In recent years, Candida glabrata (C. glabrata) has emerged as a pathogen responsible for systemic mortal infections. C. glabrata invades nonphagocytic cells, but the mechanisms involved in its internalization and its intracellular fate in these cells remain poorly understood. Here, it was shown that endocytosis of C. glabrata in epithelial cells partially depends on actin and microtubule rearrangements; importantly, C. glabrata promotes its uptake. The analysis of intracellular fate determined that C. glabrata avoids the fusion of endocytic vacuoles with lysosomes and replicates in epithelial cells. Additionally, C. glabrata downregulates host cell autophagy in the first hour of infection, which correlates with its intracellular replication. Remarkably, the ectopic activation of autophagy contributed to the control of intracellular growth of this yeast. These findings highlight the ability of C. glabrata to manipulate host proteins involved in endocytic processes and intracellular trafficking. Likewise, these results suggest a strong role of host autophagy in controlling fungal pathogens such as C. glabrata.

Candida parapsilosis is an opportunistic pathogen with increasing rates of resistance to fluconazole and voriconazole. Recently, in an outbreak at the Azienda Ospedaliero-Universitaria Pisana, a new amino acid substitution, S862C in the CpMrr1 protein, was found only in azole-resistant strains. The contribution of this mutation to the acquisition of an azole-resistant phenotype was investigated in this study.

Antifungal resistance in C. parapsilosis clinical strains isolated from the outbreak (n = 16) was tested by the broth microdilution method and Etest strip. WGS and Sanger sequencing analyses were used for the detection of SNPs. A CRISPR-Cas9-based genome editing strategy was used to induce the C2585G substitution in the CpMRR1 gene of susceptible C. parapsilosis isolates to investigate its role in the acquisition of azole resistance.

The A395T and the newly found C2585G substitution in the CpMRR1 gene were present in all resistant isolates, but not in the susceptible ones. Such mutations were later induced in the C. parapsilosis reference strain ATCC 22019 and in two azole-susceptible clinical isolates in homozygosis, and in heterozygosis only for ATCC 22019 and one azole-susceptible clinical isolate. Both heterozygous and homozygous mutants carrying the C2585G mutation were fluconazole resistant, with some clones also presenting intermediate susceptibility or resistance to voriconazole.

To the best of our knowledge, this is the first study to report the effect on azole resistance of a novel C2585G nucleotide substitution in the CpMRR1 gene found in clinical isolates recovered during an outbreak of azole-resistant C. parapsilosis in a healthcare setting.

To investigate the epidemiology and molecular typing of 307 clinical Wickerhamomyces anomalus isolates collected in China.

A total of 307 W. anomalus isolates were collected from CHIF-NET, a surveillance network with nationwide coverage, from 2009 to 2021. Antifungal susceptibility of all W. anomalus isolates were tested by broth microdilution according to CLSI methods. Genotyping of all isolates was performed using a panel of polymorphic microsatellite markers.

The number of W. anomalus isolates was highest in Northeast China, with 121 strains, accounting for 4.6% of the total Candida spp. isolates in the region. Newborns emerged as the primary community of infection, with 84 isolates (12.7% of all Candida spp. in the newborn group). Most W. anomalus isolates were recovered from blood samples (249 isolates). The isolates were found to exhibit notable ratios of non-wild type to fluconazole and voriconazole (48.5% and 34.5%). A total of 118 microsatellite-based types were identified among a set of 309 isolates that included the 307 clinical isolates along with reference strains ATCC 8168 and CICC 32553. This analysis revealed potential nosocomial outbreaks.

Our research is the largest investigation of W. anomalus drug susceptibility thus far, providing foundational data that could contribute to future establishment of resistance breakpoints and enhancing epidemiological surveillance capabilities. Our study suggests the importance of nosocomial surveillance and drug resistance management for W. anomalus.

Invasive candidiasis has emerged as a significant healthcare challenge, with a rising incidence rate attributed to the widespread use of organ transplantation, chemotherapy, immunosuppressants, and broad-spectrum antibiotics. The increasing prevalence of drug-resistant strains, particularly among Candida tropicalis, has necessitated the exploration of novel therapeutic strategies. Our study investigated the synergistic effects of minocycline (MIN) combined with fluconazole (FLC) against FLC-resistant C. tropicalis, both in vitro and in vivo. The in vitro synergistic activity of MIN and FLC was evaluated using checkerboard titration and time-kill assays. The Galleria mellonella larvae and mouse model were employed to assess in vivo efficacy, with histopathological examination and fungal burden quantification. Whole-genome and RNA sequencing elucidated the synergistic mechanisms observed. The FLC/MIN combination significantly lowered the minimum inhibitory concentration (MIC) and improved fungicidal activity, as evidenced by enhanced survival rates and reduced fungal burden in G. mellonella larvae and mouse models. Histopathological analysis confirmed less tissue damage and fungal load with combination therapy. RNA sequencing analysis suggested that the impact of MIN on amino acid metabolism contributes to the synergistic effects. This approach holds promise for treating FLC-resistant C. tropicalis by increasing antifungal efficacy and reducing drug resistance risks, warranting further clinical exploration.

This study highlights the potential of minocycline and fluconazole combination therapy in combating drug-resistant Candida tropicalis. It shows promising in vitro and in vivo synergistic effects, reducing MIC and enhancing fungicidal activity. Further clinical trials are needed to validate its efficacy in treating FLC-resistant C. tropicalis infections.

The recent rise in the global incidence of fluconazole resistance in C. parapsilosis has become a significant public health concern. Epidemiological studies suggest that fluconazole resistance in C. parapsilosis spreads through endemic clones. We, therefore, investigated the molecular epidemiology of fluconazole-resistant C. parapsilosis in our centre.

C. parapsilosis isolates from 2016 through 2022 were investigated for antifungal susceptibility. Fluconazole-resistant isolates were analysed for ERG11 mutation using Sanger sequencing. Gene expression profiles of ERG11, CDR1 and MDR1 were assessed by real-time qPCR. The epidemiological relationship of resistant and susceptible isolates of C. parapsilosis was investigated using short tandem repeat typing. Additionally, biofilm production and cell wall ergosterol contents were also quantified and compared.

Among 572 C. parapsilosis isolates, 48 (8.4%) were resistant to fluconazole. Of 28 recoverable resistant isolates, 17.9% (5/28) were wild-type and 82.1% (23/28) harboured the following ERG11 mutations: Y132F (n = 3), K143R (n = 10) and K143R + R398I (10/28). Significant fold-changes were observed in ERG11 (p = 0.037) and MDR1 (p = 0.008) gene expressions in fluconazole resistant compared to susceptible isolates. Contrary to global reports, STR typing suggested a limited clonal transmission of resistant C. parapsilosis with multiple introductions of resistant isolates in our centre. On fluconazole exposure, ergosterol content significantly increased (p < 0.01) in resistant isolates, particularly in isolates harbouring ERG11K143R + R398I mutations. In contrast, fluconazole-susceptible isolates formed comparatively higher baseline biofilm (p < 0.05) than resistant isolates with ERG11K143R mutation.

The current study underscores the need for continuous molecular surveillance and tailored therapeutic options for effective management of fluconazole resistance in C. parapsilosis.

The emergence of COVID-19 has led to a significant public health crisis, and an increase in fungal infections, including candidemia. Candida species are frequently found in intensive care units (ICUs), and it is a common cause of death in many patients. The isolates were identified using polymerase chain reaction-restriction. In this study, We investigated the factors linked to Candida infections in COVID-19 burn patients in the ICU and assessed the antifungal susceptibility of the isolates in vitro.

Out of 335 burn patients admitted to the ICU, fifty-six with concurrent COVID-19 were included in this study. A total of 133 yeast isolates were obtained from burn wounds, 29 from blood cultures, and 36 from urine cultures. The isolates were identified using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis.

Out of fifty-six patients, twenty-nine had infections and forty-eight had colonization, with Candida parapsilosis being the most common species. Twenty-one patients died during their ICU stay, with mortality rates of 43.8% among colonized patients and 69.0% among infected patients. Fluconazole and itraconazole exhibited the highest minimum inhibitory concentrations, while luliconazole and amphotericin B were identified as the most effective antifungal agents.

Our findings indicate that colonization may act as an important prognostic factor prior to the onset of candidemia. In addition, prolonged hospitalization, catheter use, and concurrent COVID-19 infection were identified as key risk factors for candidemia in this patient group. Notably, the rising drug resistance in non-albicans Candida species is a major public health concern.

The dynamic chromatin structure regulates many biological processes including gene expression, DNA repair, and genome stability in eukaryotic cells. However, its role in governing antifungal drug susceptibility in medically important fungi is just beginning to be deciphered. Chromatin architecture is maintained by a complex interplay among histone protein stoichiometry sustainment, post-translational modifications of histone proteins, and the activity of chromatin remodeling complexes. Herein, we report that the reduced gene dosage of histone core proteins in the opportunistic human fungal pathogen Candida glabrata leads to increased susceptibility toward the widely used, cell wall-targeting echinocandin antifungal drugs. Our comprehensive characterization of single and double histone mutants revealed that linker histone H1 loss had no effect on cell physiology and drug susceptibility, whereas low H2A, H2B, H3, and H4 protein levels resulted in decreased reactive oxygen species production, altered biofilm production, elevated DNA damage, and echinocandin stress susceptibility. Importantly, not all core histone mutants exhibited an increased sensitivity to other cell wall stressors, thereby precluding a general cell wall defect accounting solely for the increased caspofungin susceptibility. Finally, we show that the histone H3 acetylation at lysine-56 may be pivotal to caspofungin response of C. glabrata, as H3K56Ac levels were reduced in both core histone mutants and upon caspofungin exposure, with H3K56 acetyltransferase (CgRtt109)- and nucleosome assembly factor (CgAsf1)-lacking mutants displaying increased caspofungin susceptibility. Besides demonstrating the histone requirement for the survival of C. glabrata in the mouse systemic candidiasis model, our findings unveil histone dosage-regulated cellular processes that impact echinocandin susceptibility.

Echinocandin antifungals, which impede cell wall synthesis, are often used to treat Candida bloodstream infections. The human opportunistic fungal pathogen Candida (Nakaseomyces) glabrata is increasingly being reported to exhibit co-resistance to echinocandins and ergosterol biosynthesis-inhibitory azole drugs in hospitals worldwide. However, the role of histones, protein-building blocks of the nucleosome, in governing echinocandin resistance in C. glabrata is not understood. Herein, we show that the reduced gene dosage of core histone proteins, but not of the linker histone, leads to echinocandin susceptibility, which is partly due to increased ROS levels. Additionally, our data implicate histone H3 acetylation at lysine-56 in the caspofungin response of C. glabrata. Since the emerging echinocandin resistance is an impediment to successful antifungal therapy, our findings open up a new research avenue of pharmacological targeting of histone proteins that could potentially block echinocandin resistance and attenuate C. glabrata survival in the host.

The morbidity and mortality of invasive fungal infections are increasing rapidly. Developing effective and safe antifungal drugs with novel chemical scaffolds and mechanisms is urgently needed. On the basis of our previously identified Pdr1-KIX inhibitor 1, a series of new pyrazolone-carbothioamide derivatives were designed and assayed. In particular, compound A7 showed picomolar in vitro antifungal activity against Candida glabrata (MIC = 0.00012 μg/mL) and Cryptococcus neoformans (MIC = 0.00012 μg/mL), with excellent antivirulence effects. In the murine candidiasis and cryptococcosis models, compound A7 exhibited potent in vivo therapeutic efficacy. Interestingly, a mechanism investigation revealed that the antifungal activity of compound A7 is independent of KIX binding. It disrupted the iron homeostasis of fungal cells and then induced oxidative stress damages by accumulating the reactive oxygen species and lipid peroxides. Therefore, compound A7 represents a promising lead with a new mechanism of action to combat candidiasis and cryptococcosis.

Vertebrate lungs contain diverse microbial communities, but little is known about the drivers of community composition or consequences for health. Microbiome assembly by processes such as dispersal, coevolution, and host-switching can be probed with comparative surveys; however, few studies exist for lung microbiomes, particularly for the fungal component, the mycobiome. Distinguishing among fungal taxa that are generalist or specialist symbionts, potential pathogens, or incidentally inhaled spores is urgent because of potential for emerging diseases. Here, we characterize the avian lung mycobiome and test the relative influences of environment, phylogeny, and functional traits. We used metabarcoding and culturing from 195 lung samples representing 32 bird species across 20 families. We identified 526 fungal taxa as estimated by distinct sequence types (zOTUs) including many opportunistic pathogens. These were predominantly from the phylum Ascomycota (79%) followed by Basidiomycota (16%) and Mucoromycota (5%). Yeast and yeast-like taxa (Malassezia, Filobasidium, Saccharomyces, Meyerozyma, and Aureobasidium) and filamentous fungi (Cladosporium, Alternaria, Neurospora, Fusarium, and Aspergillus) were abundant. Lung mycobiomes were strongly shaped by environmental exposure, and further modulated by host identity, traits, and phylogenetic affinities. Our results implicate migratory bird species as potential vectors for long-distance dispersal of opportunistically pathogenic fungi.

Fungi are important and often underestimated human pathogens. Infections with fungi mostly originate from the environment, from soil or airborne spores. By contrast, Candida albicans, one of the most common and clinically important fungal pathogens, permanently exists in the vast majority of healthy individuals as a member of the human mucosal microbiota. Only under certain circumstances will these commensals cause infections. However, although the pathogenic behaviour and disease manifestation of C. albicans have been at the centre of research for many years, its asymptomatic colonization of mucosal surfaces remains surprisingly understudied. In this Review, we discuss the interplay of the fungus, the host and the microbiome on the dualism of commensal and pathogenic life of C. albicans, and how commensal growth is controlled and permitted. We explore hypotheses that could explain how the mucosal environment shapes C. albicans adaptations to its commensal lifestyle, while still maintaining or even increasing its pathogenic potential.

The emergence of azole-resistant Candida infections is a major concern. A key mechanism is the gain of resistance through amino acid substitutions in the sterol 14α-demethylase, the main target of azole drugs. While numerous resistant substitutions are known, the pattern of such substitutions remains unclear. We hypothesized that resistant substitutions occur disproportionately at azole-binding sites. We compiled 2222 instances of azole-resistant substitutions from the literature and performed extensive computational sequence analyses. Altogether, there were 169 known substitutions at 133 sites in sterol 14α-demethylases of seven Candida species, whereas C. albicans alone had 120 substitutions at 97 sites. Just 10 sites and 18 substitutions (such as Y132F/H, K143R, D116E, and G464S) accounted for 75% of the total instances. Only about 48% of the sites were present within previously recognized hotspot regions, while just 33% of the azole-interacting residues had known resistant substitutions, most of them with only a few instances. The literature data on azole-resistant substitutions in Candida appear to be highly biased, as a few substitutions, such as Y132F/H and K143R, were preferentially sought and reported with over 1,000 instances. Additionally, there were numerous reports of "resistant" substitutions in azole-susceptible Candida isolates. Our study provides new perspectives into azole resistance.

Candida glabrata (Nakaseomyces glabratus) is the most common cause of drug-resistant candidemia and is associated with a high mortality rate. Only a few mechanisms of drug resistance are known in C. glabrata, predominantly involving recurrent single nucleotide polymorphisms. The importance of structural variation in acquired drug resistance is not understood. We performed comparative phenotypic and genomic analyses of six serial bloodstream isolates of C. glabrata and identified novel mutations associated with resistance to echinocandins. Critically, we identified a novel gene conversion event between the hotspot 2 regions of FKS1 and FKS2 that was associated with increased resistance to micafungin. We further analyzed 621 publicly available C. glabrata genomes and found three additional examples of structural variation involving FKS1 and FKS2. Ultimately, drug resistance in C. glabrata involves structural variants that are missed with current diagnostic methods and need to be considered when designing and implementing more effective antifungal management strategies.

Invasive fungal infections are responsible for millions of deaths worldwide each year. Therefore, focusing on innovative approaches to developing therapeutics that target fungal pathogens is critical. Here, we discuss targeting the fungal trehalose biosynthesis pathway with antifungal therapeutics, which may lead to the improvement of human health globally, especially as fungal pathogens continue to emerge due to fluctuations in the climate.

Candida albicans is a major cause of systemic candidiasis, a severe fungal infection with a ∼40% mortality rate. Yck2, a casein kinase 1 (CK1) in C. albicans, is targeted by antifungal inhibitors YK-I-02 (YK) and MN-I-157 (MN). Using multiplexed inhibitor beads and mass spectrometry (MIB/MS), the selectivity of these inhibitors was determined across the fungal kinome. The MIB matrix captured 89% of C. albicans protein kinases, revealing that YK and MN selectively engage three CK1 homologues (Yck2, Yck22, and Hrr25) and a human p38α homologue (Hog1). Chemoproteomics using a custom MN-kinobead confirmed the remarkable fungal kinome selectivity. To identify new Yck2 inhibitors with selectivity over Hog1, 13 human CK1 inhibitors were screened, leading to the discovery of a new chemotype with antifungal activity. These findings highlight the utility of MIB/MS in profiling nonhuman kinomes and developing selective fungal kinase inhibitors as antimicrobial agents.

In recent years, there has been an increase in the incidence of fluconazole-non-susceptible (FNS) Candida parapsilosis. The reasons why these strains are able to colonize hospitals remain unknown. It is also unclear whether these strains exhibit resistance to the disinfectants used in hospitals, facilitating their spread. For these reasons, in this work, we aimed to investigate whether fluconazole resistance was associated with virulence traits and the resistance of these strains to common hospital disinfectants. The general conclusion of the study was that more than 95% of the FNS strains, regardless of the resistance mutation they carried, had filamentation problems, whereas around 75% of the susceptible strains formed pseudohyphae and were capable of filamentation. This 95% of the FNS strains did not form pseudohyphae, did not invade agar, and did not form biofilms, while the susceptible strains exhibited the opposite behaviour. Through microfluidics experiments, we observed that both the susceptible and FNS strains were capable of adhering to a plastic surface under dynamic conditions, but the FNS strains formed unstable aggregates that did not remain attached to the surface, confirming the filamentation defect of these strains. In the second part of the study, we observed that FNS strains are susceptible to clinical disinfectants, although they presented a slight resistance to some of them, such as chlorhexidine, compared to susceptible isolates. In this work, we address important aspects to understand the dissemination of FNS strains in clinical outbreaks.

Candida glabrata is the second most common species responsible for invasive candidiasis, including candidaemia. Echinocandins are typically the first-line therapy for C. glabrata candidaemia, with the option to transition to oral fluconazole. Studies are needed to evaluate clinical outcomes in patients initially treated with echinocandins then transitioned to fluconazole.

This was a retrospective, single-centre cohort study of patients with C. glabrata candidaemia from November 2011 to August 2023. Inpatients aged 18-89 years who received an echinocandin within 24 h of the initial positive blood culture were included. Patients were excluded if they received antifungal treatment less than 48 h, combination therapy, or fluconazole as initial therapy. The primary composite outcome was 30-day clinical failure.

A total of 186 patients were included (n = 153 echinocandin only; n = 33 fluconazole step-down). The most common source of candidaemia was line-associated in both groups with the majority having source control (43% echinocandin versus 58% fluconazole; P = 0.32). Compared to fluconazole, patients in the echinocandin group had a higher rate of concomitant bacteraemia (45% versus 24%; P = 0.03) and endovascular complications (11% versus 0%; P = 0.05). There was no significant difference in treatment duration between echinocandin and fluconazole (16 versus 19 days; P = 0.46), incidence of persistent candidaemia (22% versus 24%; P = 0.7), or 30-day clinical failure (15% versus 9%; P = 0.58).

Fluconazole appears to be a safe and reasonable step-down therapy in the management of C. glabrata candidaemia.

Cyberlindnera fabianii, an ascomycetous yeast, is a rare human pathogen. Here, we describe the first reported case of central venous (CV) access port-related blood-stream infection caused by C. fabianii. An 80-year-old Asian man presented to our department with a mass in the pancreas. A diagnosis of pancreatic cancer (cT4N0M0, cStage III) was made. A catheter with CV access port was inserted, and anticancer chemotherapy and radiotherapy was initiated. One year after the first visit, he was urgently admitted to our hospital due to recurrent hematemesis. On day 37, organism cultures of the blood and catheter tip were positive for Candida pelliculosa by the BD PHOENIX™ 100 system. On the basis of these findings, CV port-related blood-stream infection caused by C. pelliculosa was diagnosed, and treatment with micafungin (MCFG) was initiated. On day 51, the blood culture converted to negative. On day 65, MCFG was stopped. The cultured strain was sent to reference laboratory where Cyberlindnera fabianii was identified by MALDI-TOF mass spectrometry and direct sequencing of the internally transcribed spacer region. The present case report describes the first known case of CV access port-related blood-stream infection caused by C. fabianii. MCFG was successfully used to treat the fungemia caused by C. fabianii. Although the incidence of uncommon yeast species causing human infections has gradually increased in recent years, conventional biochemical methods may offer limited usefulness for identifying these rare organisms. Accordingly, MALDI-TOF mass spectrometry or molecular assays may be required to identify these uncommon fungal species.

Candida glabrata is a common causative pathogen of endogenous candidiasis. It is assumed that the gastrointestinal flora affects C. glabrata gastrointestinal colonization and organ dissemination in the gastrointestinal tract (GIT). However, no reports have yet described the relationships between C. glabrata and bacteria in the GIT. This study aimed to clarify these relationships using a mouse endogenous candidiasis model with cortisone acetate immunosuppression.

Dysbiosis was induced in the GIT by several antibiotic combinations, and then C. glabrata gastrointestinal colonization and organ dissemination were evaluated. Next, metagenomic sequencing analysis of the gastrointestinal flora was performed to identify bacteria associated with C. glabrata organ dissemination. Finally, coinfection experiments were performed using bacteria isolated from the mouse GIT.

C. glabrata organ dissemination was significantly promoted using specific antibiotics regardless of the amount of colonization in the GIT. Metagenomic sequencing analysis of the gastrointestinal flora showed that Enterococcus species and anaerobes were significantly associated with enhanced organ dissemination, whereas Enterobacterales, such as Escherichia species and Klebsiella species, were associated with the suppression of organ dissemination. In coinfection experiments, Enterococcus faecalis and Faecalibaculum rodentium inoculation, but not either of them, increased C. glabrata organ dissemination without affecting gastrointestinal colonization.

Coinfection with gastrointestinal bacteria promoted C. glabrata organ dissemination, which would indicate that gastrointestinal flora could affect C. glabrata dissemination. Therefore, the gastrointestinal flora could be a target for intervention or treatment in clinical settings. Insights from this study would lead to better control of endogenous candidiasis focusing on the gastrointestinal flora.

Invasive fungal infections, significantly impact hospitalized and immunocompromised populations. Recent trends showed a shift from Candida albicans to non-albicans Candida (NAC) species, raising concerns about antifungal resistance.

Our study focuses on the distribution of fungal species in blood cultures obtained from different healthcare settings, including hospitals, long-term care facilities, and community health centers in the Venetian region of Italy.

We retrospectively analyzed all consecutive blood culture isolates across 5 hospitals, 38 long-term care facilities, and 24 sample collection centers (blood exams and culture) from 2019 to 2023.

Between 2019 and 2023, 11,552 microorganisms were isolated from blood cultures; 693 (6.0%) were fungi. The yearly prevalence ranged from 5.2% in 2019 to 6.1% in 2023. C. albicans isolates decreased significantly, from 60.0% in 2019 to 43.1% in 2023. NAC species showed significant growth, particularly C. parapsilosis sensu stricto (from 23.6% in 2019 to 28.8% in 2023), C. tropicalis (from 0.0% in 2019 to 7.2% in 2023), and N. glabratus (from 9.1% in 2019 to 11.8% in 2023). Medical wards consistently recorded the highest number of cases (429/693, 61.9%), with C. albicans predominating in earlier years. Resistance to amphotericin B rose sharply in C. parapsilosis ss. (22.5% in 2022), while fluconazole resistance in N. glabratus remained high (peaking at 85.7% in 2021).

The increasing dominance of NAC species and rising resistance trends underscore the necessity for enhanced diagnostics, infection prevention, and antifungal stewardship. Future research should incorporate clinical data to optimize fungemia management strategies.

Bloodstream infections (BSIs) are common in hospitals, often life-threatening and increasing in prevalence. Microorganisms in the blood are usually rapidly cleared by the immune system and filtering organs but, in some cases, they can cause an acute infection and trigger sepsis, a systemic response to infection that leads to circulatory collapse, multiorgan dysfunction and death. Most BSIs are caused by bacteria, although fungi also contribute to a substantial portion of cases. Escherichia coli, Staphylococcus aureus, coagulase-negative Staphylococcus, Klebsiella pneumoniae and Candida albicans are leading causes of BSIs, although their prevalence depends on patient demographics and geographical region. Each species is equipped with unique factors that aid in the colonization of initial sites and dissemination and survival in the blood, and these factors represent potential opportunities for interventions. As many pathogens become increasingly resistant to antimicrobials, new approaches to diagnose and treat BSIs at all stages of infection are urgently needed. In this Review, we explore the prevalence of major BSI pathogens, prominent mechanisms of BSI pathogenesis, opportunities for prevention and diagnosis, and treatment options.

Candida auris has emerged as a multidrug-resistant yeast implicated in healthcare-associated invasive infections and hospital outbreaks. The aim of the current 38-month period observational study in a multidisciplinary Intensive Care Unit (ICU) was to analyze the epidemiology, potential risk factors, management strategies, and patient outcomes of patients with C. auris. During the study period, 32 patients were identified with C. auris infection (6 patients) or colonization (26 patients) and their clinical characteristics and treatment-related factors were compared. Identification of C. auris isolates was confirmed by MALDI-TOF spectrometry. According to our results, regarding patient-related factors, no significant differences were identified. Regarding treatment-related factors, the proportion of patients already receiving corticosteroids (34.6% vs. 83.3%, p = 0.064) or being on renal replacement treatment (7.7% vs. 33.3%) was higher in infected patients. Median time elapsed from ICU admission to first positive culture was 7 (1-21) days and half of cases were ICU-imported. All strains were resistant to fluconazole and susceptible to echinocandines and amphotericin B. Crude mortality of the study population was 43.75%, similar to other previously reported candidemias. Rapid identification of C. auris, continued surveillance, and infection control practices are important elements for controlling successfully its spread in the hospital setting and for establishing promptly its transition from commensalism to infection.

Candida spp. infections have increasingly been reported among COVID-19 patients, yet the epidemiological factors, diagnostic methods, and outcomes associated with these infections remain poorly understood. These infections, particularly in ICU settings, present significant challenges due to high mortality rates and rising antifungal resistance. This study aimed to investigate the occurrence, risk factors, treatment, and outcomes of Candida albicans and non-albicans Candida in COVID-19 patients, providing clinical and epidemiological insights.

A review following PRISMA guidelines was conducted. Searches were performed in PubMed, Embase, and BVS databases, covering articles published from January 2020 to May 2024. Inclusion criteria included case reports or case series providing detailed information on Candida spp. in COVID-19 patients. Data extraction focused on patient demographics, underlying diseases, antifungal and antibiotic therapies, antifungal susceptibility, resistance profiles, and outcomes. Statistical analyses were conducted using SPSS software.

The review included 67 studies, totaling 223 COVID-19 patients. Male patients were predominant. Common comorbidities included hypertension, cancer, and dyslipidemia. Echinocandins were the primary antifungal treatment. Non-albicans Candida exhibited a higher resistance rate (47.10 %) compared to C. albicans (2.35 %). Overall mortality rates were high, at 60.50 % for C. albicans and 62.30 % for non-albicans. Significant risk factors for mortality included age, central venous catheter use, ICU admission, and corticosteroid therapy.

The study identified critical risk factors and clinical characteristics in COVID-19 patients with Candida infections. The high incidence of antifungal resistance among non-albicans and high mortality rates highlight the need for vigilant monitoring and targeted antifungal strategies to improve outcomes.

The Candida Genome Database (CGD; www.candidagenome.org) is unique in being both a model organism database and a fungal pathogen database. As a fungal pathogen database, CGD hosts locus pages for 5 species of the best-studied pathogenic fungi in the Candida group. As a model organism database, the species Candida albicans serves as a model both for other Candida spp. and for non-Candida fungi that form biofilms and undergo routine morphogenic switching from the planktonic form to the filamentous form, which is not done by other model yeasts. As pathogenic Candida species have become increasingly drug resistant, the high lethality of invasive candidiasis in immunocompromised people is increasingly alarming. There is a pressing need for additional research into basic Candida biology, epidemiology and phylogeny, and potential new antifungals. CGD serves the needs of this diverse research community by curating the entire gene-based Candida experimental literature as it is published, extracting, organizing, and standardizing gene annotations. Gene pages were added for the species Candida auris, recent outbreaks of which have been labeled an "urgent" threat. Most recently, we have begun linking clinical data on disease to relevant Literature Topics to improve searchability for clinical researchers. Because CGD curates for multiple species and most research focuses on aspects related to pathogenicity, we focus our curation efforts on assigning Literature Topic tags, collecting detailed mutant phenotype data, and assigning controlled Gene Ontology terms with accompanying evidence codes. Our Summary pages for each feature include the primary name and all aliases for that locus, a description of the gene and/or gene product, detailed ortholog information with links, a JBrowse window with a visual view of the gene on its chromosome, summarized phenotype, Gene Ontology, and sequence information, references cited on the summary page itself, and any locus notes. The database serves as a community hub, where we link to various types of reference material of relevance to Candida researchers, including colleague information, news, and notice of upcoming meetings. We routinely survey the community to learn how the field is evolving and how needs may have changed. For example, we asked our users which species we should next add to CGD, and the clear answer was Candida tropicalis. A key future challenge is management of the flood of high-throughput expression data to make it as useful as possible to as many researchers as possible. The central challenge for any community database is to turn data into knowledge, which the community can access, use, and build upon.

This study assesses the impact of fluconazole resistance on 30-day all-cause mortality and 1-year recurrence in patients with Candida parapsilosis bloodstream infections (BSI).

A multicenter retrospective study was performed at 3 hospitals in Italy and Spain between 2018 and 2022. Adult patients with positive blood cultures for C. parapsilosis who received appropriate targeted therapy with either echinocandins or fluconazole were included.

Among 457 patients, 196 (42.9%) had fluconazole-resistant C. parapsilosis (FLZR-CP) BSI and 261 (57.1%) had fluconazole-susceptible C. parapsilosis (FLZS-CP) BSI. All FLZR-CP patients received targeted echinocandins, while FLZS-CP patients received either echinocandins (60.5%) or fluconazole (39.5%). Unadjusted 30-day all-cause mortality rates were 28.6% for FLZR-CP and 28.4% for FLZS-CP (log-rank test, P = .998). In multivariable analysis, increased mortality was associated with age (adjusted hazard ratio [aHR] 1.03 per year; 95% confidence interval [CI], 1.01-1.05; P = .0005), solid tumor (aHR 1.91; 95% CI, 1.06-3.46; P = .0302), previous antifungal treatment (aHR 1.84; 95% CI, 1.12-3.10; P = .0192), and septic shock (aHR 2.39; 95% CI, 1.42-4.06; P = .0010), but not fluconazole resistance (aHR 1.00; 95% CI, .62-1.63; P = .9864) nor the type of initial antifungal therapy (aHR 1.46; 95% CI, .69-3.06; P = .3202). Propensity score-matched analysis showed no 30-day all-cause mortality difference between echinocandin-treated FLZR-CP and fluconazole-treated FLZS-CP patients (HR 0.81; 95% CI, .37-1.75; P = .5915). However, a higher 1-year recurrence risk was observed in FLZR-CP patients (odds ratio, 7.37; 95% CI, 2.11-25.80; P = .0018).

Our results suggest that fluconazole resistance is not associated with a higher mortality risk in patients with C. parapsilosis BSI, though 1-year recurrence rates were higher in the FLZR-CP group.

Candida species are major pathogens of bloodstream infections (BSIs) in hospitalized patients, with high mortality. This study examined Candida species distribution, clinical characteristics, and the mortality of patients with Candida BSIs. Adult patients (≥16 years) with Candida BSIs at a teaching hospital (2014-2023) were retrospectively reviewed. Over 10 years, 487 Candida isolates were obtained from 462 patients. C. albicans was the most frequent (38.2%), followed by C. glabrata (21.1%), C. parapsilosis (20.5%), and C. tropicalis (13.3%). The annual incidence of Candida BSIs remained stable (p = 0.525). However, non-albicans species BSIs increased 1.61-fold compared to C. albicans (95% CI: 1.19-2.19, p = 0.002). Fluconazole-non-susceptible Candida isolates increased after 2021 (p = 0.040). The overall 30-day mortality was 40.6%. In the multivariate analysis, a high Charlson comorbidity index (aHR: 1.20, 95% CI: 1.07-1.35, p = 0.001) and high SOFA score (aHR: 1.12, 95% CI: 1.02-1.23, p = 0.022) were the strongest predictors of 30-day mortality. Meanwhile, C. parapsilosis BSIs (aHR: 0.46, 95% CI: 0.22-0.99, p = 0.047) and central venous catheter removal at any time (aHR: 0.22, 95% CI: 0.13-0.37, p < 0.001) were associated with reduced 30-day mortality. The mortality of patients with Candida BSIs was mainly determined by disease severity, while catheter removal was associated with improved survival.

Pichia kudriavzevii causes life-threatening infections in immunocompromised hosts, including hospitalized neonates. This pathogen is intrinsically resistant to fluconazole, while uncommon P. kudriavzevii strains resistant to multiple antifungal drugs, including voriconazole, amphotericin B, and echinocandins, have also been reported from healthcare environments. Thus, understanding how P. kudriavzevii spread, persist, and adapt to healthcare settings could help us develop better infection management strategies. In this study, whole genome sequencing identifies multiple outbreaks of bloodstream infections in a single neonatal intensive care unit (NICU) over 5 years caused by genetically diverse strains of P. kudriavzevii. Interestingly, two genetically distinct clusters of P. kudriavzevii strains showed frequent loss of heterozygosity (LOH) events between two temporal samples. The first outbreak cluster (2015-2016) showed LOH at chromosomes 1, 4, and 5, and the other outbreak cluster (2020) exhibited LOH at chromosome 2. The circulation of two separate strain clusters of P. kudriavzevii suggests nosocomial transmission in the NICU in different time periods. Furthermore, we compared the transcriptomic profiles of three isolates of clusters I and II that exhibited distinct fluconazole and itraconazole MICs. While no significant difference in gene expression was found at the azole-target gene ERG11 or the ATP-binding cassette (ABC) transporter genes, such differences were found in genes involved in cell division and filamentation, such as SIR2 (sirtuin deacetylase) and RFA1 (replication factor A). Interestingly, increased filamentation was observed in clade I isolate exhibiting high fluconazole MICs. Together, our study indicates significant diversity, persistence, and rapid evolution of P. kudriavzevii within a single NICU.

The rising burden of fungal infections presents a significant challenge to global healthcare, particularly with increasing antifungal resistance limiting treatment efficacy. Early detection and timely intervention remain critical, yet fungal pathogens employ diverse mechanisms to evade host immunity and develop resistance, undermining existing therapeutic options. Limited antifungal options and rising resistance necessitate novel treatment strategies. This review provides a comprehensive overview of conventional antifungal agents, their mechanisms of action, and emerging resistance pathways. Furthermore, it highlights recently approved and investigational antifungal compounds while evaluating innovative approaches such as nanotechnology, drug repurposing, and immunotherapy. Addressing antifungal resistance requires a multifaceted strategy that integrates novel therapeutics, enhanced diagnostic tools, and future research efforts to develop sustainable and effective treatment solutions.

Candida albicans is the most common cause of life-threatening fungal infection in the developed world but remains a therapeutic challenge. Protein kinases have been rewarding drug targets across diverse indications but remain untapped for antifungal development. Previously, screening kinase inhibitors against C. albicans revealed a 2,3-aryl-pyrazolopyridine, GW461484A (GW), which targets casein kinase 1 (CK1) family member Yck2. Here, we report optimization of GW via two complementary approaches, synthesis of bioisosteres possessing an imidazo[1,2-a]pyridine core, and R-group substitution of GW's pyrazolo[1,5-a]pyridine core. Characterization of compounds reveals two 6-cyano derivatives with improved pharmacological properties that retain whole-cell bioactivity and selectivity for fungal Yck2 compared to human CK1α. Efficacy studies in mice indicate both analogs possess single-agent activity against C. albicans resistant to first-line echinocandin antifungals and potentiate non-curative echinocandin treatment. Results validate Yck2 as an antifungal target and encourage further development of inhibitors acting by this previously unexploited mode of action.

Prevention and management of invasive fungal infections is challenging due to the complexity of at-risk patient population, high morbidity and mortality of these infections, and pharmacologic aspects of available antifungal agents. While there has been substantial investment in antifungal drug development over past 20 years, the ideal antifungal remains elusive. Clinicians must be aware of differences in spectrum of activity, pharmacokinetic/dynamic dosing, toxicity, resistance, and drug interaction profiles of antifungals to use them most effectively. This article will review key features of U.S. Food and Drug Administration-approved and pipeline antifungals to facilitate an understanding of their role in treatment and/or prevention of invasive fungal infections.

Invasive candidiasis (IC) is a term that refers to a group of infectious syndromes caused by a variety of Candida species, 6 of which cause the vast majority of cases globally. Candidemia is probably the most commonly recognized syndrome associated with IC; however, Candida species can cause invasive infection of any organ, especially visceral organs, vasculature, bones and joints, eyes, and central nervous system. The optimal use of these newer diagnostics coupled with a thoughtful clinical assessment of at-risk patients and the judicious use of effective antifungal therapy is a key to achieving good antifungal stewardship and improved patient outcomes.

Among the Candida species commonly involved in superficial and more significant life-threatening infections, C. krusei exhibits the most worrisome resistance profile to antifungals. This study aimed to analyse the population structure using multilocus sequence typing (MLST), and to evaluate the antifungal susceptibility profile of C. krusei isolated from patients living with human immunodeficiency virus (HIV) in Cameroon. C. krusei isolated from stool, urine, mouth and vaginal samples were identified using routine laboratory techniques and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). The C. krusei isolates were further analysed by MLST. In vitro antifungal susceptibility testing was performed using the Sensititre Yeast One™ microdilution technique. Forty three (43) C. krusei isolates were included in the study. The MLST identified 32 Diploid sequence type (DST), of which 31 were new that were not included in the current database. New alleles were not observed. Different DSTs were observed in isolates from the same geographical area, from different anatomical sites in the same patient. An eBURST analysis clustered all identified DSTs of former isolates in clonal complex 1. Heterogeneous antifungal MICs were observed in isolates of the same DST and/or from the same geographical area. 32.6% of the isolates displayed a resistant or non-wild-type phenotype to at least 3 distinct antifungal agent classes. The achieved results support the setting up of molecular epidemiology and antifungal resistance surveillance of C. krusei.

This study investigated the distribution and antibiotic resistance profiles of common bacteria isolated from clinical specimens at a hospital's microbiology laboratory between 2020 and 2022.

A retrospective analysis was conducted on microbial culture results from clinical specimens collected over three years, including sample types, departmental distribution, pathogen species, and resistance profiles.

A total of 13,048 unique pathogenic strains were isolated, predominantly from respiratory and urine specimens. Secretion specimens exhibited the highest positive detection rate (73.6%), while blood specimens showed a lower rate (9.7%). The five most frequently isolated pathogens were: Klebsiella pneumoniae (K. pneumoniae) (19.6%), Pseudomonas aeruginosa (P. aeruginosa) (14.7%), Escherichia coli (E. coli) (9.2%), Acinetobacter baumannii (A. baumannii) (8.0%), and Candida albicans (C. albicans) (7.0%). Gram-negative bacteria constituted 53.7% of all isolates (7009/13,048). A total of 7590 multidrug-resistant organisms (MDRO) were identified, corresponding to a detection rate of 21.3% (7590/35,613). The detection rates of carbapenem-resistant Enterobacteriaceae (CRE) increased annually: 7.2% (2020), 8.6% (2021), and 14.4% (2022).

The annual detection rate of CRE increased during the study period, while the rate of methicillin-resistant Staphylococcus aureus (MRSA) declined. Timely and effective interventions targeting pathogenic bacteria are essential for controlling and mitigating nosocomial infection risks.

Candida infections severely impact patients who are immunocompromised. Currently, there are limited options to treat fungal infections, especially drug-resistant-fungal infections. Therefore, investigating alternative or repurposed antifungals is paramount. Endophytic microbes (EMs) and rhizospheric microbes (RMs) emerge as promising reservoirs of bioactive natural compounds. Interestingly, plants that have adapted to various environmental conditions harbour a plethora of microbes producing a variety of bioactive natural products that can be assessed for potential antifungal activity. To date, EMs and RMs residing in coastal plants and their associated antifungals have not been extensively studied or reviewed. Therefore, this comprehensive review will focus on antifungal natural products, extracted from coastal-vegetation-associated microbiota to draw the attention of research in this field. A comprehensive literature search was conducted by examining both Scopus and Google Scholar databases during the period of 2013-2024 related to the following coastal vegetation: mangroves, sand dune plants, salt marsh plants, and seagrasses. To date, 65 novel antifungal compounds derived from coastal-plant EMs and RMs have been identified. Mangroves were found to be the most prominent host harbouring antifungal-producing EMs and RMs compared with other coastal plants. Coastal-plant-associated fungal partners were the most prominent producers of antifungals compared to their bacterial counterparts. Fifty-four fungal-EM/RM derived antifungals have been reported to demonstrate activities against plant pathogenic fungi as well as human fungal pathogens. Most of the bacterial-derived antifungals (11 antifungals) have previously been reported to have antifungal activity against Candida albicans.

Invasive Candida bloodstream infections (candidemia) are a deadly global health threat. Rare Candida species are increasingly important causes of candidemia and phenotypic data, including patterns of antifungal drug resistance, is limited. There is geographic variation in the distribution of Candida species and frequency of antifungal drug resistance, which means that collecting and reporting regional data can have significant clinical value. Here, we report the first survey of species distribution, frequency of antifungal drug resistance, and phenotypic variability of Candida bloodstream isolates from an academic medical center and 5 affiliated hospitals in the Minneapolis-Saint Paul region of Minnesota, collected during an 18-month period from 2019 to 2021. We collected 288 isolates spanning 11 species from 119 patients. C. albicans was the most frequently recovered species, followed by C. glabrata and C. parapsilosis, with 10% of cases representing additional, rare species. We performed antifungal drug susceptibility for the three major drug classes and, concerningly, we identified fluconazole, micafungin and multidrug resistance rates in C. glabrata that were ~ 2 times higher than that reported in other regions of the United States. We report some of the first phenotypic data in rare non-albicans Candida species. Through analysis of serial isolates from individual patients, we identified clinically relevant within-patient differences of MIC values in multiple drug classes. Our results provide valuable clinical data relevant to antifungal stewardship efforts and highlight important areas of future research, including within-patient dynamics of infection and the mechanisms of drug resistance in rare Candida species.

Fungal infections constitute a significant global health threat, with an estimated incidence of 6.5 million invasive fungal infections and 2.5 million associated deaths each year. New antifungal agents are being developed to address the challenges of fungal infections management, driven by the evolving fungal epidemiology, the emergence of antifungal resistance, and the limitations of existing treatments.

This review provides a thorough overview of the latest developments in novel antifungal agents, highlighting pivotal evidence obtained from clinical trials.

New antifungal agents hold promising future for difficult-to-treat fungal infections, providing for improved bioavailability, pharmacokinetic properties, adverse events and drug interactions, as well as, spectrum of activity. However, further data is needed before incorporating these agents in everyday clinical practice for the management of invasive fungal infections.

The increasing detection rate of C. tropicalis and its azole resistance have made clinical treatment difficult. The presence of candiduria seems to correlate with invasive candida infection, especially for patients admitted to ICUs. However, the prevalence and antifungal resistance of C. tropicalis isolates in urine samples has not been well studied.

To retrospectively investigate the clinical features, antifungal resistance, and genetic relatedness of C. tropicalis isolates from urine samples.

A total of 107 clinical C. tropicalis isolates were retrospectively studied, including phenotypes of isolates and characteristics of patients. The genetic profiles of 107 isolates were genotyped using multi-locus sequence typing (MLST). Phylogenetic analysis was inferred using unweighted pair group method with arithmetic averages. MLST clonal clusters (CCs) were analysed by goeBURST.

Of the 107 isolates, 27.1% were resistant to fluconazole, and there was a notable increasing trend of fluconazole resistance from 16.1% in 2019 to 40.0% in 2021. Forty-seven diploid sequence types (DSTs) were assigned to ten major CCs. CC1 was the predominant fluconazole-susceptible group; 24 isolates from CC1 belonged to DST333, an outbreak clone in NICU ward. The azole-resistant CC4 contained 19 isolates, accounting for 65.5% of the azole-resistant isolates in this study. CC4 belongs to a prevalent FNS CC1 globally, of which the putative founder genotype was DST225.

This study revealed an outbreak of azole-susceptible C. tropicalis isolates in urine specimens and a high azole resistance rate of C. tropicalis in candiduria, and the MLST type showed clonal aggregation in azole-resistant isolates from urine samples.