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Sporotrichosis is the most important subcutaneous mycosis that affects humans and animals worldwide. The mycosis is caused after a traumatic inoculation of fungal propagules into the host and may follow an animal or environmental transmission route. The main culprits of sporotrichosis are thermodimorphic Sporothrix species embedded in a clinical clade, including S. brasiliensis, S. schenckii, S. globosa, and S. luriei. Although sporotrichosis occurs worldwide, the etiological agents are not evenly distributed, as exemplified by ongoing outbreaks in Brazil and China, caused by S. brasiliensis and S. globosa, respectively. The gold standard for diagnosing sporotrichosis has been the isolation of the fungus in vitro. However, with the advance in molecular techniques, molecular assays have complemented and gradually replaced the classical mycological tests to quickly and accurately detect and/or differentiate molecular siblings in Sporothrix. Nearly all techniques available for molecular diagnosis of sporotrichosis involve PCR amplification, which is currently moving towards detecting Sporothrix DNA directly from clinical samples in multiplex qPCR assays. From an epidemiological perspective, genotyping is key to tracing back sources of Sporothrix infections, detecting diversity in outbreak areas, and thus uncovering finer-scale epidemiological patterns. Over the past decades, molecular epidemiological studies have provided essential information to policymakers regarding outbreak management. From high-to-low throughput genotyping methods, MLSA, AFLP, SSR, RAPD, PCR-RFLP, and WGS are available to assess the transmission dynamics and sporotrichosis expansion. This review discusses the trends in the molecular diagnosis of sporotrichosis, genotyping techniques applied in molecular epidemiological studies, and perspectives for the near future.

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.

This study evaluated in vitro activity of ethanol extract, fractions, and isolated substance from Amazon species against promastigotes of L. amazonensis. The ethanol extracts were concentrated and fractionation. The anti-promastigote activity was evaluated through the cell viability assessment method (MTT). The ethanol extract, fractions, and isolated substance from Himatanthus articulatus and Parahancornia fasciculata were inactive in promastigote of L. amazonensis, as the ethanol extract of Physalis angulata. The hexane fractions from different parts of Montrichardia linifera showed anti-promastigote activity probably due to the presence of steroids and terpenes. All species in studies were inactive, except of M. linifera. The few polar constituents can be responsible for the activity. Therefore, the isolation and purification of the active on L. amazonensis promastigotes are urgently required.

Purpose The purpose of this study was to investigate the diversity of filamentous fungi and the hydrolytic potential of their enzymes for a future understanding of the influence of these factors on the sensory characteristics of the cocoa beans used to obtain chocolate. Methods Filamentous fungi were isolated from the natural cocoa fermentation boxes in the municipality of Tucuman, Pará, Brazil, and evaluated for the potential production of amylases, cellulases, pectinases, and xylanases. The fermentation was monitored by analyzing the pH and temperature. The strains were identified by sequencing the ITS1/ITS4 section of the 5.8S rDNA and partially sequencing the 18S and 28S regions, and the molecular identification was confirmed by phylogenetic reconstruction. Result The fungi isolated were comprised of three classes from the Ascomycota phylum and one class from the Basidiomycota phylum. There were found 19 different species, of this amount 16 had never been previously reported in cocoa fermentation. This fact characterizes the fermentation occurring in this municipality as having wide fungal diversity. Most of the strains isolated had the ability to secrete enzymes of interest. Cladosporium cladosporioides, Fomitopsis subtropical, Aspergillus versicolor, Penicillium pimiteouiense, Phanerochaete australis, Neonothopanus nambi, and Aspergillus parasiticus were the strains that excelled in the secretion of the following enzymes: amylase, pectinase, cellulase, and xylanase. Conclusion The presence of 16 species not yet reported in cocoa seed fermentations and their potential hydrolytic activities show a diversity of filamentous fungi in this microbial biome that needs to be better understood.

The most prominent historical buildings in Belém do Pará (Northern Brazil) have modernist stained-glass windows, which were commissioned from Europe since the end of the 19 th century. Some of them present biodegradation; however, there is no information about the microbial activity on them. The present work is focused on the biodeterioration by fungi on some of these Modern stained-glass windows. The fungal communities were collected, isolated and then identified by means of molecular methods. Additionally, a laboratory-based biodeterioration experiment was carried out to assess the fungal activity on replica glass samples with three different chemical compositions. The replica samples were inoculated with a four-fungal species mixture and incubated under optimal growth conditions for 5 months. Optical microscopy, µ-PIXE, SEM-EDS and FTIR-ATR were performed to evaluate the biodeterioration of the soda-lime silicate glasses. This multidisciplinary approach showed that the inoculated spores (Aspergillus arenarioides, Fusarium oxysporum, Hortaea werneckii, and Trichoderma longibrachiatum) were able to form substantial mycelia in all replica glass samples. The main alterations observed were small crystals, hyphae fingerprints and a slight decrease on the glass surface smoothness. Despite the aforementioned damages, the soda-lime silicate glass compositions showed high resistance against the inoculated fungal species.

Sporotrichosis is a subcutaneous mycosis that affects animals and humans. Varying in severity, occurrences range from local lesions to systemic involvement. It is caused by thermodimorphic and saprobic fungi from the Sporothrix pathogenic clade. This study aimed to identify the species and the sexual idiomorph distribution patterns responsible for diagnosed cases of sporotrichosis in São José do Rio Preto, Brazil. We included 188 isolates of Sporothrix sp. from feline lesions and 27 of human origin, which underwent molecular identification and genotyping for mating-type MAT1-1 and MAT1-2. The results showed that Sporothrix brasiliensis is the prevalent species in feline sporotrichosis outbreaks with the overwhelming presence of a single mating-type, MAT1-2 (P &lt; 0.0001), suggesting a prevalently clonal form of spread. Morphological analyses did not discriminate among cryptic species in the genus Sporothrix, and molecular identification was essential for the correct identification of the species responsible for the observed cases of sporotrichosis. Distribution analyses of MAT1-2 isolates support the hypothesis of unidirectional migration from the current epidemics in São Paulo and Rio de Janeiro to the municipality of São José do Rio Preto.