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
Laboratory of Emerging Fungal Pathogens, Department of Microbiology, Immunology, and Parasitology, Discipline of Cellular Biology, Federal University of São Paulo (UNIFESP), São Paulo 04023062, Brazil.
Department of Medicine, Discipline of Infectious Diseases, Federal University of São Paulo (UNIFESP), São Paulo 04023062, Brazil.
J Fungi (Basel). 2023 Mar 14;9(3):354. doi: 10.3390/jof9030354.
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 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 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 , and . PCR amplification revealed 240 alleles in 180 isolates with excellent polymorphic information content ( = 0.9101), expected heterozygosity ( = 0.9159), and discriminating power ( = 0.7127), supporting the effectiveness of SSR markers in uncovering cryptic genetic diversity. A systematic population genetic study estimated three clusters, corresponding to (population 1, n = 97), (population 2, n = 49), and (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; < 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 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.
孢子丝菌病是全球主要的皮下真菌病，通过动物或植物媒介传播，常演变为暴发或流行。目前由猫传播的孢子丝菌病已成为南美洲一个重大的公共卫生问题。由于缺乏用于分子流行病学分析的多态性标记，传播动态仍然不明。本研究采用高通量挖掘策略，从基因组中鉴定简单序列重复（SSR）标记。共鉴定出118,140 - 143,912个SSR位点（82,841 - 98,369个独特标记），SSR密度为3651.55 - 3804.65个/Mb，且大多数为二核苷酸基序（GC/CG）。我们开发了一组15个高度多态的SSR标记，适用于对、和进行基因分型。PCR扩增在180个分离株中揭示了240个等位基因，具有优异的多态信息含量（= 0.9101）、期望杂合度（= 0.9159）和鉴别力（= 0.7127），支持SSR标记在揭示隐秘遗传多样性方面的有效性。一项系统的群体遗传学研究估计出三个聚类，分别对应（群体1，n = 97）、（群体2，n = 49）和（群体3，n = 34），群体1与2或3与2之间有微弱的混合血统特征。通过分子方差分析（AMOVA）对遗传变异进行划分，结果显示群体结构高度分化（ΦPT = 0.539；Nm = 0.213；< 0.0001），群体内部（46%）和群体之间（54%）的遗传变异大致相当。对SSR多样性的分析支持里约热内卢（RJ）作为当代感染的起源中心。巴西东北部近期出现的由猫传播的孢子丝菌病表明存在从RJ向东北部的迁移，导致在将患病动物引入无孢子丝菌病地区时产生奠基者效应。我们的结果证明了SSR遗传标记具有高跨物种转移性、可重复性和信息性，有助于剖析深层次和精细尺度的遗传结构，并指导决策以减轻由猫传播的孢子丝菌病扩散带来的有害影响。
