Characterization of Virulence Factors in Candida Species Causing Candidemia in a Tertiary Care Hospital in Bangkok, Thailand

Candidemia is often associated with high mortality, and Candida albicans, Candida tropicalis, Candida glabrata, and Candida parapsilosis are common causes of this disease. The pathogenicity characteristics of specific Candida spp. that cause candidemia in Thailand are poorly understood. This study aimed to characterize the virulence factors of Candida spp. Thirty-eight isolates of different Candida species from blood cultures were evaluated for their virulence properties, including exoenzyme and biofilm production, cell surface hydrophobicity, tissue invasion, epithelial cell damage, morphogenesis, and phagocytosis resistance; the identity and frequency of mutations in ERG11 contributing to azole-resistance were also determined. C. albicans had the highest epithelial cell invasion rate and phospholipase activity, with true hyphae formation, whereas C. tropicalis produced the most biofilm, hydrophobicity, protease activity, and host cell damage and true hyphae formation. ERG11 mutations Y132F and S154F were observed in all azole-resistant C. tropicalis. C. glabrata had the most hemolytic activity while cell invasion was low with no morphologic transition. C. glabrata was more easily phagocytosed than other species. C. parapsilosis generated pseudohyphae but not hyphae and did not exhibit any trends in exoenzyme production. This knowledge will be crucial for understanding the pathogenicity of Candida spp. and will help to explore antivirulence-based treatment.

[1]  S. Paul,et al.  Mechanisms of azole antifungal resistance in clinical isolates of Candida tropicalis , 2022, PloS one.

[2]  T. Wongsuk,et al.  Species Distribution, Antifungal Susceptibility, and Molecular Epidemiology of Candida Species Causing Candidemia in a Tertiary Care Hospital in Bangkok, Thailand , 2021, Journal of fungi.

[3]  M. Chayakulkeeree,et al.  Risk Factors and Outcomes of Non-albicans Candida Bloodstream Infection in Patients with Candidemia at Siriraj Hospital—Thailand’s Largest National Tertiary Referral Hospital , 2021, Journal of fungi.

[4]  K. Zomorodian,et al.  High detection of virulence factors by Candida species isolated from bloodstream of patients with candidemia. , 2020, Microbial pathogenesis.

[5]  T. West,et al.  Genomic loss in environmental and isogenic morphotype isolates of Burkholderia pseudomallei is associated with intracellular survival and plaque-forming efficiency , 2020, PLoS neglected tropical diseases.

[6]  P. Chongtrakool,et al.  An Association of an eBURST Group With Triazole Resistance of Candida tropicalis Blood Isolates , 2020, Frontiers in Microbiology.

[7]  Duncan W. Wilson,et al.  Cooperative Role of MAPK Pathways in the Interaction of Candida albicans with the Host Epithelium , 2019, Microorganisms.

[8]  A. Kummasook,et al.  Anti-fungal susceptibility and virulence factors of Candida spp. isolated from blood cultures. , 2019, Journal de mycologie medicale.

[9]  N. Soliman,et al.  Prevalence of Candida blood stream infections among children in tertiary care hospital: detection of species and antifungal susceptibility , 2019, Infection and drug resistance.

[10]  S. Paul,et al.  Extensive ERG11 mutations associated with fluconazole-resistant Candida albicans isolated from HIV-infected patients , 2019, Current medical mycology.

[11]  J. Jayaweera,et al.  The emergence of non-albicans candidemia and evaluation of HiChrome Candida differential agar and VITEK2 YST® platform for differentiation of Candida bloodstream isolates in teaching hospital Kandy, Sri Lanka , 2019, BMC Microbiology.

[12]  J. Jayaweera,et al.  The emergence of non-albicans candidemia and evaluation of HiChrome Candida differential agar and VITEK2 YST® platform for differentiation of Candida bloodstream isolates in teaching hospital Kandy, Sri Lanka , 2019, BMC Microbiology.

[13]  Y. An,et al.  Epidemiology, species distribution, antifungal susceptibility and mortality risk factors of candidemia among critically ill patients: a retrospective study from 2011 to 2017 in a teaching hospital in China , 2019, Antimicrobial resistance and infection control.

[14]  L. C. D. Souza,et al.  Virulence factors of Candida spp. obtained from blood cultures of patients with candidemia attended at tertiary hospitals in Northeast Brazil. , 2019, Journal de mycologie medicale.

[15]  M. Shams-Ghahfarokhi,et al.  Comparative analysis of proteinase, phospholipase, hydrophobicity and biofilm forming ability in Candida species isolated from clinical specimens. , 2018, Journal de mycologie medicale.

[16]  Z. Dagher,et al.  Fluorescent Tracking of Yeast Division Clarifies the Essential Role of Spleen Tyrosine Kinase in the Intracellular Control of Candida glabrata in Macrophages , 2018, Front. Immunol..

[17]  Qi Wang,et al.  MDR1 overexpression combined with ERG11 mutations induce high-level fluconazole resistance in Candida tropicalis clinical isolates , 2018, BMC Infectious Diseases.

[18]  Miguel C. Teixeira,et al.  Candida Biofilms: Threats, Challenges, and Promising Strategies , 2018, Front. Med..

[19]  M. E. S. Ferreira,et al.  Prevalence, virulence factors and antifungal susceptibility of Candida spp. isolated from bloodstream infections in a tertiary care hospital in Brazil , 2018, Mycoses.

[20]  G. M. Chaves,et al.  An Update on Candida tropicalis Based on Basic and Clinical Approaches , 2017, Front. Microbiol..

[21]  A. Ayadi,et al.  Virulence factors, antifungal susceptibility and molecular mechanisms of azole resistance among Candida parapsilosis complex isolates recovered from clinical specimens , 2017, Journal of Biomedical Science.

[22]  J. Teo,et al.  ERG11 mutations are associated with high-level azole resistance in clinical Candida tropicalis isolates, a Singapore study , 2017 .

[23]  E. Berkow,et al.  Azole Antifungal Resistance in Candida albicans and Emerging Non-albicans Candida Species , 2017, Frontiers in microbiology.

[24]  M. Kaur,et al.  Studying the Prevalence, Species Distribution, and Detection of In vitro Production of Phospholipase from Candida Isolated from Cases of Invasive Candidiasis , 2017, Journal of global infectious diseases.

[25]  Zhixin Liang,et al.  Nosocomial Bloodstream Infection Due to Candida spp. in China: Species Distribution, Clinical Features, and Outcomes , 2016, Mycopathologia.

[26]  A. Marra,et al.  Epidemiology and Microbiologic Characterization of Nosocomial Candidemia from a Brazilian National Surveillance Program , 2016, PloS one.

[27]  E. Berk,et al.  Investigation of the relationship between virulence factors and genotype of Candida spp. isolated from blood cultures. , 2015, Journal of infection in developing countries.

[28]  Wei Liu,et al.  The A395T Mutation in ERG11 Gene Confers Fluconazole Resistance in Candida tropicalis Causing Candidemia , 2015, Mycopathologia.

[29]  A. Rodrigues,et al.  Adhesion, biofilm formation, cell surface hydrophobicity, and antifungal planktonic susceptibility: relationship among Candida spp. , 2015, Front. Microbiol..

[30]  D. Moyes,et al.  Candida albicans-epithelial interactions and pathogenicity mechanisms: scratching the surface , 2015, Virulence.

[31]  M. C. Furlaneto,et al.  Effects of human blood red cells on the haemolytic capability of clinical isolates of Candida tropicalis , 2015, Journal of Biomedical Science.

[32]  P. Chong,et al.  Candida albicans isolates from a Malaysian hospital exhibit more potent phospholipase and haemolysin activities than non-albicans Candida isolates. , 2013, Tropical biomedicine.

[33]  M. Blanco-Blanco,et al.  Determination of biofilm production by Candida tropicalis isolated from hospitalized patients and its relation to cellular surface hydrophobicity, plastic adherence and filamentation ability , 2013, Yeast.

[34]  E. Ortega,et al.  Aminopeptidase N (CD13) Is Involved in Phagocytic Processes in Human Dendritic Cells and Macrophages , 2013, BioMed research international.

[35]  Jingyi Zhang,et al.  In vitro fluconazole susceptibility of 1,903 clinical isolates of Candida albicans and the identification of ERG11 mutations. , 2013, Microbial drug resistance.

[36]  J. Grimalt,et al.  Candida tropicalis Antifungal Cross-Resistance Is Related to Different Azole Target (Erg11p) Modifications , 2013, Antimicrobial Agents and Chemotherapy.

[37]  S. H. Alves,et al.  Determination of germ tube, phospholipase, and proteinase production by bloodstream isolates of Candida albicans. , 2013, Revista da Sociedade Brasileira de Medicina Tropical.

[38]  Beiqin Yu,et al.  Mechanisms of azole resistance in 52 clinical isolates of Candida tropicalis in China. , 2013, The Journal of antimicrobial chemotherapy.

[39]  D. Horn,et al.  Epidemiology and outcomes of candidemia in 3648 patients: data from the Prospective Antifungal Therapy (PATH Alliance®) registry, 2004-2008. , 2012, Diagnostic microbiology and infectious disease.

[40]  Rosário Oliveira,et al.  Insights into Candida tropicalis nosocomial infections and virulence factors , 2012, European Journal of Clinical Microbiology & Infectious Diseases.

[41]  M. Schaller,et al.  Candida albicans-Epithelial Interactions: Dissecting the Roles of Active Penetration, Induced Endocytosis and Host Factors on the Infection Process , 2012, PloS one.

[42]  C. D. de Koster,et al.  Hyphal induction in the human fungal pathogen Candida albicans reveals a characteristic wall protein profile. , 2011, Microbiology.

[43]  K. Ng,et al.  Proteinase, phospholipase, biofilm forming abilities and antifungal susceptibilities of Malaysian Candida isolates from blood cultures. , 2011, Medical mycology.

[44]  Rosário Oliveira,et al.  The role of secreted aspartyl proteinases in Candida tropicalis invasion and damage of oral mucosa. , 2011, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[45]  C. Tsang,et al.  HIV protease inhibitors differentially inhibit adhesion of Candida albicans to acrylic surfaces , 2010, Mycoses.

[46]  P. Le Pape,et al.  Screening for amino acid substitutions in the Candida albicans Erg11 protein of azole-susceptible and azole-resistant clinical isolates: new substitutions and a review of the literature. , 2010, Diagnostic microbiology and infectious disease.

[47]  N. Bannert,et al.  Cellular interactions of Candida albicans with human oral epithelial cells and enterocytes , 2010, Cellular microbiology.

[48]  David W Williams,et al.  Characterization of Candida parapsilosis infection of an in vitro reconstituted human oral epithelium. , 2009, European Journal of Oral Sciences.

[49]  C. D. de Koster,et al.  Of Novel Adhesin-like Wall Proteins : Differential Incorporation Candida Glabrata Supplemental Material , 2008 .

[50]  David F. Smith,et al.  Glycan microarray analysis of Candida glabrata adhesin ligand specificity , 2008, Molecular microbiology.

[51]  W. Leung,et al.  Phospholipase, proteinase and haemolytic activities of Candida albicans isolated from oral cavities of patients with type 2 diabetes mellitus. , 2007, Journal of medical microbiology.

[52]  C. Silveira,et al.  Enzymatic and hemolytic activities of Candida dubliniensis strains. , 2007, Revista do Instituto de Medicina Tropical de Sao Paulo.

[53]  M. Whiteway,et al.  Proteomic analysis of Candida albicans yeast and hyphal cell wall and associated proteins , 2006, Proteomics.

[54]  M. Schaller,et al.  Hydrolytic enzymes as virulence factors of Candida albicans , 2005, Mycoses.

[55]  D. Underhill,et al.  Dectin‐1 mediates macrophage recognition of Candida albicans yeast but not filaments , 2005, The EMBO journal.

[56]  A. Fluit,et al.  High levels of hydrolytic enzymes secreted by Candida albicans isolates involved in respiratory infections. , 2003, Journal of medical microbiology.

[57]  S. Challacombe,et al.  Candida albicans Secreted Aspartyl Proteinases in Virulence and Pathogenesis , 2003, Microbiology and Molecular Biology Reviews.

[58]  P. Chiani,et al.  Antiretroviral therapy with protease inhibitors has an early, immune reconstitution-independent beneficial effect on Candida virulence and oral candidiasis in human immunodeficiency virus-infected subjects. , 2002, The Journal of infectious diseases.

[59]  B. Wickes,et al.  Standardized Method for In Vitro Antifungal Susceptibility Testing of Candida albicansBiofilms , 2001, Antimicrobial Agents and Chemotherapy.

[60]  L. Samaranayake,et al.  Candida Species Exhibit Differential In Vitro Hemolytic Activities , 2001, Journal of Clinical Microbiology.

[61]  M. Dierich,et al.  HIV protease inhibitors attenuate adherence of Candida albicans to epithelial cells in vitro. , 2001, FEMS immunology and medical microbiology.

[62]  L. Samaranayake,et al.  Adhesion of Candida parapsilosis to epithelial and acrylic surfaces correlates with cell surface hydrophobicity , 2001, Mycoses.

[63]  M. Ghannoum Potential Role of Phospholipases in Virulence and Fungal Pathogenesis , 2000, Clinical Microbiology Reviews.

[64]  A. Telenti,et al.  HIV-Protease inhibitors reduce cell adherence of Candida albicans strains by inhibition of yeast secreted aspartic proteases. , 1999, The Journal of investigative dermatology.

[65]  E. Tacconelli,et al.  In vitro and in vivo anticandidal activity of human immunodeficiency virus protease inhibitors. , 1999, The Journal of infectious diseases.

[66]  E. Tacconelli,et al.  Role of protease inhibitors in preventing recurrent oral candidosis in patients with HIV infection: a prospective case-control study. , 1999, Journal of acquired immune deficiency syndromes.

[67]  A. Pachì,et al.  High Aspartyl Proteinase Production and Vaginitis in Human Immunodeficiency Virus-Infected Women , 1999, Journal of Clinical Microbiology.

[68]  A. Chakrabarti,et al.  In vitro proteinase production by Candida species , 1991, Mycopathologia.

[69]  J. Zajic,et al.  Factors governing adherence of Candida species to plastic surfaces , 1985, Infection and immunity.

[70]  M. Rosenberg Bacterial adherence to hydrocarbons: a useful technique for studying cell surface hydrophobicity , 1984 .

[71]  Shigeru Tsuchiya,et al.  Establishment and characterization of a human acute monocytic leukemia cell line (THP‐1) , 1980, International journal of cancer.

[72]  B. Kullberg,et al.  Invasive Candidiasis. , 2015, The New England journal of medicine.

[73]  C. Sachin,et al.  In vitro evaluation of proteinase, phospholipase and haemolysin activities of Candida species isolated from clinical specimens , 2012 .

[74]  M. Netea,et al.  An integrated model of the recognition of Candida albicans by the innate immune system , 2008, Nature Reviews Microbiology.