Identification of Virulence Factors in Isolates of Candida haemulonii, Candida albicans and Clavispora lusitaniae with Low Susceptibility and Resistance to Fluconazole and Amphotericin B
Abstract
:1. Introduction
2. Materials and Methods
2.1. Microorganisms and Growth Conditions
2.2. Antifungal Activity of Fluconazole and Amphotericin B
2.3. Hydrophobicity Assay
2.4. Adherence on Plastic Surface by Crystal Violet Assay
2.5. Biofilm Formation
2.6. Lipase Activity Assay
2.7. Sensitivity to Osmotic Stress in Sodium Chloride
2.8. Infection in Galleria Mellonella Model
2.9. Statistical Analysis
3. Results
3.1. Susceptibility to Fluconazole and Amphotericin B
3.2. Hydrophobicity Assay and Adherence on Plastic Surface
3.3. Biofilm Formation
3.4. Lipase Activity and Osmotic Stress
3.5. Galleria Mellonella Survival Assay
4. Discussion
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Temkin, E.; Adler, A.; Lerner, A.; Carmeli, Y. Carbapenem-resistant Enterobacteriaceae: Biology, epidemiology, and management. Ann. N. Y. Acad. Sci. 2014, 1323, 22–42. [Google Scholar] [CrossRef] [PubMed]
- Humphreys, G.; Fleck, F. United Nations meeting on antimicrobial resistance. Bull. World Health Organ. 2016, 94, 638–639. [Google Scholar]
- Stevenson, E.M.; Gaze, W.H.; Gow, N.A.R.; Hart, A. Antifungal Exposure and Resistance Development: Defining Minimal Selective Antifungal Concentrations and Testing Methodologies. Front. Fungal Biol. 2022, 3, 918717. [Google Scholar] [CrossRef] [PubMed]
- Hossain, C.M.; Ryan, L.K.; Gera, M.; Choudhuri, S.; Lyle, N.; Ali, K.A.; Diamond, G. Antifungal and drug resistance. Encyclopedia 2022, 2, 1722–1737. [Google Scholar] [CrossRef]
- Ramos, L.S.; Figueiredo-Carvalho, M.H.G.; Silva, L.N.; Siqueira, N.L.; Lima, J.C.; Oliveira, S.S.; Almeida-Paes, R.; Zancopé-Oliveira, R.M.; Azevedo, F.S.; Ferreira, A.L.; et al. The Threat Called Candida haemulonii Species Complex in Rio de Janeiro State, Brazil: Focus on Antifungal Resistance and Virulence Attributes. J. Fungi 2022, 8, 574. [Google Scholar] [CrossRef] [PubMed]
- Cendejas-Bueno, E.; Kolecka, A.; Alastruey-Izquierdo, A.; Theelen, B.; Groenewald, M.; Kostrzewa, M.; Cuenca-Estrella, M.; Gomez-Lopez, A.; Boekhout, T. Reclassification of the Candida haemulonii complex as Candida haemulonii (C. haemulonii group I), C. duobushaemulonii sp. nov. (C. haemulonii group II), and C. haemulonii var. vulnera var. nov.: Three multiresistant human pathogenic yeasts. J. Clin. Microbiol. 2012, 50, 3641–3651. [Google Scholar] [PubMed]
- Ben-Ami, R.B.; Berman, J.; Novikov, A. Multi-drug resistant Candida haemulonii and C. auris, Tel Aviv, Israel. Emerg. Infect. Dis. 2017, 23, 195–203. [Google Scholar] [CrossRef]
- Lima, S.L.; Rossato, L.; Salles de Azevedo Melo, A. Evaluation of the potential virulence of Candida haemulonii species complex and Candida auris isolates in Caenorhabditis elegans as an in vivo model and correlation to their biofilm production capacity. Microbial Pathogenesis. 2020, 148, 104461. [Google Scholar] [CrossRef]
- Silva, L.N.; Oliveira, S.S.; Magalhães, L.B.; Andrade Neto, V.V.; Torres-Santos, E.C.; Carvalho, M.D.; Pereira, M.D.; Branquinha, M.H.; Santos, A.L. Unmasking the Amphotericin B Resistance Mechanisms in Candida haemulonii Species Complex. ACS Infect. Dis. 2020, 6, 1273–1282. [Google Scholar] [CrossRef]
- Deng, Y.; Li, S.; Bing, J.; Liao, W.; Tao, L. Phenotypic Switching and Filamentation in Candida haemulonii, an Emerging Opportunistic Pathogen of Humans. Microbiol. Spectrum 2021, 9, e00779-21. [Google Scholar] [CrossRef]
- Pappagianis, D.; Collins, M.S.; Hector, R.; Remington, J. Development of resistance to amphotericin B in Candida lusitaniae infecting a human. Antimicrob. Agents Chemother. 1979, 16, 123–126. [Google Scholar] [CrossRef] [PubMed]
- Fakhim, H.; Vaezi, A.; Dannaoui, E.; Chowdhary, A.; Nasiry, D.; Faeli, L.; Meis, J.F.; Badali, H. Comparative virulence of Candida auris with Candida haemulonii, Candida glabrata and Candida albicans in a murine model. Mycoses 2018, 61, 377–382. [Google Scholar] [CrossRef] [PubMed]
- Mendoza-Reyes, D.; Gómez-Gaviria, M.; Mora-Montes, H. Candida lusitaniae: Biology, Pathogenicity, Virulence Factors, Diagnosis, and Treatment. Infect. Drug Resist. 2022, 15, 5121–5135. [Google Scholar] [CrossRef] [PubMed]
- Jung, D.S.; Farmakiotis, D.; Jiang, Y.; Tarrand, J.J.; Kontoyiannis, D.P. Uncommon Candida species fungemia among cancer patients, Houston, Texas, USA. Emerg. Infect. Dis. 2015, 21, 1942–1950. [Google Scholar] [CrossRef]
- Obisesan, O.J.; Olowe, O.A.; Taiwo, S.S. Phenotypic detection of genitourinary candidiasis among sexually transmitted disease clinic attendees in Ladoke Akintola University Teaching Hospital, Osogbo, Nigeria. J. Environ. Public. Health. 2015, 2015, 401340. [Google Scholar] [CrossRef] [PubMed]
- Angiolella, L.; Stringaro, A.R.; De Bernardis, F.; Posteraro, B.; Bonito, M.; Toccacieli, L.; Torosantucci, A.; Colone, M.; Sanguinetti, M.; Cassone, A.; et al. Increase of Virulence and Its Phenotypic Traits in Drug-Resistant Strains of Candida albicans. Antimicrob. Agents Chemother. 2008, 52, 927–936. [Google Scholar] [CrossRef] [PubMed]
- Rajendran, R.; Sherry, L.; Nile, C.J.; Sherriff, A.; Johnson, E.M.; Hanson, M.F.; Williams, C.; Munro, C.A.; Jones, B.J.; Ramage, G. Biofilm formation is a risk factor for mortality in patients with Candida albicans bloodstream infection—Scotland, 2012–2013. Clin. Microbiol. Infect. 2016, 22, 87–93. [Google Scholar] [CrossRef]
- Zhou, Y.; Li, G.; Dong, J.; ** of Candida haemulonii, Candida pseudohaemulonii, and a proposed new species (Candida auris) isolates from Korea. Med. Mycol. 2011, 49, 98–102. [Google Scholar] [CrossRef]
- Santos, A.L.S.; Mello, T.P.; Ramos, L.S.; Branquinha, M.H. Biofilm: A robust and efficient barrier to antifungal chemotherapy. J. Antimicrob. 2015, 1, e101. [Google Scholar] [CrossRef]
- Mukherjee, P.K.; Chandra, J. Candida biofilms: Development, architecture, and resistance. Microbiol. Spectr. 2015, 3, 115–134. [Google Scholar]
- Kean, R.; Delaney, C.; Sherry, L.; Borman, A.; Johnson, E.M.; Richardson, M.D.; Rautemaa-Richardson, R.; Williams, C.; Ramage, G. Transcriptome assembly and profiling of Candida auris reveals novel insights into biofilm-mediated resistance. mSphere 2018, 3, e00334-18. [Google Scholar] [CrossRef] [PubMed]
- Park, M.; Do, E.; Jung, W.H. Lipolytic enzymes involved in the virulence of human pathogenic fungi. Mycobiology 2013, 41, 67–72. [Google Scholar] [CrossRef]
- Brown, A.J.P.; Brown, G.D.; Netea, M.G.; Gow, N.A.R. Metabolism impacts upon Candida immunogenicity and pathogenicity at multiple levels. Trends Microbiol. 2014, 22, 614–622. [Google Scholar] [CrossRef]
- Brown, A.J.; Budge, S.; Kaloriti, D.; Tillmann, A.; Jacobsen, M.D.; Yin, Z.; Ene, I.V.; Bohovych, I.; Sandai, D.; Kastora, S.; et al. Stress adaptation in a pathogenic fungus. J. Exp. Biol. 2013, 217, 144–155. [Google Scholar] [CrossRef]
- Brown, A.J.P.; Haynes, K.; Gow, N.A.R.; Quinn, J. Stress Responses in Candida. In Candida and Candidiasis; Calderone, R.A., Clancy, C.J., Eds.; ASM Press: Washington, DC, USA, 2012; pp. 225–242. [Google Scholar] [CrossRef]
Fluconazole | Amphotericin B | |
---|---|---|
Strains | MIC (µg/mL) | MIC (µg/mL) |
IMR-M-L 1462 C. albicans | 8 | 0.5 |
IMR-M-L 1463 C. albicans | 4 | 0.5 |
IMR-M-L 1464 C. albicans | 4 | 0.5 |
ATCC 24433 C. albicans | 0.25 | 0.5 |
IMR-M-L 301 C. lusitaniae | 0.5 | 1 |
IMR-M-L 522 C. lusitaniae | 2 | 2 |
IMR-M-L 1112 C. lusitaniae | 2 | 2 |
IMR-M-L 1384 C. lusitaniae | 4 | >16 |
IMR-M-L 785 C. haemulonii | 32 | 16 |
IMR-M-L 1293 C. haemulonii | >64 | 16 |
IMR-M-L 1375 C. haemulonii | 4 | 2 |
MIC range | 0.25 > 64 | 0.5 > 16 |
Strains | Lz Index | Activity | Mean LZ ± SD |
---|---|---|---|
IMR-M-L 1462 C. albicans | 0.42 | High | 0.44 ± 0.06 |
IMR-M-L 1463 C. albicans | 0.53 | High | |
IMR-M-L 1464 C. albicans | 0.37 | High | |
ATCC 24433 C. albicans | 0.46 | High | |
IMR-M-L 301 C. lusitaniae | 1.00 | None | 0.92 ± 0.13 **** |
IMR-M-L 522 C. lusitaniae | 1.00 | None | |
IMR-M-L 1112 C. lusitaniae | 1.00 | None | |
IMR-M-L 1384 C. lusitaniae | 0.70 | Moderate | |
IMR-M-L 785 C. haemulonii | 1.00 | High | 0.73 ± 0.25 **** |
IMR-M-L 1293 C. haemulonii | 0.70 | Moderate | |
IMR-M-L 1375 C. haemulonii | 0.49 | High |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Angiolella, L.; Rojas, F.; Giammarino, A.; Bellucci, N.; Giusiano, G. Identification of Virulence Factors in Isolates of Candida haemulonii, Candida albicans and Clavispora lusitaniae with Low Susceptibility and Resistance to Fluconazole and Amphotericin B. Microorganisms 2024, 12, 212. https://doi.org/10.3390/microorganisms12010212
Angiolella L, Rojas F, Giammarino A, Bellucci N, Giusiano G. Identification of Virulence Factors in Isolates of Candida haemulonii, Candida albicans and Clavispora lusitaniae with Low Susceptibility and Resistance to Fluconazole and Amphotericin B. Microorganisms. 2024; 12(1):212. https://doi.org/10.3390/microorganisms12010212
Chicago/Turabian StyleAngiolella, Letizia, Florencia Rojas, Andrea Giammarino, Nicolò Bellucci, and Gustavo Giusiano. 2024. "Identification of Virulence Factors in Isolates of Candida haemulonii, Candida albicans and Clavispora lusitaniae with Low Susceptibility and Resistance to Fluconazole and Amphotericin B" Microorganisms 12, no. 1: 212. https://doi.org/10.3390/microorganisms12010212