MEM INST OSWALDO CRUZ, RIO DE JANEIRO, 108(7) November 2013
PAGES: 936-938 DOI: 10.1590/0074-0276130239 Short communication
Molecular identification of Sporothrix species involved in the first familial outbreak of sporotrichosis in the state of Espírito Santo, southeastern Brazil

Manoel Marques Evangelista Oliveira1,+, Simone Bravim Maifrede2, Mariceli Araújo Ribeiro2, Rosely Maria Zancope-Oliveira1

1Laboratório de Micologia, Instituto de Pesquisa Clínica Evandro Chagas-Fiocruz, Rio de Janeiro, RJ, Brasil
2Núcleo de Doenças Infecciosas, Universidade Federal do Espírito Santo, Vitória, ES, Brasil

Abstract

Sporotrichosis is a widespread subcutaneous mycosis caused by the dimorphic fungi now known as the Sporothrix schenckii complex. This complex is comprised of at least six species, including Sporothrix albicans, Sporothrix brasiliensis, Sporothrix globosa, Sporothrix luriei, Sporothrix mexicana and S. schenckii. Cases of sporotrichosis have significantly increased in Brazil over the past decade, especially in the state of Rio de Janeiro (RJ), where an epidemic among cat owners has been observed. The zoonotic transmission from cats to humans suggests a common source of infection and indicates that animals can act as vectors. We performed a molecular characterisation of samples collected during the first outbreak of familial sporotrichosis caused by S. brasiliensis in the state of Espírito Santo, Brazil. These results represent the first description of such an outbreak outside the endemic area of zoonotic sporotrichosis in RJ.

Sporotrichosis is a widespread subcutaneous mycosis with high endemicity in Latin America, South Africa, India and Japan (Lopez-Romero et al. 2011). The infection is caused by dimorphic fungi of the Sporothrix schenckii species complex (Marimon et al. 2007), which is comprised of at least six species, including Sporothrix albicans, Sporothrix brasiliensis, Sporothrix globosa, Sporothrix luriei, Sporothrix mexicana and Sporothrix schenckii (Marimon et al. 2007, 2008). In humans, sporotrichosis is classically known as gardener's disease and is generally associated with soil transmission. In the sporotrichosis epidemic that occurred in the state of Rio de Janeiro (RJ), Brazil, transmission of the disease was associated with scratches or bites from cats infected with S. schenckii (Schubach et al. 2008).

A species-level classification of the Sporothrix complex has now been proposed (Marimon et al. 2007). In a previous study, Oliveira et al. (2011) disagreed with earlier data published by Marimon et al. (2007) that assumed that the differentiation of species within the Sporothrix complex could be easily accomplished without molecular methods. Our group has reported that the correlation between molecular data and phenotypic characteristics is crucial for the identification of species in the Sporothrix complex. In the current study, we characterised four Sporothrix strains at the species level using a polyphasic analysis (Oliveira et al. 2011). The strains were obtained from sporotrichosis cases that occurred in one cat (ES213) and three family members (ES210, ES211 and ES212; ages 30, 14 and 10 years, respectively) from a rural area in the state of Espírito Santo (ES). The patients developed skin ulcers with irregular borders and satellite microabscesses located in the lower buttocks, thighs and neck as clinically and epidemiologically described by Falqueto et al. (2012). The isolated strains were compared with the following three control strains: S. brasi-liensis (CBS 120339; formerly IPEC 16490) (Marimon et al. 2007), S. schenckii (IPEC 27722) (Oliveira et al. 2011) and S. globosa (IPEC 27135) (Oliveira et al. 2011).

The phenotypic characteristics of the isolates are shown in the Table. Overall, the sympodial conidia of the isolates were hyaline or slightly pigmented. The greatest fungal growth was observed at 30ºC with a minimum colony diameter of 37 mm and a maximum colony diameter of 45 mm. At 37ºC, a colony diameter minimum of 9 mm and a colony diameter maximum of 13 mm were observed.

Assimilation tests using dextrose, sucrose and raffinose were performed in triplicate in yeast nitrogen base medium. After 10 days, the four isolates assimilated only sucrose and glucose (Table). All isolates showed a biochemical pattern of carbohydrate assimilation typical of S. globosa or S. albicans according to Marimon's key. However, the average colony diameter at 30ºC (diameter of the colonies not exceeding 50 mm) excluded S. albicans in this cases; in addition the strains produce dematiaceous conidia. The isolates also could not be classified as S. globosa because they were thermotolerant. The isolates were therefore presumptively identified as Sporothrix spp based on phenotypic characteristics.

Genomic DNA was obtained from the isolates in mould phase and sequencing of the partial calmodulin-encoding gene (CAL) was performed as previously described (Oliveira et al. 2010, 2011) using the Genomic Platform-DNA Sequencing at Oswaldo Cruz Foundation (RPT01A) in Brazil. The similarity of the partial CAL gene sequences to those obtained from the National Center for Biotechnology Information GenBank database confirmed with high bootstrap support that all isolates were S. brasiliensis (Figure). All sequences were deposited in the GenBank database under accessions JQ915210 through JQ915213.

 

 

During the past decade, cases of sporotrichosis have significantly increased in Brazil. The occurrence of these cases has been particularly evident in RJ; here, an epidemic resulting from zoonotic transmission from cats to humans has been observed, leading to small outbreaks among cats owners and veterinary professionals. Such an outbreak suggests a common source of infection between humans and cats and indicates that animals can act as vectors for the transmission of this fungal disease (Schubach et al. 2008, Reis et al. 2009).

All isolates were obtained from individuals living in a rural area in ES. ES borders RJ, which is endemic to zoonotic sporotrichosis (Falqueto et al. 2012). The outbreak in ES showed similarities to cases of epidemic sporotrichosis reported in RJ, including the fact that most individuals affected were women that were engaged in domestic duties, in contact with sick cats and were from low-income areas (Freitas et al. 2010). These similarities suggest that this emerging disease could be spreading in Brazil.

To our knowledge, this is the first molecular characterisation of a familial outbreak of zoonotic sporotrichosis caused by S. brasiliensis outside of the endemic area in RJ.

 

REFERENCES

Falqueto A, Maifrede SB, Ribeiro MA 2012. Unusual clinical presentation of sporotrichosis in three members of one family. Int J Dermatol 51: 434-438.

Freitas DF, do Valle AC, Almeida-Paes R, Bastos FI, Galhardo MC 2010. Zoonotic sporotrichosis in Rio de Janeiro, Brazil: a protracted epidemic yet to be curbed. Clin Infect Dis 50: 453.

Lopez-Romero E, Reyes-Montes MR, Perez-Torres A, Ruiz-Baca E, Villagomez-Castro JC, Mora-Montes HM, Flores-Carreón A, Toriello C 2011. Sporothrix schenckii complex and sporotrichosis, an emerging health problem. Future Microbial 6: 85-102.

Marimon R, Cano J, Gene J, Sutton DA, Kawasaki M, Guarro J 2007. Sporothrix brasiliensis, S. globosa and S. mexicana, three new Sporothrix species of clinical interest. J Cllin Microbiol 45: 3198-3206.

Marimon R, Gene J, Cano J, Guarro J 2008. Sporothrix luriei: a rare fungus from clinical origin. Med Mycol 46: 621-625.

Oliveira MME, Almeida-Paes R, Muniz MM, Barros MBL, Ga-lhardo MC, Zancope-Oliveira RM 2010. Sporotrichosis caused by Sporothrix globosa in Rio de Janeiro, Brazil: case report. Mycopathologia 169: 359-363.

Oliveira MME, Almeida-Paes R, Muniz MM, Gutierrez-Galhardo MC, Zancope-Oliveira RM 2011. Phenotypic and molecular identification of Sporothrix isolates from an epidemic area of sporotrichosis in Brazil. Mycopathologia 172: 257-267.

Reis RS, Almeida-Paes R, Muniz MM, Tavares PMS, Monteiro PCF, Schubach TMP, Gutierrez-Galhardo MC, Zancopé-Oliveira RM 2009. Molecular characterisation of Sporothrix schenckii isolates from humans and cats involved in the sporotrichosis epidemic in Rio de Janeiro, Brazil. Mem Inst Oswaldo Cruz 104: 769-774.

Schubach A, Barros MB, Wanke B 2008. Epidemic sporotrichosis. Curr Opin Infect Dis 21: 129-133.

 

Received 19 February 2013
Accepted 13 June 2013
MMEO was supported by CAPES (2445/11-5), RMZO was supported
in part by CNPq (350338/2000-0).
+ Corresponding author: This e-mail address is being protected from spambots. You need JavaScript enabled to view it.

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