High-Throughput Microsatellite Markers Development for Genetic Characterization of Emerging Sporothrix Species

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Losada, Luiza Chaves de Miranda Leonhardt ; Monteiro, Ruan Campos ; de Carvalho, Jamile Ambrosio ; Hagen, Ferry ; Fisher, Matthew C. ; Spruijtenburg, Bram ; Meis, Jacques F. ; de Groot, Theun ; Goncalves, Sarah Santos ; Negroni, Ricardo ; Kano, Rui ; Bonifaz, Alexandro ; de Camargo, Zoilo Pires ; Rodrigues, Anderson Messias

• Document type: Journal article
• Source: JOURNAL OF FUNGI; v. 9, n. 3, p. 34-pg., 2023-03-01.

Abstract

Sporotrichosis is the main subcutaneous mycosis worldwide transmitted by animal or plant vectors and often escalates to outbreaks or epidemics. The current cat-transmitted sporotrichosis driven by Sporothrix brasiliensis has become a significant public health issue in South America. Transmission dynamics remain enigmatic due to the lack of development of polymorphic markers for molecular epidemiological analysis. This study used a high-throughput mining strategy to characterize simple sequence repeat (SSR) markers from Sporothrix genomes. A total of 118,140-143,912 SSR loci were identified (82,841-98,369 unique markers), with a 3651.55-3804.65 SSR/Mb density and a majority of dinucleotides motifs (GC/CG). We developed a panel of 15 highly polymorphic SSR markers suitable for genotyping S. brasiliensis, S. schenckii, and S. globosa. PCR amplification revealed 240 alleles in 180 Sporothrix isolates with excellent polymorphic information content (PIC = 0.9101), expected heterozygosity (H = 0.9159), and discriminating power (D = 0.7127), supporting the effectiveness of SSR markers in uncovering cryptic genetic diversity. A systematic population genetic study estimated three clusters, corresponding to S. brasiliensis (population 1, n = 97), S. schenckii (population 2, n = 49), and S. globosa (population 3, n = 34), with a weak signature of mixed ancestry between populations 1 and 2 or 3 and 2. Partitioning of genetic variation via AMOVA revealed highly structured populations (phi PT = 0.539; Nm = 0.213; p < 0.0001), with approximately equivalent genetic variability within (46%) and between (54%) populations. Analysis of SSR diversity supports Rio de Janeiro (RJ) as the center of origin for contemporary S. brasiliensis infections. The recent emergence of cat-transmitted sporotrichosis in northeastern Brazil indicates an RJ-Northeast migration resulting in founder effects during the introduction of diseased animals into sporotrichosis-free areas. Our results demonstrated high cross-species transferability, reproducibility, and informativeness of SSR genetic markers, helping dissect deep and fine-scale genetic structures and guiding decision making to mitigate the harmful effects of the expansion of cat-transmitted sporotrichosis. (AU)

• FAPESP's process: 18/21460-3 - Study of different antigenic preparations of Paracoccidioides lutzii for the standardization of the ELISA test as an aid in the diagnosis of paracoccidioidomycosis due to P. lutzii
• Grantee: Zoilo Pires de Camargo
• Support Opportunities: Regular Research Grants
• FAPESP's process: 17/27265-5 - Molecular epidemiology and genomic perspectives on the evolution and spread of emerging fungal pathogens
• Grantee: Anderson Messias Rodrigues
• Support Opportunities: Research Grants - Young Investigators Grants