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Identification of Novel Hybrids Between Cryptococcus neoformans var. grubii VNI and Cryptococcus gattii VGII

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Abstract

Cryptococcus neoformans and Cryptococcus gattii are pathogenic yeasts causing meningoencephalitis in immunocompromised and immunocompetent hosts. The fungus is typically haploid, and sexual reproduction occurs normally between individuals with opposite mating types, α and a. C. neoformans var. grubii (serotype A) is comprised of molecular types VNI, VNII, and VNB, and C. neoformans var. neoformans (serotype D) contains the molecular type VNIV. Additionally, diploid or aneuploid AD hybrids (VNIII) have been reported. C. gattii contains the molecular types VGI, VGII, VGIII, and VGIV, which encompass both serotypes B and C. To identify possible hybrid strains, URA5-RFLP analysis was performed on 350 globally obtained clinical, environmental, and veterinary isolates. Four clinical isolates from cerebrospinal fluid showed combination patterns of C. neoformans var. grubii and C. gattii: Brazil (n = 2), Colombia (n = 1), and India (n = 1). These strains were monokaryotic and diploid or aneuploid. M13 PCR fingerprinting showed that they contained fragments of both proposed parental groups. Luminex IGS genotyping identified these isolates as hybrids with two different molecular type combinations: three VNI/VGII and one VNI/VGI. Blue color development on CGB agar was delayed in three isolates and absent in one. C. gattii-specific PCR confirmed the presence of C. gattii in the hybrids. CAP59 allele-specific PCR revealed that all the hybrids contained both serotype A and B alleles. Determination of mating-type allelic patterns by PCR revealed that the isolates were αA aB. This is the first study discovering novel natural hybrids between C. neoformans molecular type VNI and C. gattii molecular type VGII.

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Acknowledgments

The authors thank Mary Mattews for providing the strain from India; Maggie Wang and Stephen Schibeci for technical assistance on flow cytometry analysis; and Fottes Al. Panetsos and Hariklia Moshoyiannis for technical assistance on Luminex analysis. This work was supported by a University of Sydney Postgraduate Award and a CIDM Public health scholarship to MAm. The work of the Sydney Institute for Emerging Infections and Biosecurity is supported by a grant from the Sydney Medical School Foundation. TCS is a Sydney Medical School Foundation Fellow. The Bodosakis Foundation and the Research fund SARG 70/3/6915 National and Kapodistrian University of Athens partially funded this work. Flow cytometry was performed in the Flow Cytometry Core Facility that is supported by Westmead Millennium Institute, National Health and Medical Research Council and Cancer Institute New South Wales, Australia.

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Authors and Affiliations

  1. Molecular Mycology Research Laboratory, Westmead, Australia

    Mojgan Aminnejad

  2. Centre for Infectious Diseases and Microbiology, Sydney Medical School-Westmead, Westmead Millennium Institute, Sydney Emerging Infections and Biosecurity Institute, University of Sydney, Sydney, NSW, Australia

    Mojgan Aminnejad, Tania C. Sorrell & Wieland Meyer

  3. Rosenstiel School of Marine and Atmospheric Sciences, University of Miami, Miami, USA

    Mara Diaz

  4. Mycology Laboratory, Department of Microbiology, Medical School, National and Kapodistrian University of Athens, Athens, Greece

    Michael Arabatzis & Aristea Velegraki

  5. Grupo de Microbiología, Instituto Nacional de Salud, Bogotá, Colombia

    Elizabeth Castañeda

  6. Mycology Laboratory, Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil

    Marcia Lazera

  7. Department of Microbiology and Clinical Infectious Diseases, St. Vincent’s Hospital, Sydney, NSW, Australia

    Deborah Marriott

  8. Molecular Mycology Research Laboratory, CIDM, ICPMR, Level 3, Room 3114A, Westmead Hospital, Darcy Road, Westmead, NSW, 2145, Australia

    Wieland Meyer

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  1. Mojgan Aminnejad
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Aminnejad, M., Diaz, M., Arabatzis, M. et al. Identification of Novel Hybrids Between Cryptococcus neoformans var. grubii VNI and Cryptococcus gattii VGII. Mycopathologia 173, 337–346 (2012). https://doi.org/10.1007/s11046-011-9491-x

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