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This study aims to examine the range of genetic variations, assess molecular epidemiology significance, and explore potential modes of transmission of the Sporothrix species associated with the current sporotrichosis outbreaks in Esp\u00edrito Santo, Brazil. In this investigation, 262 samples were evaluated, including 142 from humans and 120 from felines, collected between 2016 and 2021. The isolates were identified based on morphological and molecular characteristics. Sexual idiomorphs were determined by mating-type PCR using primers specific to the MAT1-1 and MAT1-2 loci. Amplified fragment length polymorphism (AFLP) was employed to assess the genetic variability of Sporothrix spp. Finally, antifungal susceptibility testing was performed following the CLSI M38-A2 protocol. Of the 142 human samples, 125 were identified as S. brasiliensis and 17 as S. schenckii s. str. The presence of S. brasiliensis was overwhelming (100%) during outbreaks, highlighting the significant role of domestic cats in the emergence of this species. Heterothallism was the only observed mating strategy. However, the MAT1-2 idiomorph was predominant in cases of cat-transmitted sporotrichosis (\u03c72 = 202.976; p &lt; 0.0001). Our AFLP results show significant intraspecific variability observed among S. brasiliensis isolates in Esp\u00edrito Santo. Different genotypes forming subgroups within the same population suggest that these isolates do not originate from a single ancestor, indicating multiple emergences. Furthermore, terbinafine was the antifungal with the best results in vitro. However, in clinical practice, itraconazole remains the primary treatment choice. Sporotrichosis continues to advance in the state; therefore, the health system must outline one-health strategies to contain the disease to prevent future epidemics.<\/jats:p>","DOI":"10.3390\/jof9080831","type":"journal-article","created":{"date-parts":[[2023,8,7]],"date-time":"2023-08-07T06:38:48Z","timestamp":1691390328000},"page":"831","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Human and Feline Sporotrichosis in a Reference Center of Southeastern Brazil: Genetic Differentiation, Diversity, and Antifungal Susceptibility of Sporothrix Species"],"prefix":"10.3390","volume":"9","author":[{"given":"Sarah Santos","family":"Gon\u00e7alves","sequence":"first","affiliation":[{"name":"Infectious Diseases Postgraduate Program, Center for Research in Medical Mycology, Department of Pathology, Federal University of Esp\u00edrito Santo (UFES), Vitoria 29043900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabela","family":"da Cruz Bahiense Rocha","sequence":"additional","affiliation":[{"name":"Infectious Diseases Postgraduate Program, Center for Research in Medical Mycology, Department of Pathology, Federal University of Esp\u00edrito Santo (UFES), Vitoria 29043900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4023-1415","authenticated-orcid":false,"given":"Bruno Carneiro","family":"Rediguieri","sequence":"additional","affiliation":[{"name":"Infectious Diseases Postgraduate Program, Center for Research in Medical Mycology, Department of Pathology, Federal University of Esp\u00edrito Santo (UFES), Vitoria 29043900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jamile Ambr\u00f3sio","family":"de Carvalho","sequence":"additional","affiliation":[{"name":"Laboratory of Emerging Fungal Pathogens, Department of Microbiology, Immunology and Parasitology, Discipline of Cellular Biology, Federal University of S\u00e3o Paulo (UNIFESP), S\u00e3o Paulo 04023062, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simone Bravim","family":"Maifrede","sequence":"additional","affiliation":[{"name":"Center for Research in Medical Mycology, Department of Pathology, Federal University of Esp\u00edrito Santo (UFES), Vitoria 29043900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wdson Luis Lima","family":"Kruschewsky","sequence":"additional","affiliation":[{"name":"Department of Infectious and Parasitic Diseases, The Clinical Hospital of the Faculty of Medicine of the University of S\u00e3o Paulo, S\u00e3o Paulo 05403010, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alo\u00edsio","family":"Falqueto","sequence":"additional","affiliation":[{"name":"Department of Medical Clinic, Cassiano Ant\u00f4nio Moraes University Hospital (HUCAM), Federal University of Esp\u00edrito Santo (UFES), Vitoria 29043900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0891-4563","authenticated-orcid":false,"given":"Anderson Messias","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Laboratory of Emerging Fungal Pathogens, Department of Microbiology, Immunology and Parasitology, Discipline of Cellular Biology, Federal University of S\u00e3o Paulo (UNIFESP), S\u00e3o Paulo 04023062, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,8,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"De Carolis, E., Posteraro, B., and Sanguinetti, M. 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