Molecular identification, genotypic heterogeneity and comparative pathogenicity of environmental isolates of Free

Abstract

, formerly , is typically isolated from environmental sources, but also occasionally from clinical specimens. Other close relatives may be misidentified as by phenotypic methods. usually lacks melanin; however, melanin-forming strains have also been isolated.

Although melanin production by encapsulated budding yeasts is considered a major virulence factor, the comparative pathogenicity of melanin-forming and non-melanized environmental strains of has rarely been studied.

We performed phenotypic and molecular identification and determined the genotypic heterogeneity among isolates. We also studied the pathogenicity of melanin-forming and non-melanized strains in normal and immunosuppressed mice.

Eleven environmental isolates were tested for their identity by Vitek2 and/or ID32C systems, and by PCR-sequencing of the internal transcribed spacer (ITS) region and D1/D2 domains of ribosomal DNA (rDNA). Genotypic heterogeneity was studied by sequence comparisons. The pathogenicity of melanized and non-melanized strains was studied in intravenously infected normal and immunosuppressed BALB/c mice.

Phenotypic methods identified seven of the environmental isolates, while PCR-sequencing of the ITS region and D1/D2 domains of rDNA detected two and five isolates, respectively, as . Sequence comparisons demonstrated genotypic heterogeneity among . The remaining four environmental isolates yielded expected results. None of the normal mice infected with 10 cells of melanized/non-melanized strains died. Infection of immunosuppressed mice with 10 cells caused higher mortality with non-melanized , while viable counts in brain/lung tissue were higher in mice infected with a melanized strain and were detectable for up to 14 days.

Phenotypic methods lacked specificity, but PCR-sequencing of D1/D2 domains correctly identified and sequence comparisons demonstrated the genotypic heterogeneity of the isolates. Both melanized and non-melanized strains at a higher dose caused mortality in immunosuppressed mice and persisted in brain/lung tissue up to 14 days post-infection.

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2020-09-18
2024-03-29
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