Cryptococcus Genetic Diversity and Mixed Infections in Ivorian HIV Patients: A Follow Up Study

Overview

Journal PLoS Negl Trop Dis

Specialties Infectious Diseases
Tropical Medicine

Date 2019 Nov 19

PMID 31738768

Citations 13

Authors

Fulgence Kondo Kassi

Pascal Drakulovski

Virginie Bellet

Frederic Roger

Amelie Chabrol

Donika Krasteva

Adama Doumbia

Roland Landman

Aka Kakou

Jacques Reynes

Eric Delaporte

Herve Eby Ignace Menan

Sebastien Bertout

Affiliations

Soon will be listed here.

Abstract

Genetic diversity analyses were performed by sero-genotyping and multi-locus sequence typing on 252 cryptococcal isolates from 13 HIV-positive Ivorian patients followed-up for cryptococcal meningitis. Antifungal susceptibility analyses were performed according to the CLSI M27A3 method. The majority (67.8%) of the isolates belonged to the Cryptococcus neoformans (serotype A) species complex, with 93% being VNI and 7% being VNII. Cryptococcus deuterogattii VGII (serotype B) represented 16.7% of the strains, while C. neoformans/C. deneoformans VNIII (serotype AD) hybrids accounted for 15.1% of the strains. One strain (0.4%) was not identifiable. Nine different sequence types (STs 5, 6, 23, 40, 93, 207, 311, and a new ST; 555) were identified in the C. neoformans population, while the C. deuterogattii population comprised the single ST 173. The distribution of the strains showed that, while the majority of patients (9/13) harboured a single sequence type, 4 patients showed mixed infections. These patients experienced up to 4 shifts in strain content either at the species and/or ST level during their follow-up. This evolution of diversity over time led to the co-existence of up to 3 different Cryptococcus species and 4 different ST within the same individual during the course of infection. Susceptibility testing showed that all strains were susceptible to amphotericin B while 3.6% of them had a none-wild type phenotype to 5-flucytosine. Concerning fluconazole, 2.9% of C.neoformans serotype A strains and 2.4% of C. deuterogattii had also respectively a none-wild type phenotype to this molecule. All C. neoformans x C. deneoformans serotype AD hybrids had however a wild type phenotype to fluconazole. The present study showed that mixed infections exist and could be of particular importance for care outcomes. Indeed, (i) the different Cryptococcus species are known to exhibit different virulence and different susceptibility patterns to antifungal drugs and (ii) the strains genetic diversity within the samples may influence the susceptibility to antifungal treatment.

Citing Articles

Antifungal Resistance in Cryptococcal Infections.

Melhem M, Leite Junior D, Takahashi J, Macioni M, Oliveira L, de Araujo L Pathogens. 2024; 13(2).

PMID: 38392866 PMC: 10891860. DOI: 10.3390/pathogens13020128.

Genetic diversity and microevolution in clinical Cryptococcus isolates from Cameroon.

Sephton-Clark P, Temfack E, Tenor J, Toffaletti D, Loyse A, Molloy S Med Mycol. 2023; 61(12).

PMID: 37952096 PMC: 10709296. DOI: 10.1093/mmy/myad116.

Genotyping Analysis of and Correlation with Virulence Factors and Antifungal Susceptibility by the Clinical and Laboratory Standards Institute and the European Committee on Antifungal Susceptibility Testing Methods.

Andrade-Silva L, Vilas-Boas A, Ferreira-Paim K, Andrade-Silva J, Santos D, Ferreira T J Fungi (Basel). 2023; 9(9).

PMID: 37754997 PMC: 10532325. DOI: 10.3390/jof9090889.

Fluconazole Resistance and Virulence in In Vitro Induced-Fluconazole Resistant Strains and in Clinical Fluconazole Resistant Strain of .

Bertout S, Laroche L, Roger F, Krasteva D, Drakulovski P, Bellet V Pathogens. 2023; 12(6).

PMID: 37375448 PMC: 10304499. DOI: 10.3390/pathogens12060758.

Exposure of to Seven Commonly Used Agricultural Azole Fungicides Induces Resistance to Fluconazole as Well as Cross-Resistance to Voriconazole, Posaconazole, Itraconazole and Isavuconazole.

Drakulovski P, Krasteva D, Bellet V, Randazzo S, Roger F, Pottier C Pathogens. 2023; 12(5).

PMID: 37242332 PMC: 10224184. DOI: 10.3390/pathogens12050662.

References

Murphy R, Hatlen T, Moosa M . High-Dose Fluconazole Consolidation Therapy for Cryptococcal Meningitis in Sub-Saharan Africa: Much to Gain, Little to Lose. AIDS Res Hum Retroviruses. 2018; 34(5):399-403. DOI: 10.1089/AID.2017.0240. View

Kassi F, Bellet V, Drakulovski P, Krasteva D, Roger F, Valerie B . Comparative typing analyses of clinical and environmental strains of the Cryptococcus neoformans/Cryptococcus gattii species complex from Ivory Coast. J Med Microbiol. 2017; 67(1):87-96. DOI: 10.1099/jmm.0.000654. View

Klein K, Hall L, Deml S, Rysavy J, Wohlfiel S, Wengenack N . Identification of Cryptococcus gattii by use of L-canavanine glycine bromothymol blue medium and DNA sequencing. J Clin Microbiol. 2009; 47(11):3669-72. PMC: 2772631. DOI: 10.1128/JCM.01072-09. View

Danesi P, Firacative C, Cogliati M, Otranto D, Capelli G, Meyer W . Multilocus sequence typing (MLST) and M13 PCR fingerprinting revealed heterogeneity amongst Cryptococcus species obtained from Italian veterinary isolates. FEMS Yeast Res. 2014; 14(6):897-909. DOI: 10.1111/1567-1364.12178. View

Martins M, Pappalardo M, Melhem M, Pereira-Chioccola V . Molecular diversity of serial Cryptococcus neoformans isolates from AIDS patients in the city of São Paulo, Brazil. Mem Inst Oswaldo Cruz. 2007; 102(7):777-84. DOI: 10.1590/s0074-02762007000700001. View

Litvintseva A, Thakur R, Vilgalys R, Mitchell T . Multilocus sequence typing reveals three genetic subpopulations of Cryptococcus neoformans var. grubii (serotype A), including a unique population in Botswana. Genetics. 2005; 172(4):2223-38. PMC: 1456387. DOI: 10.1534/genetics.105.046672. View

Hagen F, Illnait-Zaragozi M, Meis J, Chew W, Curfs-Breuker I, Mouton J . Extensive genetic diversity within the Dutch clinical Cryptococcus neoformans population. J Clin Microbiol. 2012; 50(6):1918-26. PMC: 3372159. DOI: 10.1128/JCM.06750-11. View

Rozas J, Ferrer-Mata A, Sanchez-DelBarrio J, Guirao-Rico S, Librado P, Ramos-Onsins S . DnaSP 6: DNA Sequence Polymorphism Analysis of Large Data Sets. Mol Biol Evol. 2017; 34(12):3299-3302. DOI: 10.1093/molbev/msx248. View

Vanhove M, Beale M, Rhodes J, Chanda D, Lakhi S, Kwenda G . Genomic epidemiology of Cryptococcus yeasts identifies adaptation to environmental niches underpinning infection across an African HIV/AIDS cohort. Mol Ecol. 2016; 26(7):1991-2005. PMC: 5412878. DOI: 10.1111/mec.13891. View

10.

Sanchini A, McCormick Smith I, Sedlacek L, Schwarz R, Tintelnot K, Rickerts V . Molecular typing of clinical Cryptococcus neoformans isolates collected in Germany from 2004 to 2010. Med Microbiol Immunol. 2014; 203(5):333-40. DOI: 10.1007/s00430-014-0341-6. View

11.

Gutch R, Nawange S, Singh S, Yadu R, Tiwari A, Gumasta R . Antifungal susceptibility of clinical and environmental Cryptococcus neoformans and Cryptococcus gattii isolates in Jabalpur, a city of Madhya Pradesh in Central India. Braz J Microbiol. 2015; 46(4):1125-33. PMC: 4704646. DOI: 10.1590/S1517-838246420140564. View

12.

Souto A, Bonfietti L, Ferreira-Paim K, Trilles L, Martins M, Ribeiro-Alves M . Population Genetic Analysis Reveals a High Genetic Diversity in the Brazilian Cryptococcus gattii VGII Population and Shifts the Global Origin from the Amazon Rainforest to the Semi-arid Desert in the Northeast of Brazil. PLoS Negl Trop Dis. 2016; 10(8):e0004885. PMC: 4986980. DOI: 10.1371/journal.pntd.0004885. View

13.

Desnos-Ollivier M, Patel S, Spaulding A, Charlier C, Garcia-Hermoso D, Nielsen K . Mixed infections and In Vivo evolution in the human fungal pathogen Cryptococcus neoformans. mBio. 2010; 1(1). PMC: 2912664. DOI: 10.1128/mBio.00091-10. View

14.

Fan X, Xiao M, Chen S, Kong F, Dou H, Wang H . Predominance of Cryptococcus neoformans var. grubii multilocus sequence type 5 and emergence of isolates with non-wild-type minimum inhibitory concentrations to fluconazole: a multi-centre study in China. Clin Microbiol Infect. 2016; 22(10):887.e1-887.e9. DOI: 10.1016/j.cmi.2016.07.008. View

15.

Grosset M, Desnos-Ollivier M, Godet C, Kauffmann-Lacroix C, Cazenave-Roblot F . Recurrent episodes of Candidemia due to and with acquired echinocandin resistance. Med Mycol Case Rep. 2016; 14:20-23. PMC: 5155041. DOI: 10.1016/j.mmcr.2016.12.004. View

16.

Longley N, Muzoora C, Taseera K, Mwesigye J, Rwebembera J, Chakera A . Dose response effect of high-dose fluconazole for HIV-associated cryptococcal meningitis in southwestern Uganda. Clin Infect Dis. 2008; 47(12):1556-61. DOI: 10.1086/593194. View

17.

Sionov E, Chang Y, Garraffo H, Kwon-Chung K . Heteroresistance to fluconazole in Cryptococcus neoformans is intrinsic and associated with virulence. Antimicrob Agents Chemother. 2009; 53(7):2804-15. PMC: 2704677. DOI: 10.1128/AAC.00295-09. View

18.

Assogba K, Belo M, Wateba M, Gnonlonfoun D, Ossou-Nguiet P, Tsanga B . Neuromeningeal cryptococcosis in sub-Saharan Africa: Killer disease with sparse data. J Neurosci Rural Pract. 2015; 6(2):221-4. PMC: 4387815. DOI: 10.4103/0976-3147.153231. View

19.

Milefchik E, Leal M, Haubrich R, Bozzette S, Tilles J, Leedom J . Fluconazole alone or combined with flucytosine for the treatment of AIDS-associated cryptococcal meningitis. Med Mycol. 2008; 46(4):393-5. DOI: 10.1080/13693780701851695. View

20.

Meyer W, Aanensen D, Boekhout T, Cogliati M, Diaz M, Esposto M . Consensus multi-locus sequence typing scheme for Cryptococcus neoformans and Cryptococcus gattii. Med Mycol. 2009; 47(6):561-70. PMC: 2884100. DOI: 10.1080/13693780902953886. View