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1.
Mycoses ; 67(7): e13757, 2024 Jul.
Article in English | MEDLINE | ID: mdl-39049157

ABSTRACT

BACKGROUND: Candida vulturna is an emerging pathogen belonging to the Metshnikowiaceae family together with Candida auris and Candida haemulonii species complex. Some strains of this species were reported to be resistant to several antifungal agents. OBJECTIVES: This study aims to address identification difficulties, evaluate antiungal susceptibilities and explore the molecular mechanisms of azole resistance of Candida vulturna. METHODS: We studied five C. vulturna clinical strains isolated in three Colombian cities. Identification was performed by phenotypical, proteomic and molecular methods. Antifungal susceptibility testing was performed following CLSI protocol. Its ERG11 genes were sequenced and a substitution was encountered in azole resistant isolates. To confirm the role of this substitution in the resistance phenotype, Saccharomyces cerevisiae strains with a chimeric ERG11 gene were created. RESULTS: Discrepancies in identification methods are highlighted. Sequencing confirmed the identification as C. vulturna. Antifungal susceptibility varied among strains, with four strains exhibiting reduced susceptibility to azoles and amphotericin B. ERG11 sequencing showed a point mutation (producing a P135S substitution) that was associated with the azole-resistant phenotype. CONCLUSIONS: This study contributes to the understanding of C. vulturna's identification challenges, its susceptibility patterns, and sheds light on its molecular mechanisms of azole resistance.


Subject(s)
Antifungal Agents , Azoles , Candida , Candidiasis , Microbial Sensitivity Tests , Antifungal Agents/pharmacology , Azoles/pharmacology , Candida/drug effects , Candida/genetics , Candida/classification , Candida/isolation & purification , Candidiasis/microbiology , Humans , Drug Resistance, Multiple, Fungal/genetics , Colombia , Amphotericin B/pharmacology , Drug Resistance, Fungal/genetics , Point Mutation , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/drug effects , Cytochrome P-450 Enzyme System/genetics , Fungal Proteins/genetics , Sequence Analysis, DNA , Saccharomyces cerevisiae Proteins
2.
Food Microbiol ; 123: 104584, 2024 Oct.
Article in English | MEDLINE | ID: mdl-39038890

ABSTRACT

A single strain of Candida anglica, isolated from cider, is available in international yeast collections. We present here seven new strains isolated from French PDO cheeses. For one of the cheese strains, we achieved a high-quality genome assembly of 13.7 Mb with eight near-complete telomere-to-telomere chromosomes. The genomes of two additional cheese strains and of the cider strain were also assembled and annotated, resulting in a core genome of 5966 coding sequences. Phylogenetic analysis showed that the seven cheese strains clustered together, away from the cider strain. Mating-type locus analysis revealed the presence of a MATa locus in the cider strain but a MATalpha locus in all cheese strains. The presence of LINE retrotransposons at identical genome position in the cheese strains, and two different karyotypic profiles resulting from chromosomal rearrangements were observed. Together, these findings are consistent with clonal propagation of the cheese strains. Phenotypic trait variations were observed within the cheese population under stress conditions whereas the cider strain was found to have a much greater capacity for growth in all conditions tested.


Subject(s)
Candida , Cheese , Fermented Foods , Genome, Fungal , Phylogeny , Cheese/microbiology , Candida/genetics , Candida/metabolism , Candida/classification , Candida/isolation & purification , Candida/growth & development , Fermented Foods/microbiology , Adaptation, Physiological , Food Microbiology , Fermentation , Genes, Mating Type, Fungal
3.
Microb Pathog ; 193: 106773, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38960213

ABSTRACT

Meyerozyma guilliermondii (Candida guilliermondii) is one of the Candida species associated with invasive candidiasis. With the potential for expressing industrially important enzymes, M. guilliermondii strain SO possessed 99 % proteome similarity with the clinical ATCC 6260 isolate and showed pathogenicity towards zebrafish embryos. Recently, three secreted aspartyl proteinases (SAPs) were computationally identified as potential virulence factors in this strain without in vitro verification of SAP activity. The quantification of Candida SAPs activity in liquid broth were also scarcely reported. Thus, this study aimed to characterize M. guilliermondii strain SO's ability to produce SAPs (MgSAPs) in different conditions (morphology and medium) besides analyzing its growth profile. MgSAPs' capability to cleave bovine serum albumin (BSA) was also determined to propose that MgSAPs as the potential virulence factors compared to the avirulent Saccharomyces cerevisiae. M. guilliermondii strain SO produced more SAPs (higher activity) in yeast nitrogen base-BSA-dextrose broth compared to yeast extract-BSA-dextrose broth despite insignificantly different SAP activity in both planktonic and biofilm cells. FeCl3 supplementation significantly increased the specific protein activity (∼40 %). The BSA cleavage by MgSAPs at an acidic pH was proven through semi-quantitative SDS-PAGE, sharing similar profile with HIV-1 retropepsin. The presented work highlighted the MgSAPs on fungal cell wall and extracellular milieu during host infection could be corroborated to the quantitative production in different growth modes presented herein besides shedding lights on the potential usage of retropepsin's inhibitors in treating candidiasis. Molecular and expression analyses of MgSAPs and their deletion should be further explored to attribute their respective virulence effects.


Subject(s)
Aspartic Acid Proteases , Biofilms , Candidiasis , Serum Albumin, Bovine , Virulence Factors , Virulence Factors/metabolism , Virulence Factors/genetics , Aspartic Acid Proteases/metabolism , Aspartic Acid Proteases/genetics , Candidiasis/microbiology , Serum Albumin, Bovine/metabolism , Biofilms/growth & development , Animals , Fungal Proteins/metabolism , Fungal Proteins/genetics , Culture Media/chemistry , Candida/pathogenicity , Candida/metabolism , Candida/genetics , Saccharomycetales/metabolism , Saccharomycetales/pathogenicity , Saccharomycetales/genetics , Virulence
4.
FEMS Yeast Res ; 242024 Jan 09.
Article in English | MEDLINE | ID: mdl-39009031

ABSTRACT

Lignocellulose (dry plant biomass) is an abundant cheap inedible residue of agriculture and wood industry with great potential as a feedstock for biotechnological processes. Lignocellulosic substrates can serve as valuable resources in fermentation processes, allowing the production of a wide array of chemicals, fuels, and food additives. The main obstacle for cost-effective conversion of lignocellulosic hydrolysates to target products is poor metabolism of the major pentoses, xylose and L-arabinose, which are the second and third most abundant sugars of lignocellulose after glucose. We study the oversynthesis of riboflavin in the flavinogenic yeast Candida famata and found that all major lignocellulosic sugars, including xylose and L-arabinose, support robust growth and riboflavin synthesis in the available strains of C. famata. To further increase riboflavin production from xylose and lignocellulose hydrolysate, genes XYL1 and XYL2 coding for xylose reductase and xylitol dehydrogenase were overexpressed. The resulting strains exhibited increased riboflavin production in both shake flasks and bioreactors using diluted hydrolysate, reaching 1.5 g L-1.


Subject(s)
Candida , Lignin , Metabolic Engineering , Riboflavin , Xylose , Lignin/metabolism , Riboflavin/metabolism , Riboflavin/biosynthesis , Candida/metabolism , Candida/genetics , Xylose/metabolism , Aldehyde Reductase/metabolism , Aldehyde Reductase/genetics , Fermentation , Bioreactors/microbiology , D-Xylulose Reductase/metabolism , D-Xylulose Reductase/genetics , Arabinose/metabolism
5.
Sci Total Environ ; 947: 174662, 2024 Oct 15.
Article in English | MEDLINE | ID: mdl-38997029

ABSTRACT

The use of recreational waters is a widespread activity worldwide, and one of the risks associated with this practice is the exposure of bathers to microorganisms that may arise due to pollution caused by inadequate infrastructure and sanitation. In the present work, we isolated Candida spp. (n = 24) from five recreational beaches in Rio de Janeiro, Brazil, in order to evaluate their susceptibility to antifungals, the production of virulence attributes and the in vivo virulence using Tenebrio molitor larvae as a model. The ITS1-5.8S-ITS2 gene sequencing identified thirteen isolates (54.1 %) as C. tropicalis, seven (29.1 %) as C. krusei (Pichia kudriavzevii), one (4.2 %) as C. rugosa (Diutina rugosa), one (4.2 %) as C. mesorugosa (Diutina mesorugosa), one (4.2 %) as C. utilis (Cyberlindnera jadinii) and one (4.2 %) as C. parapsilosis. C. tropicalis isolates showed resistance to azoles and susceptibility to amphotericin B, flucytosine and caspofungin. C. krusei isolates were resistant to fluconazole, caspofungin and itraconazole, with 42.8 % resistance to flucytosine, besides susceptibility to voriconazole and amphotericin B. The remaining species were susceptible to all tested antifungals. All Candida isolates adhered to abiotic surfaces and formed biofilm on polystyrene, albeit to varying degrees, and produced aspartic protease and hemolytic activity, which are considered fungal virulence attributes. C. tropicalis, C. krusei and C. utilis isolates produced phytase, while the only esterase producer was C. tropicalis. Regarding resistance to osmotic stress, all isolates of C. tropicalis, C. parapsilosis and C. mesorugosa grew up to 7.5 % NaCl; the remaining isolates grew up to 1.87-3.75 % NaCl. The mortality caused by fungal challenges in T. molitor larvae was variable, with C. tropicalis, C. utilis and C. parapsilosis being more virulent than C. krusei and C. rugosa complex. Collectively, the presence of these yeasts, particularly the virulent and resistant isolates, in recreational waters can pose a significant health risk to bathers.


Subject(s)
Antifungal Agents , Candida , Drug Resistance, Fungal , Brazil , Antifungal Agents/pharmacology , Candida/drug effects , Candida/pathogenicity , Candida/genetics , Virulence , Microbial Sensitivity Tests , Animals , Bathing Beaches
6.
BMC Biotechnol ; 24(1): 43, 2024 Jun 22.
Article in English | MEDLINE | ID: mdl-38909197

ABSTRACT

Fungal diseases are often linked to poverty, which is associated with poor hygiene and sanitation conditions that have been severely worsened by the COVID-19 pandemic. Moreover, COVID-19 patients are treated with Dexamethasone, a corticosteroid that promotes an immunosuppressive profile, making patients more susceptible to opportunistic fungal infections, such as those caused by Candida species. In this study, we analyzed the prevalence of Candida yeasts in wastewater samples collected to track viral genetic material during the COVID-19 pandemic and identified the yeasts using polyphasic taxonomy. Furthermore, we investigated the production of biofilm and hydrolytic enzymes, which are known virulence factors. Our findings revealed that all Candida species could form biofilms and exhibited moderate hydrolytic enzyme activity. We also proposed a workflow for monitoring wastewater using Colony PCR instead of conventional PCR, as this technique is fast, cost-effective, and reliable. This approach enhances the accurate taxonomic identification of yeasts in environmental samples, contributing to environmental monitoring as part of the One Health approach, which preconizes the monitoring of possible emergent pathogenic microorganisms, including fungi.


Subject(s)
COVID-19 , Candida , Wastewater , Workflow , Wastewater/microbiology , Wastewater/virology , Brazil/epidemiology , Candida/isolation & purification , Candida/genetics , Candida/classification , COVID-19/epidemiology , COVID-19/virology , Humans , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Biofilms , Environmental Monitoring/methods , Pandemics
7.
Microbiol Spectr ; 12(7): e0354023, 2024 Jul 02.
Article in English | MEDLINE | ID: mdl-38842332

ABSTRACT

Candida auris, initially identified in 2009, has rapidly become a critical concern due to its antifungal resistance and significant mortality rates in healthcare-associated outbreaks. To date, whole-genome sequencing (WGS) has identified five unique clades of C. auris, with some strains displaying resistance to all primary antifungal drug classes. In this study, we presented the first WGS analysis of C. auris from Bangladesh, describing its origins, transmission dynamics, and antifungal susceptibility testing (AFST) profile. Ten C. auris isolates collected from hospital settings in Bangladesh were initially identified by CHROMagar Candida Plus, followed by VITEK2 system, and later sequenced using Illumina NextSeq 550 system. Reference-based phylogenetic analysis and variant calling pipelines were used to classify the isolates in different clades. All isolates aligned ~90% with the Clade I C. auris B11205 reference genome. Of the 10 isolates, 8 were clustered with Clade I isolates, highlighting a South Asian lineage prevalent in Bangladesh. Remarkably, the remaining two isolates formed a distinct cluster, exhibiting >42,447 single-nucleotide polymorphism differences compared to their closest Clade IV counterparts. This significant variation corroborates the emergence of a sixth clade (Clade VI) of C. auris in Bangladesh, with potential for international transmission. AFST results showed that 80% of the C. auris isolates were resistant to fluconazole and voriconazole, whereas Clade VI isolates were susceptible to azoles, echinocandins, and pyrimidine analogue. Genomic sequencing revealed ERG11_Y132F mutation conferring azole resistance while FCY1_S70R mutation found inconsequential in describing 5-flucytosine resistance. Our study underscores the pressing need for comprehensive genomic surveillance in Bangladesh to better understand the emergence, transmission dynamics, and resistance profiles of C. auris infections. Unveiling the discovery of a sixth clade (Clade VI) accentuates the indispensable role of advanced sequencing methodologies.IMPORTANCECandida auris is a nosocomial fungal pathogen that is commonly misidentified as other Candida species. Since its emergence in 2009, this multidrug-resistant fungus has become one of the five urgent antimicrobial threats by 2019. Whole-genome sequencing (WGS) has proven to be the most accurate identification technique of C. auris which also played a crucial role in the initial discovery of this pathogen. WGS analysis of C. auris has revealed five distinct clades where isolates of each clade differ among themselves based on pathogenicity, colonization, infection mechanism, as well as other phenotypic characteristics. In Bangladesh, C. auris was first reported in 2019 from clinical samples of a large hospital in Dhaka city. To understand the origin, transmission dynamics, and antifungal-resistance profile of C. auris isolates circulating in Bangladesh, we conducted a WGS-based surveillance study on two of the largest hospital settings in Dhaka, Bangladesh.


Subject(s)
Antifungal Agents , Candida auris , Candidiasis , Microbial Sensitivity Tests , Phylogeny , Whole Genome Sequencing , Bangladesh/epidemiology , Humans , Antifungal Agents/pharmacology , Candidiasis/microbiology , Candidiasis/epidemiology , Candida auris/genetics , Candida auris/drug effects , Candida auris/isolation & purification , Drug Resistance, Fungal , Genome, Fungal , Polymorphism, Single Nucleotide , Candida/genetics , Candida/drug effects , Candida/classification , Candida/isolation & purification , Fluconazole/pharmacology , Female
8.
Med Mycol ; 62(7)2024 Jul 04.
Article in English | MEDLINE | ID: mdl-38877671

ABSTRACT

Candida auris is an emerging fungal pathogen associated with multi-drug resistance rates and widespread outbreaks in hospitals and healthcare units worldwide. Sequencing studies have revealed that different clonal lineages of the fungus seem to be prevalent among distinct geographical sites. The first case of C. auris in Northern Greece was reported in Thessaloniki in October 2022, almost 2 years after the first isolation in Greece (Athens 2019). The Mycology Laboratory of the Medical School of Aristotle University of Thessaloniki stands as the reference laboratory for fungal diseases in Northern Greece and a meticulous search for the yeast, in plenty of suspicious samples, has been run since 2019 in the Lab as well as a retrospective analysis of all its yeasts' collection, back to 2008, with negative results for the presence of C. auris. Here, are presented the findings concerning the outbreak and surveillance of C. auris in Northern Greece, mainly the region of Thessaloniki and the broader area of Macedonia, from October 2022 until August 2023. The isolates from Northern Greece continue to fall in Clade I and present with an almost equal and stable sensitivity profile until now.


The study concerns the outbreak of Candida auris in Northern Greece since October 2022 and the effort for surveillance and epidemiological monitoring. All isolates continue to fall in Clade I and present with an almost equal and stable sensitivity profile till now.


Subject(s)
Candida auris , Candidiasis , Disease Outbreaks , Epidemiological Monitoring , Greece/epidemiology , Humans , Candidiasis/epidemiology , Candidiasis/microbiology , Candida auris/genetics , Candida auris/isolation & purification , Retrospective Studies , Antifungal Agents/pharmacology , Microbial Sensitivity Tests , Male , Drug Resistance, Multiple, Fungal , Candida/isolation & purification , Candida/classification , Candida/genetics , Female
9.
J Clin Microbiol ; 62(7): e0052524, 2024 Jul 16.
Article in English | MEDLINE | ID: mdl-38888304

ABSTRACT

Candida auris is a multidrug-resistant fungal pathogen with a propensity to colonize humans and persist on environmental surfaces. C. auris invasive fungal disease is being increasingly identified in acute and long-term care settings. We have developed a prototype cartridge-based C. auris surveillance assay (CaurisSurV cartridge; "research use only") that includes integrated sample processing and nucleic acid amplification to detect C. auris from surveillance skin swabs in the GeneXpert instrument and is designed for point-of-care use. The assay limit of detection (LoD) in the skin swab matrix was 10.5 and 14.8 CFU/mL for non-aggregative (AR0388) and aggregative (AR0382) strains of C. auris, respectively. All five known clades of C. auris were detected at 2-3-5× (31.5-52.5 CFU/mL) the LoD. The assay was validated using a total of 85 clinical swab samples banked at two different institutions (University of California Los Angeles, CA and Wadsworth Center, NY). Compared to culture, sensitivity was 96.8% (30/31) and 100% (10/10) in the UCLA and Wadsworth cohorts, respectively, providing a combined sensitivity of 97.5% (40/41), and compared to PCR, the combined sensitivity was 92% (46/50). Specificity was 100% with both clinical (C. auris negative matrix, N = 31) and analytical (non-C. auris strains, N = 32) samples. An additional blinded study with N = 60 samples from Wadsworth Center, NY yielded 97% (29/30) sensitivity and 100% (28/28) specificity. We have developed a completely integrated, sensitive, specific, and 58-min prototype test, which can be used for routine surveillance of C. auris and might help prevent colonization and outbreaks in acute and chronic healthcare settings. IMPORTANCE: This study has the potential to offer a better solution to healthcare providers at hospitals and long-term care facilities in their ongoing efforts for effective and timely control of Candida auris infection and hence quicker response for any potential future outbreaks.


Subject(s)
Candida auris , Candidiasis , Sensitivity and Specificity , Humans , Candidiasis/diagnosis , Candidiasis/microbiology , Candida auris/genetics , Infection Control/methods , Epidemiological Monitoring , Skin/microbiology , Limit of Detection , Point-of-Care Systems , Nucleic Acid Amplification Techniques/methods , Molecular Diagnostic Techniques/methods , Candida/isolation & purification , Candida/genetics , Candida/classification
10.
J Microbiol Biotechnol ; 34(7): 1365-1375, 2024 Jul 28.
Article in English | MEDLINE | ID: mdl-38881183

ABSTRACT

The rise of Candida auris, a multidrug-resistant fungal pathogen, across more than 40 countries, has signaled an alarming threat to global health due to its significant resistance to existing antifungal therapies. Characterized by its rapid spread and robust drug resistance, C. auris presents a critical challenge in managing infections, particularly in healthcare settings. With research on its biological traits and genetic basis of virulence and resistance still in the early stages, there is a pressing need for a concerted effort to understand and counteract this pathogen. This review synthesizes current knowledge on the epidemiology, biology, genetic manipulation, pathogenicity, diagnostics, and resistance mechanisms of C. auris, and discusses future directions in research and therapeutic development. By exploring the complexities surrounding C. auris, we aim to underscore the importance of advancing research to devise effective control and treatment strategies.


Subject(s)
Antifungal Agents , Candida auris , Candidiasis , Drug Resistance, Multiple, Fungal , Humans , Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use , Drug Resistance, Multiple, Fungal/genetics , Candidiasis/microbiology , Candidiasis/drug therapy , Candida auris/genetics , Candida auris/drug effects , Virulence , Animals , Candida/drug effects , Candida/genetics , Candida/pathogenicity
12.
Arch Microbiol ; 206(6): 255, 2024 May 11.
Article in English | MEDLINE | ID: mdl-38734793

ABSTRACT

Cystic fibrosis (CF) is an inherited disease that results from mutations in the gene responsible for the cystic fibrosis transmembrane conductance regulator (CFTR). The airways become clogged with thick, viscous mucus that traps microbes in respiratory tracts, facilitating colonization, inflammation and infection. CF is recognized as a biofilm-associated disease, it is commonly polymicrobial and can develop in biofilms. This review discusses Candida spp. and both Gram-positive and Gram-negative bacterial biofilms that affect the airways and cause pulmonary infections in the CF context, with a particular focus on mixed-species biofilms. In addition, the review explores the intricate interactions between fungal and bacterial species within these biofilms and elucidates the underlying molecular mechanisms that govern their dynamics. Moreover, the review addresses the multifaceted issue of antimicrobial resistance in the context of CF-associated biofilms. By synthesizing current knowledge and research findings, this review aims to provide insights into the pathogenesis of CF-related infections and identify potential therapeutic approaches to manage and combat these complex biofilm-mediated infections.


Subject(s)
Biofilms , Candida , Cystic Fibrosis , Biofilms/growth & development , Cystic Fibrosis/microbiology , Humans , Candida/physiology , Candida/genetics , Candidiasis/microbiology , Gram-Negative Bacteria/physiology , Gram-Negative Bacteria/genetics , Anti-Bacterial Agents/pharmacology
13.
J Mycol Med ; 34(2): 101482, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38763122

ABSTRACT

Fungal infections in neonatal intensive care units (NICU) are mainly related to Candida species, with high mortality rates. They are predominantly of endogenous origin, however, cross-infection transmitted by healthcare professionals' hands has occurred. The aim of this study was to identify Candida species isolated from the hands of healthcare professionals in a NICU before and after hygiene with 70% ethanol-based gel and evaluate virulence factors DNase, phospholipase, proteinase, hemolysin, biofilm biomass production, and metabolic activity. In vitro antifungal susceptibility testing and similarity by random amplified polymorphic DNA (RAPD) were also performed. C. parapsilosis complex was the most frequent species (57.1%); all isolates presented at least one virulence factor; three isolates (Candida parapsilosis complex) were resistant to amphotericin B, two (Candida famata [currently Debaryomyces hansenii] and Candida guilliermondii [currently Meyerozyma guilliermondii]) was resistant to micafungin, and six (Candida parapsilosis complex, Candida guilliermondii [=Meyerozyma guilliermondii], Candida viswanathi, Candida catenulata [currently Diutina catenulata] and Candida lusitaniae [currently Clavispora lusitaniae]) were resistant to fluconazole. Molecular analysis by RAPD revealed two clusters of identical strains that were in the hands of distinct professionals. Candida spp. were isolated even after hygiene with 70% ethanol-based gel, highlighting the importance of stricter basic measures for hospital infection control to prevent nosocomial transmission.


Subject(s)
Antifungal Agents , Candida , Cross Infection , Ethanol , Hand , Microbial Sensitivity Tests , Virulence Factors , Humans , Hand/microbiology , Antifungal Agents/pharmacology , Virulence Factors/genetics , Candida/drug effects , Candida/isolation & purification , Candida/genetics , Candida/pathogenicity , Ethanol/pharmacology , Cross Infection/microbiology , Cross Infection/prevention & control , Candidiasis/microbiology , Health Personnel , Random Amplified Polymorphic DNA Technique , Biofilms/drug effects , Biofilms/growth & development , Intensive Care Units, Neonatal , Drug Resistance, Fungal , Gels , Hand Disinfection
14.
ACS Synth Biol ; 13(6): 1716-1726, 2024 Jun 21.
Article in English | MEDLINE | ID: mdl-38733342

ABSTRACT

NAD is a redox coenzyme and is the center of energy metabolism. In metabolic engineering modifications, an insufficient NAD(H) supply often limits the accumulation of target products. In this study, Candida glycerinogenes was found to be able to supply NAD(H) in large fluxes, up to 7.6 times more than Saccharomyces cerevisiae in aerobic fermentation. Aerobic fermentation in a medium without amino nitrogen sources demonstrated that C. glycerinogenes NAD synthesis was not dependent on NAD precursors in the medium. Inhibition by antisense RNA and the detection of transcript levels indicated that the main NAD supply pathway is the de novo biosynthesis pathway. It was further demonstrated that NAD(H) supply was unaffected by changes in metabolic flow through C. glycerinogenes ΔGPD aerobic fermentation (80 g/L ethanol). In conclusion, the ability of C. glycerinogenes to supply NAD(H) in large fluxes provides a new approach to solving the NAD(H) supply problem in synthetic biology.


Subject(s)
Candida , Fermentation , Metabolic Engineering , NAD , NAD/metabolism , Candida/metabolism , Candida/genetics , Aerobiosis , Metabolic Engineering/methods , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae/genetics , Ethanol/metabolism , RNA, Antisense/genetics , RNA, Antisense/metabolism
15.
J Clin Microbiol ; 62(6): e0015824, 2024 Jun 12.
Article in English | MEDLINE | ID: mdl-38690882

ABSTRACT

Candida auris is a multidrug-resistant opportunistic fungal pathogen capable of causing serious infections and healthcare-associated outbreaks. Screening for colonization with C. auris has become routine and is recommended in many hospitals and healthcare facilities as an infection control and prevention strategy. Subsequently, and since there are currently no FDA-approved tests for this purpose, clinical microbiology laboratories have become responsible for developing protocols to detect C. auris using axial and inguinal screening swabs. In a College of American Pathologists-accredited large academic healthcare center setting, we implemented a laboratory-developed nucleic-acid amplification test for the detection of C. auris DNA. Our test validation evaluated the performance of the DiaSorin C. auris primer set used in a real-time qualitative PCR assay on the LIAISON MDX thermocycler with the Simplexa Universal Disc. The assay was highly sensitive and specific, with a limit of detection of 1-2 CFU/reaction, with no observed cross-reactivity with other Candida spp., bacterial skin commensal organisms or commonly encountered viruses. When run in parallel with a culture-based detection method, the PCR assay was 100% sensitive and specific. The assay was precise, with low variability between replicates within and between runs. Lastly, pre-analytical factors, including swab storage time, temperature, and transport media, were assessed and found to have no significant effect on the detection of C. auris at variable concentrations. Taken together, this study expands the available options for nucleic acid detection of C. auris and characterizes pre-analytical factors for implementation in both high- and low-volume laboratory settings. IMPORTANCE: This study overviews the validation and implementation of a molecular screening tool for the detection of Candida auris in a College of American Pathologist-accredited clinical laboratory. This molecular laboratory-developed test is both highly sensitive and specific and has significant health-system cost-savings associated with significantly reduced turn-around-time compared to traditional standard-of-care culture-based work up. This method and workflow is of interest to support clinical microbiology diagnostics and to help aid in hospital inpatient, and infection prevention control screening.


Subject(s)
Candida auris , Candidiasis , Real-Time Polymerase Chain Reaction , Sensitivity and Specificity , Humans , Real-Time Polymerase Chain Reaction/methods , Candidiasis/diagnosis , Candidiasis/microbiology , Candida auris/genetics , Mass Screening/methods , Inpatients , Molecular Diagnostic Techniques/methods , Molecular Diagnostic Techniques/standards , Hospitals , Candida/genetics , Candida/isolation & purification , DNA, Fungal/genetics
16.
Med Mycol ; 62(7)2024 Jul 04.
Article in English | MEDLINE | ID: mdl-38816207

ABSTRACT

Candida auris represents one of the most urgent threats to public health, although its ecology remains largely unknown. Because amphibians and reptiles may present favorable conditions for C. auris colonization, cloacal and blood samples (n = 68), from several snake species, were cultured and molecularly screened for C. auris using molecular amplification of glycosylphosphatidylinositol protein-encoding genes and ribosomal internal transcribed spacer sequencing. Candida auris was isolated from the cloacal swab of one Egyptian cobra (Naja haje legionis) and molecularly identified in its cloaca and blood. The isolation of C. auris from wild animals is herein reported for the first time, thus suggesting the role that these animals could play as reservoirs of this emerging pathogen. The occurrence of C. auris in blood requires further investigation, although the presence of cationic antimicrobial peptides in the plasma of reptiles could play a role in reducing the vitality of the fungus.


Candida auris represents one of the most urgent threats to public health. In this study, we reported for the first time the isolation of C. auris from snake thus suggesting the role of these animals as reservoirs of this emerging pathogen.


Subject(s)
Candida , Candidiasis , DNA, Ribosomal Spacer , Disease Reservoirs , Animals , Candida/genetics , Candida/classification , Candida/isolation & purification , Candida/drug effects , Disease Reservoirs/microbiology , Candidiasis/microbiology , Candidiasis/veterinary , DNA, Ribosomal Spacer/genetics , DNA, Ribosomal Spacer/chemistry , Cloaca/microbiology , Sequence Analysis, DNA , DNA, Fungal/genetics , Blood/microbiology , Snakes/microbiology , Elapidae , Egypt , Phylogeny
17.
Med Mycol ; 62(7)2024 Jul 04.
Article in English | MEDLINE | ID: mdl-38806253

ABSTRACT

Candida lusitaniae fungemia is a serious infection that is rarely reported in children. The aim of this study is to describe a case series of C. lusitaniae fungemia and review previous publications regarding this rare pathogen. This is a multicenter case series of children diagnosed with C. lusitaniae fungemia. A total of 18 cases that occurred over a 15-year period in five tertiary hospitals were included. Additionally, a review of the literature regarding C. lusitaniae fungemia in children was performed. A total of 18 cases were enrolled; 11/18 (61%) were males, with a mean age of 2.3 years. All patients had severe underlying diseases and risk factors for opportunistic infection, most commonly prematurity and malignancies. More than one-third of cases occurred during the last 2 years of the study period. All isolates were susceptible to all tested antifungals. The survival rate following the acute infection was 94%, whereas the survival rate of 14 previously published cases was 71%, with the most common underlying diseases being CGD and malignancies. Candida lusitaniae fungemia is not a common event in the pediatric population, occurring exclusively in children with severe underlying diseases and significant risk factors. This cohort revealed better clinical outcomes than previously reported. All tested isolates were susceptible to all antifungal agents; variability in susceptibility as previously reported was not found in this study. The allegedly higher rate of infection in recent years is in need of further investigation in larger prospective studies in order to conclude if a real trend is at play.


Candida lusitaniae fungemia is a serious infection rarely reported in children. This cohort revealed better clinical outcomes than previously reported. All tested isolates were susceptible to all antifungal agents. The higher rate of infection in recent years is in need of further investigation.


Subject(s)
Antifungal Agents , Candida , Child, Preschool , Female , Humans , Male , Antifungal Agents/therapeutic use , Antifungal Agents/pharmacology , Candida/drug effects , Candida/genetics , Candida/isolation & purification , Candida/pathogenicity , Candidemia/microbiology , Candidemia/epidemiology , Fungemia/microbiology , Fungemia/mortality , Microbial Sensitivity Tests , Retrospective Studies , Risk Factors , Tertiary Care Centers/statistics & numerical data
18.
Front Cell Infect Microbiol ; 14: 1295841, 2024.
Article in English | MEDLINE | ID: mdl-38707510

ABSTRACT

Introduction: Although the existence of Candida species in the respiratory tract is often considered commensal, it is crucial to recognize the significance of Candida colonization in immunocompromised or COVID-19 patients. The emergence of Candida auris as an emerging pathogen further emphasizes the importance of monitoring yeast infection/colonization, particularly in COVID-19 patients. Methods: In this study, respiratory samples mainly from COVID-19 patients, primarily those suspected of having a fungal infection, were cultured on Sabouraud dextrose agar plates and the yeast colonies were identified using a two-step multiplex PCR method. The samples suspected of C. auris underwent specific nested PCR followed by sequence analysis. Results: A total of 199 respiratory samples were collected from 73 women and 126 men, ranging in age from 1.6 to 88 years. Among the patients, 141 had COVID-19, 32 had cancer, 5 were hospitalized in ICU, 2 had chronic obstructive pulmonary disease)COPD(, and others were patients with combination diseases. From these samples, a total of 334 yeast strains were identified. C. albicans (n=132, 39.52%) was the most common species, followed by C. tropicalis (n=67, 20%), C. glabrata (n=56, 16.76%), C. krusei (n=18, 5.4%), C. parapsilosis (n=17, 5.08%), Saccharomyces cerevisiae (n=10, 3%), C. kefyr (n=9, 2.6%), C. dubliniensis (n=7, 2.1%), C. lusitaniae (n=5, 1.5%), C. auris (n=3, 0.9%), C. guilliermondii (n=2, 0.6%), C. rugosa (n=1, 0.3%), C. intermedia (n=1, 0.3%), and Trichosporon spp. (n=1, 0.3%). C. auris was detected in a patient in ICU and two COVID-19 patients. While its presence was confirmed through sequence analysis, our extensive efforts to isolate C. auris were unsuccessful. Conclusion: While C. albicans colonization remains prevalent, our study found no evidence of Candida lung infection. Since the role of Candida colonization in airway secretions remains ambiguous due to limited research, further studies are imperative to shed light on this matter.


Subject(s)
COVID-19 , Candida auris , Candidiasis , SARS-CoV-2 , Humans , COVID-19/microbiology , Aged , Middle Aged , Female , Male , Aged, 80 and over , Adult , Child, Preschool , Candidiasis/microbiology , Child , Adolescent , Young Adult , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Infant , Candida auris/genetics , Candida auris/isolation & purification , Candida/isolation & purification , Candida/classification , Candida/genetics , Respiratory System/microbiology , Respiratory System/virology , Multiplex Polymerase Chain Reaction
19.
Sci Rep ; 14(1): 11597, 2024 05 21.
Article in English | MEDLINE | ID: mdl-38773138

ABSTRACT

Candida is the most prevalent fungal bloodstream infection (BSI) with a high mortality rate among hospitalized patients. Another concern facing physicians is rising global incidence of drug-resistant Candida. This study aimed to characterize the prevalence, antifungal susceptibility, biofilm formation, and virulence genes (HWP1, ALS1, SAP2) of different Candida spp. isolated from patients with candidemia. 52 isolates of Candida spp. were identified from blood cultures by chromogenic Candida agar and confirmed by the VITEK 2 system. Isolates were tested for antifungal susceptibility by disk diffusion and VITEK 2 system. Biofilm formation and investigated genes were detected by the Congo red method and conventional PCR, respectively. Candida spp. caused 2.3% of detected BSIs, of which 32.7% were caused by Candida albicans (C. albicans) and 67.3% by non-albicans Candida (NAC), with the predominance of C. tropicalis (25%), followed by C. parapsilosis (17.3%), and C. krusei (13.5%). The susceptibility rates to fluconazole, voriconazole, caspofungin, micafungin, amphotericin B, and flucytosine were 64.7%, 76.5%, 100.0%, 100%, 100.0%, and 100.0% in C. albicans, while 53.6%, 71.4%, 91.4%, 91.4%, 94.3%, and 94.3% in NAC, respectively. Biofilm production, HWP1, ALS1, and SAP2 were detected in 70.6%, 82.4%, 76.5%, and 52.9% of C. albicans and 74.3%, 85.7%, 80.0%, and 48.6% of NAC, respectively. There is remarkable shift to NAC BSIs and high azole resistance. Antifungal stewardship and analysis of risk factors associated with this shift are needed.


Subject(s)
Antifungal Agents , Biofilms , Candida , Candidemia , Drug Resistance, Fungal , Microbial Sensitivity Tests , Humans , Candidemia/microbiology , Candidemia/drug therapy , Candidemia/epidemiology , Antifungal Agents/pharmacology , Biofilms/drug effects , Biofilms/growth & development , Candida/drug effects , Candida/isolation & purification , Candida/pathogenicity , Candida/genetics , Virulence Factors/genetics , Virulence , Female , Male , Middle Aged , Adult
20.
Mycopathologia ; 189(3): 32, 2024 Apr 15.
Article in English | MEDLINE | ID: mdl-38622365

ABSTRACT

The rare fungus Candida saopaulonensis has never been reported to be associated with human infection. We report the draft genome sequence of the first clinical isolate of C. saopaulonensis, which was isolated from a very premature infant with sepsis. This is the first genome assembly reaching the near-complete chromosomal level with structural annotation for this species, opening up avenues for exploring evolutionary patterns and genetic mechanisms of pathogenesis.


Subject(s)
Candida , Sepsis , Humans , Infant, Newborn , Candida/genetics , Genome, Fungal , Infant, Premature
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