Easy Storage Strategies for<i>Sporothrix</i>spp. Strains

Sporothrix ( Ophiostomatales ) comprises species that are pathogenic to humans and other mammals as well as environmental fungi. Developments in molecular phylogeny have changed our perceptions about the epidemiology, host-association, and virulence of Sporothrix . The classical agent of sporotrichosis, Sporothrix schenckii , now comprises several species nested in a clinical clade with S. brasiliensis , S. globosa , and S. luriei . To gain a more precise view of outbreaks dynamics, structure, and origin of genetic variation within and among populations of Sporothrix , we applied three sets of discriminatory AFLP markers (#3 EcoRI-GA/MseI-TT, #5 EcoRI-GA/MseI-AG, and #6 EcoRI-TA/MseI-AA) and mating-type analysis to a large collection of human, animal and environmental isolates spanning the major endemic areas. A total of 451 polymorphic loci were amplified in vitro from 188 samples, and revealed high polymorphism information content ( PIC = 0.1765–0.2253), marker index ( MI = 0.0001–0.0002), effective multiplex ratio ( E = 15.1720–23.5591), resolving power ( Rp = 26.1075–40.2795), discriminating power ( D = 0.9766–0.9879), expected heterozygosity ( H = 0.1957–0.2588), and mean heterozygosity ( H avp = 0.000007–0.000009), demonstrating the effectiveness of AFLP markers to speciate Sporothrix . Analysis using the program structure indicated three genetic clusters matching S. brasiliensis (population 1), S. schenckii (population 2), and S. globosa (population 3), with the presence of patterns of admixture amongst all populations. AMOVA revealed highly structured clusters (PhiPT = 0.458–0.484, P < 0.0001), with roughly equivalent genetic variability within (46–48 %) and between (52–54 %) populations. Heterothallism was the exclusive mating strategy, and the distributions of MAT1-1 or MAT1-2 idiomorphs were not significantly skewed (1:1 ratio) for S. schenckii (χ 2 = 2.522; P = 0.1122), supporting random mating. In contrast, skewed distributions were found for S. globosa (χ 2 = 9.529; P = 0.0020) with a predominance of MAT1-1 isolates, and regional differences were highlighted for S. brasiliensis with the overwhelming occurrence of MAT1-2 in Rio de Janeiro (χ 2 = 14.222; P = 0.0002) and Pernambuco (χ 2 ...

show abstract

“…Filamentous colonies were grown on Sabouraud Dextrose Agar (SDA) at 25 °C and co-cultured every fourteen days ( Brilhante et al. 2015 ).…”

Section: Methodsmentioning

confidence: 99%

Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers

Carvalho¹,

Beale²,

Hagen³

et al. 2021

Studies in Mycology

View full text Add to dashboard Cite

show abstract

“…The isolates were identified down to species level by comparison with the descriptions presented by Rodrigues et al . [ 41 – 44 ], and were stored at room temperature in slant cultures on Sabouraud dextrose agar (Difco Laboratories, Detroit, MI, USA) [ 45 ].…”

Section: Methodsmentioning

confidence: 99%

“…This study included 27 Sporothrix isolates (S. brasiliensis, n = 9; S. schenckii, n = 8; S. globosa, n = 6; S. mexicana, n = 2; S. chilensis, n = 1, and S. pallida n = 1), obtained from clinical lesions of patients with varying degrees of disease severity (n = 20), from animals (n = 3) or from environmental sources (n = 4). The isolates were identified down to species level by comparison with the descriptions presented by Rodrigues et al [41][42][43][44], and were stored at room temperature in slant cultures on Sabouraud dextrose agar (Difco Laboratories, Detroit, MI, USA) [45].…”

Section: Fungal Strainsmentioning

confidence: 99%

Genome-wide mapping using new AFLP markers to explore intraspecific variation among pathogenic Sporothrix species

et al. 2020

Self Cite

View full text Add to dashboard Cite

Sporotrichosis is a chronic subcutaneous mycosis caused by Sporothrix species, of which the main aetiological agents are S. brasiliensis, S. schenckii, and S. globosa. Infection occurs after a traumatic inoculation of Sporothrix propagules in mammals' skin and can follow either a classic route through traumatic inoculation by plant debris (e.g., S. schenckii and S. globosa) or an alternative route through zoonotic transmission from animals (e.g., S. brasiliensis). Epizootics followed by a zoonotic route occur in Brazil, with Rio de Janeiro as the epicenter of a recent cat-transmitted epidemic. DNA-based markers are needed to explore the epidemiology of these Sporothrix expansions using molecular methods. This paper reports the use of amplified-fragment-length polymorphisms (AFLP) to assess the degree of intraspecific variability among Sporothrix species. We used whole-genome sequences from Sporothrix species to generate 2,304 virtual AFLP fingerprints. In silico screening highlighted 6 primer pair combinations to be tested in vitro. The protocol was used to genotype 27 medically relevant Sporothrix. Based on the overall scored AFLP markers (97-137 fragments), the values of polymorphism information content (PIC = 0.2552-0.3113), marker index (MI = 0.002-0.0039), effective multiplex ratio (E = 17.8519-35.2222), resolving power (Rp = 33.6296-63.1852), discriminating power (D = 0.9291-0.9662), expected heterozygosity (H = 0.3003-0.3857), and mean heterozygosity (H avp = 0.0001) demonstrated the utility of these primer combinations for discriminating Sporothrix. AFLP markers revealed cryptic diversity in species previously thought to be the most prevalent clonal type, such as S. brasiliensis, responsible for cat-transmitted sporotrichosis, and S. globosa responsible for large sapronosis outbreaks in Asia. Three combinations (#3 EcoRI-FAM-GA/MseI-TT, #5 EcoRI-FAM-GA/MseI-AG, and #6 EcoRI-FAM-TA/MseI-AA) provide the best diversity indices and lowest error rates. These methods make it easier to track routes of disease transmission during epizooties and zoonosis, and our DNA fingerprint assay can be further transferred between laboratories to give insights into the ecology and PLOS NEGLECTED TROPICAL DISEASES

show abstract

“…Colonies that exhibited morphological aspects similar to Sporothrix spp. were subcultured and stocked in glass tubes with water supplemented with chloramphenicol at room temperature for further identification 19 . Eight Sporothrix spp.…”

Section: Methodsmentioning

confidence: 99%

Molecular‐based assessment of diversity and population structure of Sporothrix spp. clinical isolates from Espírito Santo‐Brazil

Rocha

Terra

Oliveira

et al. 2020

Mycoses

Self Cite

View full text Add to dashboard Cite

Background Sporotrichosis is a subcutaneous mycosis caused by Sporothrix species that affects humans and animals. Little information on the genetic diversity and population structure of the pathogen is available for Brazil, which is needed to design effective strategies to tackle the advance of sporotrichosis in endemic areas. Objectives We assessed the genetic diversity and mating‐type distribution of Sporothrix isolates recovered from human and feline cases of sporotrichosis in Espírito Santo‐Brazil to better understand the population structure, epidemiology and diversification of this pathogen, as well as to explore the possible routes of transmission involved in the ongoing outbreaks. Methods In all, 75 Sporothrix isolates were identified with phenotypic characteristics. Then, fungal DNA extraction was performed, and the species‐specific PCR technique was applied, using markers directed to the calmodulin gene. The mating‐type idiomorph of species was identified by PCR using primers targeting the MAT1‐1 and MAT1‐2 loci. Results Among the 75 Sporothrix isolates, 76% were confirmed as S brasiliensis and 24% as S schenckii sensu stricto. S brasiliensis was more prevalent in the metropolitan area and S schenckii s. str. in the mountainous region of the state. In both species, the presence of the two sexual idiomorphs was detected, suggesting that they are heterothallic species. Conclusions Our data suggest that sporotrichosis takes on an epidemic‐urban character involving S brasiliensis. This species in Espírito Santo is likely to originate from Rio de Janeiro, as most isolates harbour the same MAT 1‐2 locus. We confirm that S brasiliensis has significantly broadened its area of occurrence, an essential feature of emerging pathogens.

show abstract

Easy Storage Strategies forSporothrixspp. Strains

Cited by 8 publications

References 16 publications

Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers

Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers

Genome-wide mapping using new AFLP markers to explore intraspecific variation among pathogenic Sporothrix species

Molecular‐based assessment of diversity and population structure of Sporothrix spp. clinical isolates from Espírito Santo‐Brazil

Contact Info

Product

Resources

About