Identification and Characterization of VNI/VNII and Novel VNII/VNIV Hybrids and Impact of Hybridization on Virulence and Antifungal Susceptibility Within the C. neoformans/C. gattii Species Complex
Cryptococcus neoformans and C. gattii are pathogenic basidiomycetous yeasts and the commonest cause of fungal infection of the central nervous system. Cryptococci are typically haploid but several inter-species, inter-varietal and intra-varietal hybrids have been reported. It has a bipolar mating sy...
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| creator | Aminnejad, Mojgan Cogliati, Massimo Duan, Shuyao Arabatzis, Michael Tintelnot, Kathrin Castañeda, Elizabeth Lazéra, Marcia Velegraki, Aristea Ellis, David Sorrell, Tania C Meyer, Wieland |
| description | Cryptococcus neoformans and C. gattii are pathogenic basidiomycetous yeasts and the commonest cause of fungal infection of the central nervous system. Cryptococci are typically haploid but several inter-species, inter-varietal and intra-varietal hybrids have been reported. It has a bipolar mating system with sexual reproduction occurring normally between two individuals with opposite mating types, α and a. This study set out to characterize hybrid isolates within the C. neoformans/C. gattii species complex: seven unisexual mating intra-varietal VNI/VNII (αAAα) and six novel inter-varietal VNII/VNIV (aADα). The URA5-RFLP pattern for VNII/VNIV (aADα) differs from the VNIII (αADa) hybrids. Analysis of the allelic patterns of selected genes for AD hybrids showed 79% or more heterozygosis for the studied loci except for CBS132 (VNIII), which showed 50% of heterozygosity. MALDI-TOF MS was applied to hybrids belonging to different sero/mating type allelic patterns. All hybrid isolates were identified as belonging to the same hybrid group with identification scores ranging between 2.101 to 2.634. All hybrids were virulent when tested in the Galleria mellonella (wax moth) model, except for VNII/VNIV (aADα) hybrids. VNI/VGII hybrids were the most virulent hybrids. Hybrids recovered from larvae manifested a significant increase in capsule and total cell size and produced a low proportion (5-10%) of giant cells compared with the haploid control strains. All strains expressed the major virulence factors-capsule, melanin and phospholipase B-and grew well at 37°C. The minimal inhibitory concentration of nine drugs was measured by micro-broth dilution and compared with published data on haploid strains. MICs were similar amongst hybrids and haploid parental strains. This is the first study reporting natural same sex αAAα intra-varietal VNI/VNII hybrids and aADα inter-varietal VNII/VNIV hybrids. |
| doi_str_mv | 10.1371/journal.pone.0163955 |
| format | Article |
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Cryptococci are typically haploid but several inter-species, inter-varietal and intra-varietal hybrids have been reported. It has a bipolar mating system with sexual reproduction occurring normally between two individuals with opposite mating types, α and a. This study set out to characterize hybrid isolates within the C. neoformans/C. gattii species complex: seven unisexual mating intra-varietal VNI/VNII (αAAα) and six novel inter-varietal VNII/VNIV (aADα). The URA5-RFLP pattern for VNII/VNIV (aADα) differs from the VNIII (αADa) hybrids. Analysis of the allelic patterns of selected genes for AD hybrids showed 79% or more heterozygosis for the studied loci except for CBS132 (VNIII), which showed 50% of heterozygosity. MALDI-TOF MS was applied to hybrids belonging to different sero/mating type allelic patterns. All hybrid isolates were identified as belonging to the same hybrid group with identification scores ranging between 2.101 to 2.634. All hybrids were virulent when tested in the Galleria mellonella (wax moth) model, except for VNII/VNIV (aADα) hybrids. VNI/VGII hybrids were the most virulent hybrids. Hybrids recovered from larvae manifested a significant increase in capsule and total cell size and produced a low proportion (5-10%) of giant cells compared with the haploid control strains. All strains expressed the major virulence factors-capsule, melanin and phospholipase B-and grew well at 37°C. The minimal inhibitory concentration of nine drugs was measured by micro-broth dilution and compared with published data on haploid strains. MICs were similar amongst hybrids and haploid parental strains. This is the first study reporting natural same sex αAAα intra-varietal VNI/VNII hybrids and aADα inter-varietal VNII/VNIV hybrids.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0163955</identifier><identifier>PMID: 27764108</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Antifungal agents ; Antifungal Agents - pharmacology ; Biology and Life Sciences ; Cell size ; Central nervous system ; Cryptococcus ; Cryptococcus gattii ; Cryptococcus gattii - genetics ; Cryptococcus gattii - physiology ; Cryptococcus neoformans ; Cryptococcus neoformans - genetics ; Cryptococcus neoformans - physiology ; Dilution ; Fungal infections ; Fungal Proteins - genetics ; Fungal Proteins - metabolism ; Fungicides ; Galleria mellonella ; Genes, Mating Type, Fungal - genetics ; Giant cells ; Haploidy ; Health aspects ; Heterozygosity ; HIV ; Human immunodeficiency virus ; Hybridization ; Hybridization, Genetic - genetics ; Hybrids ; Infections ; Infectious diseases ; Laboratories ; Larvae ; Lysophospholipase - metabolism ; Mating ; Mating types ; Medical research ; Medical schools ; Medicine and Health Sciences ; Melanin ; Melanins - metabolism ; Microbial Sensitivity Tests ; Morphology ; Moths ; Multiplex Polymerase Chain Reaction ; Mycological Typing Techniques ; Mycoses ; Pathogens ; Phospholipase ; Physical Sciences ; Reproduction (biology) ; Research and Analysis Methods ; Serogroup ; Sexual reproduction ; Species ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Virulence ; Virulence (Microbiology) ; Virulence - drug effects ; Virulence - genetics ; Virulence factors ; Virulence Factors - analysis ; Virulence Factors - genetics ; Virulence Factors - metabolism ; Yeasts</subject><ispartof>PloS one, 2016-10, Vol.11 (10), p.e0163955-e0163955</ispartof><rights>COPYRIGHT 2016 Public Library of Science</rights><rights>2016 Aminnejad et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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Cryptococci are typically haploid but several inter-species, inter-varietal and intra-varietal hybrids have been reported. It has a bipolar mating system with sexual reproduction occurring normally between two individuals with opposite mating types, α and a. This study set out to characterize hybrid isolates within the C. neoformans/C. gattii species complex: seven unisexual mating intra-varietal VNI/VNII (αAAα) and six novel inter-varietal VNII/VNIV (aADα). The URA5-RFLP pattern for VNII/VNIV (aADα) differs from the VNIII (αADa) hybrids. Analysis of the allelic patterns of selected genes for AD hybrids showed 79% or more heterozygosis for the studied loci except for CBS132 (VNIII), which showed 50% of heterozygosity. MALDI-TOF MS was applied to hybrids belonging to different sero/mating type allelic patterns. All hybrid isolates were identified as belonging to the same hybrid group with identification scores ranging between 2.101 to 2.634. All hybrids were virulent when tested in the Galleria mellonella (wax moth) model, except for VNII/VNIV (aADα) hybrids. VNI/VGII hybrids were the most virulent hybrids. Hybrids recovered from larvae manifested a significant increase in capsule and total cell size and produced a low proportion (5-10%) of giant cells compared with the haploid control strains. All strains expressed the major virulence factors-capsule, melanin and phospholipase B-and grew well at 37°C. The minimal inhibitory concentration of nine drugs was measured by micro-broth dilution and compared with published data on haploid strains. MICs were similar amongst hybrids and haploid parental strains. 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metabolism</topic><topic>Mating</topic><topic>Mating types</topic><topic>Medical research</topic><topic>Medical schools</topic><topic>Medicine and Health Sciences</topic><topic>Melanin</topic><topic>Melanins - metabolism</topic><topic>Microbial Sensitivity Tests</topic><topic>Morphology</topic><topic>Moths</topic><topic>Multiplex Polymerase Chain Reaction</topic><topic>Mycological Typing Techniques</topic><topic>Mycoses</topic><topic>Pathogens</topic><topic>Phospholipase</topic><topic>Physical Sciences</topic><topic>Reproduction (biology)</topic><topic>Research and Analysis Methods</topic><topic>Serogroup</topic><topic>Sexual reproduction</topic><topic>Species</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</topic><topic>Virulence</topic><topic>Virulence (Microbiology)</topic><topic>Virulence - drug effects</topic><topic>Virulence - genetics</topic><topic>Virulence factors</topic><topic>Virulence Factors - analysis</topic><topic>Virulence Factors - genetics</topic><topic>Virulence Factors - 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Academic</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Materials Science Collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Aminnejad, Mojgan</au><au>Cogliati, Massimo</au><au>Duan, Shuyao</au><au>Arabatzis, Michael</au><au>Tintelnot, Kathrin</au><au>Castañeda, Elizabeth</au><au>Lazéra, Marcia</au><au>Velegraki, Aristea</au><au>Ellis, David</au><au>Sorrell, Tania C</au><au>Meyer, Wieland</au><au>Nielsen, Kirsten</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification and Characterization of VNI/VNII and Novel VNII/VNIV Hybrids and Impact of Hybridization on Virulence and Antifungal Susceptibility Within the C. neoformans/C. gattii Species Complex</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2016-10-20</date><risdate>2016</risdate><volume>11</volume><issue>10</issue><spage>e0163955</spage><epage>e0163955</epage><pages>e0163955-e0163955</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Cryptococcus neoformans and C. gattii are pathogenic basidiomycetous yeasts and the commonest cause of fungal infection of the central nervous system. Cryptococci are typically haploid but several inter-species, inter-varietal and intra-varietal hybrids have been reported. It has a bipolar mating system with sexual reproduction occurring normally between two individuals with opposite mating types, α and a. This study set out to characterize hybrid isolates within the C. neoformans/C. gattii species complex: seven unisexual mating intra-varietal VNI/VNII (αAAα) and six novel inter-varietal VNII/VNIV (aADα). The URA5-RFLP pattern for VNII/VNIV (aADα) differs from the VNIII (αADa) hybrids. Analysis of the allelic patterns of selected genes for AD hybrids showed 79% or more heterozygosis for the studied loci except for CBS132 (VNIII), which showed 50% of heterozygosity. MALDI-TOF MS was applied to hybrids belonging to different sero/mating type allelic patterns. All hybrid isolates were identified as belonging to the same hybrid group with identification scores ranging between 2.101 to 2.634. All hybrids were virulent when tested in the Galleria mellonella (wax moth) model, except for VNII/VNIV (aADα) hybrids. VNI/VGII hybrids were the most virulent hybrids. Hybrids recovered from larvae manifested a significant increase in capsule and total cell size and produced a low proportion (5-10%) of giant cells compared with the haploid control strains. All strains expressed the major virulence factors-capsule, melanin and phospholipase B-and grew well at 37°C. The minimal inhibitory concentration of nine drugs was measured by micro-broth dilution and compared with published data on haploid strains. MICs were similar amongst hybrids and haploid parental strains. This is the first study reporting natural same sex αAAα intra-varietal VNI/VNII hybrids and aADα inter-varietal VNII/VNIV hybrids.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>27764108</pmid><doi>10.1371/journal.pone.0163955</doi><tpages>e0163955</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2016-10, Vol.11 (10), p.e0163955-e0163955 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1830671964 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Analysis Antifungal agents Antifungal Agents - pharmacology Biology and Life Sciences Cell size Central nervous system Cryptococcus Cryptococcus gattii Cryptococcus gattii - genetics Cryptococcus gattii - physiology Cryptococcus neoformans Cryptococcus neoformans - genetics Cryptococcus neoformans - physiology Dilution Fungal infections Fungal Proteins - genetics Fungal Proteins - metabolism Fungicides Galleria mellonella Genes, Mating Type, Fungal - genetics Giant cells Haploidy Health aspects Heterozygosity HIV Human immunodeficiency virus Hybridization Hybridization, Genetic - genetics Hybrids Infections Infectious diseases Laboratories Larvae Lysophospholipase - metabolism Mating Mating types Medical research Medical schools Medicine and Health Sciences Melanin Melanins - metabolism Microbial Sensitivity Tests Morphology Moths Multiplex Polymerase Chain Reaction Mycological Typing Techniques Mycoses Pathogens Phospholipase Physical Sciences Reproduction (biology) Research and Analysis Methods Serogroup Sexual reproduction Species Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Virulence Virulence (Microbiology) Virulence - drug effects Virulence - genetics Virulence factors Virulence Factors - analysis Virulence Factors - genetics Virulence Factors - metabolism Yeasts |
| title | Identification and Characterization of VNI/VNII and Novel VNII/VNIV Hybrids and Impact of Hybridization on Virulence and Antifungal Susceptibility Within the C. neoformans/C. gattii Species Complex |
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