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

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 dyn...

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Autores principales: Losada, Luiza Chaves de Miranda Leonhardt, Monteiro, Ruan Campos, de Carvalho, Jamile Ambrósio, Hagen, Ferry, Fisher, Matthew C., Spruijtenburg, Bram, Meis, Jacques F., de Groot, Theun, Gonçalves, Sarah Santos, Negroni, Ricardo, Kano, Rui, Bonifaz, Alexandro, de Camargo, Zoilo Pires, Rodrigues, Anderson Messias
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10054832/
https://www.ncbi.nlm.nih.gov/pubmed/36983522
http://dx.doi.org/10.3390/jof9030354
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author Losada, Luiza Chaves de Miranda Leonhardt
Monteiro, Ruan Campos
de Carvalho, Jamile Ambrósio
Hagen, Ferry
Fisher, Matthew C.
Spruijtenburg, Bram
Meis, Jacques F.
de Groot, Theun
Gonçalves, Sarah Santos
Negroni, Ricardo
Kano, Rui
Bonifaz, Alexandro
de Camargo, Zoilo Pires
Rodrigues, Anderson Messias
author_facet Losada, Luiza Chaves de Miranda Leonhardt
Monteiro, Ruan Campos
de Carvalho, Jamile Ambrósio
Hagen, Ferry
Fisher, Matthew C.
Spruijtenburg, Bram
Meis, Jacques F.
de Groot, Theun
Gonçalves, Sarah Santos
Negroni, Ricardo
Kano, Rui
Bonifaz, Alexandro
de Camargo, Zoilo Pires
Rodrigues, Anderson Messias
author_sort Losada, Luiza Chaves de Miranda Leonhardt
collection PubMed
description 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 (ΦPT = 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.
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spelling pubmed-100548322023-03-30 High-Throughput Microsatellite Markers Development for Genetic Characterization of Emerging Sporothrix Species Losada, Luiza Chaves de Miranda Leonhardt Monteiro, Ruan Campos de Carvalho, Jamile Ambrósio Hagen, Ferry Fisher, Matthew C. Spruijtenburg, Bram Meis, Jacques F. de Groot, Theun Gonçalves, Sarah Santos Negroni, Ricardo Kano, Rui Bonifaz, Alexandro de Camargo, Zoilo Pires Rodrigues, Anderson Messias J Fungi (Basel) Article 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 (ΦPT = 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. MDPI 2023-03-14 /pmc/articles/PMC10054832/ /pubmed/36983522 http://dx.doi.org/10.3390/jof9030354 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Losada, Luiza Chaves de Miranda Leonhardt
Monteiro, Ruan Campos
de Carvalho, Jamile Ambrósio
Hagen, Ferry
Fisher, Matthew C.
Spruijtenburg, Bram
Meis, Jacques F.
de Groot, Theun
Gonçalves, Sarah Santos
Negroni, Ricardo
Kano, Rui
Bonifaz, Alexandro
de Camargo, Zoilo Pires
Rodrigues, Anderson Messias
High-Throughput Microsatellite Markers Development for Genetic Characterization of Emerging Sporothrix Species
title High-Throughput Microsatellite Markers Development for Genetic Characterization of Emerging Sporothrix Species
title_full High-Throughput Microsatellite Markers Development for Genetic Characterization of Emerging Sporothrix Species
title_fullStr High-Throughput Microsatellite Markers Development for Genetic Characterization of Emerging Sporothrix Species
title_full_unstemmed High-Throughput Microsatellite Markers Development for Genetic Characterization of Emerging Sporothrix Species
title_short High-Throughput Microsatellite Markers Development for Genetic Characterization of Emerging Sporothrix Species
title_sort high-throughput microsatellite markers development for genetic characterization of emerging sporothrix species
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10054832/
https://www.ncbi.nlm.nih.gov/pubmed/36983522
http://dx.doi.org/10.3390/jof9030354
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