVID-19 intensive care unit (ICU) and infection control measures implemented to prevent further spread of the pathogen.Identification of C. auris was performed by MALDI-TOF and confirmed by internal transcribed spacer (ITS) sequencing. Antifungal susceptibility testing was carried out. We conducted repeated cross-sectional examinations for the presence of C. auris in the patients of the affected ICU and investigated possible routes of transmission.The index patient had been transferred to Germany from a hospital in Northern Africa and was found to be colonised with C. auris. The contact patient developed C. auris sepsis. Infection prevention and control (IPC) measures included strict isolation of the two C. auris patients and regular screening of non-affected patients. No further case occurred during the subsequent weeks. Reusable blades used in video laryngoscope-guided intubation were considered as the most likely vehicle of transmission.In view of its high risk of transmission, vigilance regarding C. auris colonisation in patients referred from endemic countries is crucial. Strict and immediate IPC measures may have the potential to prevent C. auris outbreaks.

The COVID-19 pandemic has intensified interest in how the infection affects the lung microbiome of critically ill patients and how it contributes to acute respiratory distress syndrome (ARDS). We aimed to characterize the lower respiratory tract mycobiome of critically ill patients with COVID-19 in comparison to patients without COVID-19.We performed an internal transcribed spacer 2 (ITS2) profiling with the Illumina MiSeq platform on 26 respiratory specimens from patients with COVID-19 as well as from 26 patients with non-COVID-19 pneumonia.Patients with COVID-19 were more likely to be colonized with Candida spp. ARDS was associated with lung dysbiosis characterized by a shift to Candida species colonization and a decrease of fungal diversity. We also observed higher bacterial phylogenetic distance among taxa in colonized patients with COVID-19. In patients with COVID-19 not colonized with Candida spp., ITS2 amplicon sequencing revealed an increase of Ascomycota unassigned spp. and 1 Aspergillus spp.-positive specimen. In addition, we found that corticosteroid therapy was frequently associated with positive Galactomannan cell wall component of Aspergillus spp. among patients with COVID-19.Our study underpins that ARDS in patients with COVID-19 is associated with lung dysbiosis and that an increased density of Ascomycota unassigned spp. is present in patients not colonized with Candida spp.

Aims To assess the efficacy of the antifungal, occidiofungin, against Candida albicans and Candida tropicalis morphological transformation. Methods and Results Susceptibility assays and morphological data were used to demonstrate that occidiofungin effectively targets C. albicans and C. tropicalis undergoing morphological transformation. Susceptibility assays found that cell sensitivity to occidiofungin varied with the media conditions used for morphological switching. Microscopy data showed that occidiofungin inhibited hyphae formation when added at the time of morphological induction and hyphal extension when added within the first hour following hyphae induction. Immunoblot analysis demonstrated that occidiofungin addition prevented activation of Cek1p MAPK signalling. Conclusions The data indicated that the antimicrobial compound, occidiofungin, effectively targets hyphae elongation in Candida spp. and suggests the biological target of occidiofungin is necessary for the morphological changes associated with yeast-to-hyphae switching. Significance and Impact of the Study Findings from this study demonstrated that occidiofungin effectively targets the invasive growth of dimorphic Candida which suggests this compound may also inhibit the heterogenous population of cells present in a clinical setting. This presents occidiofungin as a promising candidate for the treatment of Candida associated infections.

The yeasts involved in the ripening process of artisanal soft raw ewe milk Protected Designation of Origin (PDO) Torta del Casar and Queso de la Serena cheeses produced in Extremadura, Spain, were isolated throughout their ripening process, strain typed, and characterized for some important technological properties. A total of 508 yeast isolates were obtained and identified by inter-single sequence repeat anchored PCR amplification analysis and subsequent sequencing of the internal transcribed spacer ITS1/ITS2 5.8S rRNA. A total of 19 yeast species representing 8 genera were identified. Debaryomyces hansenii, Pichia kudriavzevii, Kluyveromyces lactis, and Yarrowia lipolytica were the predominant species. We selected 157 isolates, by genotyping and origin, for technological characterization. The evaluation of yeast isolates' growth under stress conditions of cheese ripening showed that 87 presented better performance. Among them, 71 isolates were not able to catabolize tyrosine to produce a brown pigment. Principal component analysis of the biochemical features of these isolates showed that 9 strains stood out, 3 K. lactis strains (2287, 2725, and 1507), 2 Pichia jadinii (1731 and 433), 2 Yarrowia alimentaria (1204 and 2150), Y. lipolytica 2495 and P. kudriavzevii 373. These strains displayed strong extracellular proteolytic activity on skim milk agar as well as an adequate enzymatic profile (strong aminopeptidase and weak protease activity), suggesting their great potential for cheese proteolysis. Extracellular lipolytic activity was mainly restricted to Yarrowia spp. isolates and weakly present in P. kudriavzevii 373 and K. lactis 2725, although enzymatic characterization by API-ZYM (bioMérieux SA) evidenced that all may contribute, at least in part, to the lipolysis process. Moreover, these strains were able to assimilate lactose, galactose, and glucose at NaCl concentrations higher than that usually found in cheese. However, lactate and citrate assimilation were limited to Y. lipolytica 2495, P. kudriavzevii 373, and P. jadinii 433, and may contribute to the alkalinizing process relevant to biochemical processes that take place in the last stages of ripening. By contrast, K. lactis strains showed acidifying capacity and β-galactosidase activity and may take part in the initial stages of ripening, together with lactic acid bacteria. Thus, considering the technological characteristics studied, the 9 selected strains presented biochemical features well suited to their potential use as adjunct cultures, alone or in combination with autochthonous starter bacteria in the cheesemaking process, to overcome the heterogeneity of these PDO cheeses, preserving their unique sensory characteristics.

Invasive fungal infection, particularly intraabdominal candidiasis, exerts a negative impact on the outcome of pancreas transplant recipients (PTRs). Optimal antifungal prophylaxis in this context remains unclear.We performed a single-centre retrospective study to compare the incidence of invasive candidiasis during the first 6 post-transplant months in a cohort of 218 PTRs over two periods in which different agents for antifungal prophylaxis were used: fluconazole (Fluco-Px) from March 1995 to June 2012, and micafungin followed by fluconazole (Mica-Px) from July 2012 to December 2018.A total of 152 and 66 PTRs received Fluco-Px and Mica-Px. Mean age was 39.7 ± 7.8 years, 56.4% (123/218) were males, and 85.3% (186/218) underwent simultaneous pancreas-kidney transplantation. Invasive candidiasis occurred in 21.7% (33/152) of PTRs under Fluco-Px compared to 24.2% (16/66) of those under Mica-Px (p-value = .681). Median time from transplantation to infection was 8 days (interquartile range [IQR]: 6-16) under Fluco-Px versus 6.5 (IQR: 3.3-15.8) under Mica-Px (p-value = .623). Non-albicans Candida species comprised 27.5% (11/40) and 25.0% (4/16) of episodes under Fluco-Px and Mica-Px respectively (p-value = .849). Surgical site infection was the most common form in both groups (82.5% [33/40] and 87.5% [14/16]; p-value = .954). Multivariable analysis identified cold ischaemia time of the pancreas and kidney grafts, surgical reintervention and insulin requirement after transplantation as risks factor for invasive candidiasis.This retrospective study did not reveal a significant benefit from the initial use of micafungin-based antifungal prophylaxis over fluconazole among PTRs in terms of invasive candidiasis.

Introduction: Candidemia causes high mortality and is occuring at increasing rate in intensive care units (ICUs). (1,3)- β-D-glucan (BDG) testing is recommended in neutropenic patients. However the usefulness of BDG in ICUs is unclear. Methodology: This study was conducted to compare the diagnostic value of Candida score (CS), colonization index (CI), serum BDG detection, and routine laboratory parameters in ICU patients. Characteristics and laboratory data of 83 patients (15 patients with candidemia and 68 patients without candidemia) were evaluated. Results: Median serum BDG was significantly higher in the candidemia group (129 pg/mL vs. 36 pg/mL, p < 0.001). BDG assay with standard cut-off value ≥ of 80 pg/mL had 93.33% sensitivity and 64.18% specificity (Areas under the ROC curve (AUC): 0.788). This study concluded that the optimal cut-off value for BDG assay was 112 pg/mL with sensitivity of 86.67% and specificity of 82.09% (AUC: 0.844). C-reactive protein (CRP) with optimal cut-off value ≥ 85 mg/L and BDG ≥ 80 pg/mL had the highest AUC (0.862, 95% CI: 0.768 - 0.928) with sensitivity 93.33% and specificity 79.1%. Conclusions: Predicting candidemia is essential in critically ill patients who are at high risk and have high mortality rates. The results of this study suggest that BDG testing is useful for predicting candidemia in ICU. However, BDG combined with CRP may be a stronger predictor for candidemia.

Under natural environmental settings or in the human body, the majority of microorganisms exist in complex polymicrobial biofilms adhered to abiotic and biotic surfaces. These microorganisms exhibit symbiotic, mutualistic, synergistic, or antagonistic relationships with other species during biofilm colonization and development. These polymicrobial interactions are heterogeneous, complex, and hard to control, thereby often yielding worse outcomes than monospecies infections. Concerning fungi, Candida spp., in particular, Candida albicans is often detected with various bacterial species in oral biofilms. These Candida-bacterial interactions may induce the transition of C. albicans from commensal to pathobiont or dysbiotic organism. Consequently, Candida–bacterial interactions are largely associated with various oral diseases, including denture stomatitis, dental caries, periodontitis, peri-implantitis, endodontic infections, and oral cancer. Given the severity of oral diseases caused by cross-kingdom consortia that develop hard-to-remove and highly drug-resistant biofilms, fundamental research is warranted to strategically develop cost-effective and safe therapies to prevent and treat cross-kingdom interactions and subsequent biofilm development. While studies have shed some light, targeting fungal-involved polymicrobial biofilms has been limited. This mini-review outlines the key features of Candida-bacterial interactions and their impact on various oral diseases. In addition, current knowledge on therapeutic strategies to target Candida-bacterial polymicrobial biofilms is discussed.

Bloodstream infection (BSI) by species of Candida has been identified as an important cause of death in patients with neutropenia who undergo chemotherapy for the treatment of hematologic malignancies This study aimed to verify the occurrence of bloodstream infections by Candida species in patients admitted to the haematology-oncology service of a public hospital specialized in the treatment of cancer in Northeast Brazil. A total of 105 clinical samples from 62 patients with haematological malignancies were analyzed at the Laboratory of Medical Mycology at the Federal University of Pernambuco. Only 7 of 105 individuals were in the ICU environment. The mycological diagnosis was performed through automation (BACTEC 9120 / PHOENIX™), proteomic identification (MALDI-TOF MS) and molecular analysis (PCR). The antifungal susceptibility test followed the bloodstream infection recommendations of Clinical Laboratory Standards Institute. Among the samples studied, nine strains (8,57%) were of the genus Candida, being six C. tropicalis and three C. albicans. The isolates were completely susceptible to the antifungal agents tested. Deaths occurred in 66,6% of the cases. Patients with hematologic malignancies hospitalized in intensive care and the state of septic shock present a higher risk of occurrence of BSI by Candida and death by this opportunistic pathogen.

Candida tropicalis is the most common non-albicans Candida species found in Asia-Pacific countries, including Thailand. The pathogen is known for its great virulence, which causes a high case-fatality rate. Associations between case fatality and patient characteristics, infectious disease unit consultation and EQUAL Candida score were investigated.This retrospective cohort study was conducted with 160 cases of C. tropicalis bloodstream infection between 2015 and 2019 at a single, large, tertiary centre in Thailand. Clinical characteristics, clinical presentations, patient outcomes (30-day case-fatality rate) and independent predictive factors were analysed.The 30-day case-fatality rate was 68.1%. The median of the EQUAL Candida score was 8. Independent factors for the prediction of case fatality were septic shock (hazard ratio, 1.84), the use of mechanical ventilation (hazard ratio, 2.03) and the EQUAL Candida score (hazard ratio, 0.75).The predictive factors for 30-day case fatality were septic shock, mechanical ventilation use and the EQUAL Candida score. It is recommended that the EQUAL score be considered for patients infected with C. tropicalis candidaemia to reduce the case-fatality rate.

The rise of antimicrobial-resistant phenotypes and the spread of the global pandemic of COVID-19 are worsening the outcomes of hospitalized patients for invasive fungal infections. Among them, candidiases are seriously worrying, especially since the currently available drug armamentarium is extremely limited. We recently reported a new class of macrocyclic amidinoureas bearing a guanidino tail as promising antifungal agents. Herein, we present the design and synthesis of a focused library of seven derivatives of macrocyclic amidinoureas, bearing a second phenyl ring fused with the core. Biological activity evaluation shows an interesting antifungal profile for some compounds, resulting to be active on a large panel of Candida spp. and C. neoformans. PAMPA experiments for representative compounds of the series revealed a low passive diffusion, suggesting a membrane-based mechanism of action or the involvement of active transport systems. Also, compounds were found not toxic at high concentrations, as assessed through MTT assays.

Introduction: Vaginitis is one of the most common complaints of women who present to health institutions. This study aimed to evaluate the results of vaginal culture obtained from patients who were admitted to the outpatient clinic because of recurrent vaginitis. Methods: Patient records including vaginal culture results of patients aged 18-49 years who were admitted to the gynecology outpatient clinics of the University of Health Sciences Turkey, İstanbul Training and Research Hospital between January 2018 and June 2020 with the complaint of recurrent vaginitis were analyzed retrospectively. Results: The mean age of the 206 patients included in the study was 34.4±12.2 years. Of the 206 vaginal cultures, 124 (60,1%) were negative and 82 (39.8%) were culture positive. Thirty-four patients (16,5%) had mixt microorganisms, 22 had Candida species (10,6%) and 11 had Gardnerella vaginalis (5,3%). Various bacteria were isolated in 15 patients. Conclusion: The rate of vaginitis recurrence and type of microorganism detected can be affected by various factors such as sociocultural structure, race, education level, and contraception method. Only a few studies examined vaginal culture results in patients with recurrent and persistent vaginitis in general and in Turkey. In this study, the contribution of vaginal culture to patient management was limited in this patient group; moreover, the incidence of recurrent vaginitis was lower in women who were using the barrier method.

SUMMARY Backgoround Candida auris infections were reported worldwide since the pathogen was isolated in 2009. In a hospital setting of a C. auris epidemic, we analyzed incidence of cutaneous and intestinal colonization, and connection with infections by the organism. Methods This was a retrospective study in intensive care units at a 1200-bed Italian hospital. The incidence of cutaneous positive swabs, and cutaneous carriers, for C. auris was compared to that of rectal positive swabs, and intestinal carriers, and both were correlated to C. auris infections. Findings A total of 399 patients were included. Seventy-seven patients were infected by C. auris. Ratio of C. auris positive skin swabs from screening in ICUs was 24%. The ratio obtained from infected patients and intestinal C. auris carriers was 49.1%, likewise rectal swabs from similar cohort of patients (P = 0.373). 39.7% and 5.5% of this cohort was colonized only in skin and in rectum, respectively, while 54.8% was colonized in both sites. 12.3% and 83.6% of skin swabs was, respectively, always positive and variable over time in single subject, while 31.5% and 41.1% of rectal swabs were always positive and variable (P = 0.000). Intestinal colonization was associated with increased risk for C. auris urinary infections (P = 0.006). Conclusions C. auris intestinal carriers were fewer than cutaneous carriers, but more continously colonized. Rectal and skin swabs can be good tools for surveillance, respectively, of colonization and of hygiene measures effectiveness. Urinary tract infections by C. auris appeared increasing with gastrointestinal presence of the yeast.