Mycopathologia DOI 10.1007/s11046-014-9734-8

Sporothrix schenckii sensu stricto Isolated from Soil in an Armadillo’s Burrow Anderson Messias Rodrigues • Eduardo Bagagli • Zoilo Pires de Camargo • Sandra de Moraes Gimenes Bosco

Received: 15 January 2014 / Accepted: 17 February 2014 Ó Springer Science+Business Media Dordrecht 2014

Abstract Sporotrichosis is a polymorphic disease of man and animals caused by traumatic implantation of propagules into the skin and subcutaneous tissue. Pathogenic species includes S. brasiliensis, S. schenckii, S. globosa and S. luriei. The disease is remarkable for its occurrence as sapronoses and/or zoonosis outbreaks in tropical and subtropical areas; although, the ecology of the clinical clade is still puzzling. Here, we describe an anamorphic Sporothrix strain isolated from soil in an armadillo’s burrow, which was located in a hyper endemic area of Paracoccidioidomycosis in Brazil. This isolate was identified as S. schenckii sensu stricto (Clade IIa) based on morphological and physiological characteristics and phylogenetic analyses of calmodulin sequences. We then discuss the role of the nine-banded armadillo Dasypus novemcinctus as a natural carrier of Sporothrix propagules to better understand Sporothrix sources in nature and reveal essential aspects about the pathogen’s eco-epidemiology.

A. M. Rodrigues  Z. P. de Camargo Disciplina de Biologia Celular, Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de Sa˜o Paulo, Sa˜o Paulo, SP 04023-062, Brazil E. Bagagli  S. de Moraes Gimenes Bosco (&) Departamento de Microbiologia e Imunologia, Instituto de Biocieˆncias de Botucatu, Universidade Estadual Paulista (UNESP), Campus Botucatu, Botucatu, SP 18618-970, Brazil e-mail: [email protected]

Keywords Armadillo  Sporothrix schenckii  Soil  Sporotrichosis  Eco-epidemiology  Ecology

Introduction The species in the genus Sporothrix have a wide range of ecological niches and multiple reservoirs in the environment. This ecological diversity directly reflects the eco-epidemiology, distribution, and incidence of the Sporothrix species. The infectious disease resulting from interactions between microorganisms in the genus Sporothrix and vertebrate hosts is called sporotrichosis and leads to damage and propagation in host tissue. This polymorphic disease is associated with a wide variety of warm-blooded animals such as cats, dogs, armadillos, and humans [1, 2]. For a century, sporotrichosis has been attributed to a single microorganism, S. schenckii sensu lato (s.l.), the most important pathogen in the order Ophiostomatales. Molecular tools reveal that S. schenckii s.l. comprises a complex of cryptic pathogenic species phylogenetically related [3]. New species have been proposed based on the nucleotide sequence analysis of protein-coding loci (chitin synthase, b-tubulin, and calmodulin) and phenotypic features. Currently, medically relevant Sporothrix spp. are S. brasiliensis (Clade I), S. schenckii sensu stricto (s. str.) (Clade II), S. globosa (Clade III), and S. luriei (Clade VI) [4]; while, S. mexicana (Clade IV) and S. pallida (Clade V)

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are remotely related in phylogenetic analysis [1, 3–7] and, therefore, are considered apart from the clinical complex [4]. The main differences among species include virulence [8, 9], geographic distribution [3–5], and sensitivity to antifungals [10], which reinforces the importance of the correct species identification. In nature, at 25 °C, Sporothrix spp. probably grows as mycelia and produce conidia, which are responsible for survival and dispersal in the environment. Propagules present in nature are the major source of contamination of patients, who are traumatically inoculated in tropical and subtropical zones. Soil is considered a great reservoir for major pathogenic and non-pathogenic fungal species. Although sporotrichosis can reach epidemic proportions, the natural ecology of pathogenic species in the clinical clade is poorly understood [1, 4]. Distant relatives in the order Ophiostomatales are mainly associated with bark beetles on woody plants [11]. Sporothrix spp. could leave the saprophytic stage and infect a vertebrate host, causing disease when it undergoes a thermo dimorphic transition at 35–37 °C to a yeast-like phase, which is the parasitic form. Epidemiological studies using skin tests with sporotrichin, an antigenic preparation derived from Sporothrix spp., demonstrated that individuals are exposed naturally to Sporothrix spp. in nature [12, 13]. This sensitization is probably due to Sporothrix spp. propagules present in the soil and might occur during manipulation or contact with contaminated soil. Understanding fungus’ habitats could provide clues to the diversity and biological cycle of the S. schenckii complex found in soil. Here, we describe the isolation of S. schenckii s. str. (Clade II) associated with soil recovered from a nine-banded armadillo’s (Dasypus novemcinctus) burrow in Brazil.

Materials and Methods Environmental Samples Soil samples (a total of 25) were collected from the interior and proximity of six different armadillo’s burrows in Botucatu (22°530 2500 S and 48°270 1900 W) and Manduri (23°00 1000 S and 49°190 2900 W) counties, Sa˜o Paulo, Brazil. These samples were initially collected and analyzed to isolate Paracoccidioides spp., since this area is a hyper endemic area of paracoccidioidomycosis [14];

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however, the occurrence of other thermodimorphic agents were also regarded. The samples were weighed in 250-mL Erlenmeyer sterile flasks and added to a sterile saline solution containing G penicillin (1,000 IU/mL), streptomycin sulfate (1.0 mg/mL), and 0.001 % Tween 80. The soil/saline ratio was 1:25. The solution was strongly shaken for 1 min and then allowed to settle for 20 min. Soil sample suspensions were inoculated into the peritoneum of male Swiss BALB-c mice (Mus musculus) and the testis of male golden Syrian hamsters (Mesocricetus auratus), using 1 mL and 0.2 mL, respectively. Animals were euthanized 6 weeks post-inoculation and small fragments of the liver, spleen, testis, and lymph nodes of hamsters and liver and spleen of mice were cultured onto Mycosel agar (Difco)-containing gentamicin (50 lg/mL). Histopathological analyses were also carried out with periodic acid-Schiff (PAS) and Gomori stains. Suspected Sporothrix cultures were identified down to species level according to Marimon et al. [5, 6]. The phenotypic parameters assayed include vegetative growth on potato dextrose agar (PDA, Oxoid) at different temperatures (30, 35, 37, and 40 °C), the colony colors on corn meal agar (CMA, Oxoid), the assimilation profile of carbon sources (raffinose, ribitol, and sucrose), and morphological microscopic features of in vitro cultivations. The S. schenckii s. str. strain CBS 359.36 was used for phenotypic comparisons. Molecular Characterization For molecular characterization, DNA was extracted and purified directly from fungal mycelial colonies with the Fast DNA kit (MP Biomedicals, USA) according to the manufacturer’s protocol. An amplicon around 800 bp, the calmodulin locus, was amplified from genomic DNA using the primer set CL1 and CL2A, as described by O’Donnell et al. [15]. Sequences were aligned using SATe´-II software [16] and neighbor-joining and maximum likelihood phylogenetic analyses were performed using Mega5 software [17] to determine the clade position. Genetic distances were calculated for all pairwise comparisons of the isolate, according to the Kimura 2-parameters [18]. Susceptibility Test We characterized the response to antifungal agents using an in vitro microdilution susceptibility test [10] with six antifungal drugs: voriconazole, fluconazole,

Mycopathologia

Fig. 1 Hamster 6 weeks after inoculation with a soil sample positive for S. schenckii. a An ulcerated skin lesion in the scrotum; b ulcerated skin lesions and swelling of the joint in the hind limb

itraconazole, amphotericin B, 5-fluorocytosine, and caspofungin. These assays were performed according to Clinical and Laboratory Standards Institute guidelines; document M38-A2.

Results We did not recover any culture morphologically similar to Sporothrix spp. after direct plating of soil samples on Mycosel what suggest that recovery from certain sites can still be challenging. A total of 122 mice and 92 hamsters were evaluated, and Paracoccidioides brasiliensis was not present in any soil sample, based on animal inoculation, even when plating a considerable number of tissue fragments. Despite the absence of Paracoccidioides spp. [14], colonies resembling Sporothrix spp. were present in cultures of the testis, spleen, liver, and mesenteric lymph node of 6 hamsters and the spleen and liver of 2 mice inoculated with a sample collected at the interior (0.6–0.8 m depth) of an armadillo burrow in Manduri County. The inoculated mice did not show any abnormalities during an external examination; while, the hamsters had very swollen testis and ulcerated lesions on the skin of the scrotum. It was also observed that all joints of the former and hind limbs were swollen with small ulcerated lesions of the skin (Fig. 1). During the necropsies, lesions were not present in mice; although, there was discrete hepatosplenomegaly. Hamsters had ulcerated testes with

orchitis containing caseous necrosis as well as hepatosplenomegaly. Histopathological sections showed great numbers of budding yeast cells, a characteristic of pathogenic species in the S. schenckii complex (Fig. 2). The macroscopic morphology characteristics of this environmental isolate (Ss61) were similar to the type strain of S. schenckii (CBS 359.36). After 21 days of incubation at 30 °C, colonies on PDA had a diameter of 49.31 ± 0.17 mm; colonies were moist, with a glabrous surface, and cream to dark brown in color. Microscopically, the isolate had thin, septated hyaline hyphae and formed spores on PDA after 8–12 days at 30 °C. Then, intercalary or terminal conidial clusters were formed by sympodial growth on conidiophores. The thermal dimorphism switching was obtained after several subcultures in BHI (brain heart infusion) agar and incubation at 37 °C. A cream yeast-like colony was observed. After 21 days of incubation at 37 °C, the isolate growth had a colony diameter of 5.16 ± 0.02 mm. Yeast cells were round to ovoid with budding. The microorganism growth was inhibited 88 and 88.9 % in diameter at 35 and 37 °C, respectively, and it did not grow at 40 °C. Carbohydrate assimilation tests were run in triplicate, and the fungus assimilated glucose, raffinose, ribitol, and saccharose as the only source of carbon. Thus, using morphological and physiological techniques, this isolate was classified as S. schenckii (Fig. 3). The phenotypic data were confirmed by molecular biology using the nucleotide sequence of the calmodulin

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Fig. 2 Histopathological section of an articular lesion from a hamster inoculated with a soil sample positive for S. schenckii showing a great number of yeast cells. a Gomori-Grocott, b PAS, X400

amplicon (KF561244). A BLAST search revealed that our isolate showed 99 % similarity with a Peruvian isolate of S. schenckii s. str., FMR 8608 (AM11744). In addition, using a phylogenetic approach to analyze the calmodulin sequence, this isolate clustered together with those belonging to Clade IIa, which included the S. schenckii s. str. isolates from South America. Phylogenetic analyses, using neighbor-joining and maximum likelihood, showed strict concordance (Icong = 3.69; P value = 4.31 9 10-21) [19], and the topology of our isolate was supported by high bootstrap values (98/94). Apart from itraconazole, the antifungal drugs tested did not significantly affect the environmental S. schenckii s. str. strain studied. The minimum inhibitory concentrations (MICs) for the drugs after 72 h were 16 lg/mL for voriconazole; [64 lg/mL for fluconazole; 1 lg/mL for itraconazole; 16 lg/mL for amphotericin B; 64 lg/mL for 5-fluorocytosine; and [16 lg/mL for caspofungin.

Discussion Given the sudden emergence of Sporothrix species as agents of sapronoses and/or zoonosis, studies dealing with source, habitat, and ecological niche are overdue. In Brazil, very little information is available on soil being a source of human fungal infection [20]. The first report of a pathogenic fungus isolated from Brazilian soil was described by Silva [21] in 1956, when Histoplasma capsulatum was isolated in an endemic area of visceral leishmaniasis. To our knowledge, there are few reports of isolations of S. schenckii s. str. from soil in Brazil; yet, the rate of

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infection is increasing in South America [1, 2], and soil and many types of vegetative material represent the major sources of S. schenckii infection [22–24]. Several attempts were made to obtain environmental samples of Sporothrix species in endemic areas [13, 22, 25–28]; although, there was low positivity from soil samples, similar to this study. This may be related to the sampling conditions, seasonality, and methodology. Mackinnon et al. [29] isolated a strain of S. schenckii s.l. from 1 of 74 soil samples from armadillos’ burrows. The soil was covered with moss and partially protected from exposure to sunshine. Armadillos have been associated with the epidemiology of important human pathogens such as Mycobacterium leprae [30], Toxoplasma gondii, and Leptospira spp. [31] and dimorphic fungi such as Paracoccidioides brasiliensis [32] and Coccidioides spp. [33]. Sporotrichosis has been reported in the nine-banded armadillo D. novemcinctus [34, 35]. Recently in Brazil, 10 cases of human sporotrichosis were reported by Alves et al. [36]. The cases were associated with trauma caused by armadillo scratches during hunting. The ecological conditions, such as plants, temperature, and moisture, contribute to an armadillo’s burrow being considered a good environment for the development of Sporothrix spp. Due to the digging and foraging behavior associated with a sporadic migratory habit, armadillos may play an important role in the biogeography of pathogenic and non-pathogenic species of the genus Sporothrix by revolving soil layers and exposing the microorganisms to the surface in different environments. Moreover, armadillo hunters commonly report traumatic injuries from scratches, which can be a route of entry for Sporothrix propagules.

Mycopathologia Fig. 3 Phylogenetic relationships of S. schenckii complex isolates inferred from CAL sequences by neighbor-joining and maximum likelihood analyses based on the Kimura 2-parameter model (K2-P ? G). The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1,000 replicates) is shown next to the branches (NJ/ML) (bootstrap support values [80 are indicated in bold). GenBank accessions numbers are indicated next to strain code

Studies on the molecular ecology of clinical Sporothrix spp. are scarce. No previous study has performed molecular characterization of soilborne Sporothrix specimens and morphologically classified as S. schenckii s.l.; thus, comparisons are fairly difficult. In light of recent taxonomic changes in the genus Sporothrix and from an eco-epidemiological point of view, the frequency with which pathogenic species in the S. schenckii complex are recovered from soil should be reassessed and its classification followed by sequencing of calmodulin [1, 5] and/or the ITS region (including ITS1 ? 5.8S ? ITS2) [4] to avoid taxonomic

mistakes. Morphologically similar non-pathogenic Sporothrix species may be isolated with pathogenic Sporothrix from the same substrate [29]. Recently, Criseo and Romeo [37], based on sequencing analysis of the D1-D2 region of the 28S rRNA gene, identified 26 environmental strains of S. schenckii s.l in commercial garden soil, which is widely used in the gardens, parks, and squares in Italy. A calmodulin-based phylogeny showed the strains evaluated by Criseo and Romeo [37] belonged to Sporothrix pallida (syn = Sporothrix albicans) [7], an environmental species that is frequently isolated from

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soil and decaying wood [38], which is non-virulent to mammals [9] and a rare agent of human sporotrichosis [4]. Zhou et al. [4] found that different ecologies are corroborated by different phylogenies; i.e., the clinical clade is phylogenetically distant from the environmental clade. A comparison of ecological habits and phylogenetic analysis of clinical species in the S. schenckii complex reveal they are flanked by environmental species of Sporothrix and Ophiostoma, which are mainly associated with bark beetles and Protea or plant pathogens such as Ophiostoma ulmi and O. novoulmi (Dutch elm disease) [11, 39, 40]. Basal Sporothrix species directly isolated from soil include Sporothrix inflata, S. humicola, S. pallida, S. mexicana, S. dimorphospora, S. brunneoviolacea, and Ophiostoma species such as O. stenoceras and O. bragantinum. The ecological origins of soilborne species in the Ophiostoma–Sporothrix complex are well discussed by de Meyer et al. [38] and Madrid et al. [41]. A pathogenic strain of S. globosa was recovered and characterized from soil in Las Dichas, Chile [42], and was associated with a lymphocutaneous case of sporotrichosis. According to the phylogenetic relationship established in the present study, our soilborne strain belongs to the S. schenckii Clade IIa, a homogenous Clade that includes most of the South American strains (Argentina, Bolivia, Colombia, and Peru), adjacent to the clade harboring the North American clinical isolates, including the type strain CBS 359.36 from Maryland, USA. On the other hand, Clade IIb appears to be geographically restricted to Latin America and is composed mainly of isolates from Peru and Argentina [4, 5]. The minimum genome size of isolate Ss61 was estimated in 37.4 Mb by Sasaki et al. [43], comprising at least seven chromosomal bands (7.0, 6.7, 6.5, 5.8, 5.4, 3.4, and 2.6 Mb), which deviates from clinical S. schenckii isolates that usually presents 5–6 chromosomal bands and has an average genome size of 29.3 Mb. In addition, gene synteny evidenced by radiolabeling probes and Southern blot hybridization revealed punctual differences in isolate Ss61, what may be related to different genomic organization [43]. Antifungal resistance is a potential problem for the treatment of clinical cases of sporotrichosis. The antifungal susceptibility testing showed that our isolate was tolerant to all drugs evaluated with the exception of itraconazole, which is currently considered the best drug for treating sporotrichosis. This finding is in

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agreement with previous results reported for this species [10] as well as for the clinical isolates from armadillo hunters in southern Brazil [36]. The high MICs found in this study for different drugs highlight the need to identify the species in the S. schenckii complex to choose the best treatment. The isolation of S. schenckii s. str. from the burrows of Dasypus novemcinctus supports that armadillos may play an important role in the biological cycle of this pathogen and its eco-epidemiology during digging and foraging behavior. This directly affects the patterns of distributions of some species; in addition, it is closely related to different vectors in different geographic areas and the introduction of new routes of infection and risk factors for acquiring sporotrichosis. Further studies should be conducted to elucidate the evolutionary origin and other niches of species in the S. schenckii complex. Acknowledgments A.M.R is a fellow and acknowledges the financial support of the Fundac¸a˜o de Amparo a` Pesquisa do Estado de Sa˜o Paulo (FAPESP-2011/07350-1). E.B and S.M.G.B (FAPESP-1998/03695-8), and Z.P.C acknowledge the financial support of FAPESP (2009/54024-2). The authors also thanks to Professor Mario Rubens Guimara˜es Montenegro (in memorian) for histopathology analysis and helpful discussions.

References 1. Rodrigues AM, de Hoog S, de Camargo ZP. Emergence of pathogenicity in the Sporothrix schenckii complex. Med Mycol. 2013;51:405–12. 2. Rodrigues AM, de Melo Teixeira M, de Hoog GS, Schubach TMP, Pereira SA, Fernandes GF, et al. Phylogenetic analysis reveals a high prevalence of Sporothrix brasiliensis in feline sporotrichosis outbreaks. PLoS Negl Trop Dis 2013;7(6):e2281. 3. Marimon R, Gene´ J, Cano J, Trilles L, Dos Santos Laze´ra M, Guarro J. Molecular phylogeny of Sporothrix schenckii. J Clin Microbiol. 2006;44:3251–6. 4. Zhou X, Rodrigues AM, Feng P, Hoog GS. Global ITS diversity in the Sporothrix schenckii complex. Fungal Divers. 2013;. doi:10.1007/s13225-013-0220-2. 5. Marimon R, Cano J, Gene´ J, Sutton DA, Kawasaki M, Guarro J. Sporothrix brasiliensis, S. globosa, and S. mexicana, three new Sporothrix species of clinical interest. J Clin Microbiol. 2007;45:3198–206. 6. Marimon R, Gene´ J, Cano J, Guarro J. Sporothrix luriei: a rare fungus from clinical origin. Med Mycol. 2008; 46:621–5. 7. Romeo O, Scordino F, Criseo G. New insight into molecular phylogeny and epidemiology of Sporothrix schenckii species complex based on calmodulin-encoding gene analysis of Italian isolates. Mycopathologia. 2011;172:179–86.

Mycopathologia 8. Fernandes GF, dos Santos PO, Rodrigues AM, Sasaki AA, Burger E, de Camargo ZP. Characterization of virulence profile, protein secretion and immunogenicity of different Sporothrix schenckii sensu stricto isolates compared with S. globosa and S. brasiliensis species. Virulence. 2013;4:241–9. 9. Arrillaga-Moncrieff I, Capilla J, Mayayo E, Marimon R, Marine´ M, Gene´ J, et al. Different virulence levels of the species of Sporothrix in a murine model. Clin Microbiol Infect. 2009;15:651–5. 10. Marimon R, Serena C, Gene´ J, Cano J, Guarro J. In vitro antifungal susceptibilities of five species of Sporothrix. Antimicrob Agents Chemother. 2008;52:732–4. 11. Zipfel RD, de Beer ZW, Jacobs K, Wingfield BD, Wingfield MJ. Multi-gene phylogenies define Ceratocystiopsis and Grosmannia distinct from Ophiostoma. Stud Mycol. 2006; 55:75–97. 12. Costa EO, Diniz LS, Netto CF, Arruda C, Dagli ML. Epidemiological study of sporotrichosis and histoplasmosis in captive Latin American wild mammals, Sa˜o Paulo. Braz Mycopathol. 1994;125:19–22. 13. Rodrigues MT, de Resende MA. Epidemiologic skin test survey of sensitivity to paracoccidioidin, histoplasmin and sporotrichin among gold mine workers of Morro Velho mining. Braz Mycopathol. 1996;135:89–98. 14. Barrozo LV, Benard G, Silva MES, Bagagli E, Marques SA, Mendes RP. First description of a cluster of acute/subacute paracoccidioidomycosis cases and its association with a climatic anomaly. PLoS Negl Trop Dis. 2010;4(3):e643. 15. O’Donnell K, Nirenberg H, Aoki T, Cigelnik E. A multigene phylogeny of the Gibberella fujikuroi species complex: detection of additional phylogenetically distinct species. Mycoscience. 2000;41:61–78. 16. Liu K, Warnow TJ, Holder MT, Nelesen SM, Yu J, Stamatakis AP, et al. SATe´-II: very fast and accurate simultaneous estimation of multiple sequence alignments and phylogenetic trees. Syst Biol. 2012;61:90–106. 17. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. 2011;28:2731–9. 18. Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol. 1980;16:111–20. 19. de Vienne DM, Giraud T, Martin OC. A congruence index for testing topological similarity between trees. Bioinformatics. 2007;23:3119–24. 20. Rogers AL, Beneke ES. Human pathogenic fungi recovered from Brazilian soil. Mycopathol Mycol Appl. 1964;22:15–20. 21. Silva ME. Isolation of Histoplasma capsulatum from soil in an endemic area of kala-azar, in Bahia. Braz Fundac¸a˜o Gonc¸alo Moniz. 1956;10:15. 22. Howard DH, Orr GF. Comparison of strains of Sporotrichum schenckii isolated from nature. J Bacteriol. 1963; 85:816–21. 23. Vismer HF, Hull PR. Prevalence, epidemiology and geographical distribution of Sporothrix schenckii infections in Gauteng. South Afr Mycopathol. 1997;137:137–43. 24. Ishizaki H, Kawasaki M, Mochizuki T, Jin XZ, Kagawa S. Environmental isolates of Sporothrix schenckii in China. Nihon Ishinkin Gakkai Zasshi. 2002;43:257–60.

25. Mendoza M, Diaz E, Alvarado P, Romero E. Bastardo de Albornoz MC. Isolation of Sporothrix schenckii from environmental samples in Venezuela. Rev Iberoam Micol. 2007;24:317–9. 26. Mehta KIS, Sharma NL, Kanga AK, Mahajan VK, Ranjan N. Isolation of Sporothrix schenckii from the environmental sources of cutaneous sporotrichosis patients in Himachal Pradesh, India: results of a pilot study. Mycoses. 2007;50: 496–501. 27. Sanchez-Aleman MA, Araiza J, Bonifaz A. Isolation and characterization of wild Sporothrix schenkii strains and investigation of sporototrichin reactors. Gac Medics de Mexico. 2004;140:507–12. 28. Dixon DM, Salkin IF, Duncan RA, Hurd NJ, Haines JH, Kemna ME, et al. Isolation and characterization of Sporothrix schenckii from clinical and environmental sources associated with the largest U.S. epidemic of sporotrichosis. J Clin Microbiol. 1991;29:1106–13. 29. Mackinnon JE, Conti-Dı´az IA, Gezuele E, Civila E, Da Luz S. Isolation of Sporothrix schenckii from nature and considerations on its pathogenicity and ecology. Med Mycol. 1969;7:38–45. 30. Truman RW, Shannon EJ, Hagstad HV, Hugh-Jones ME, Wolff A, Hastings RC. Evaluation of the origin of Mycobacterium leprae infections in the wild armadillo, Dasypus novemcinctus. Am J Trop Med Hyg. 1986;35:588–93. 31. da Silva RC, Zetun CB, Bosco Sde M, Bagagli E, Rosa PS, Langoni H. Toxoplasma gondii and Leptospira spp. infection in free-ranging armadillos. Vet Parasitol. 2008;157:291–3. 32. Bagagli E, Sano A, Coelho KI, Alquati S, Miyaji M, de Camargo ZP, et al. Isolation of Paracoccidioides brasiliensis from armadillos (Dasypus noveminctus) captured in an endemic area of paracoccidioidomycosis. Am J Trop Med Hyg. 1998;58:505–12. 33. Costa FAMd, Reis RC, Benevides F, Tome´ GdS, Holanda MA. Pulmonary coccidioidomycosis in a armadillo hunter. J Pneumologia. 2001;27:275–8. 34. Wenker CJ, Kaufman L, Bacciarini LN, Robert N. Sporotrichosis in a nine-banded armadillo (Dasypus novemcinctus). J Zoo Wildl Med. 1998;29:474–8. 35. Kaplan W, Broderson JR, Pacific JN. Spontaneous systemic sporotrichosis in nine-banded armadillos (Dasypus novemcinctus). Sabouraudia. 1982;20:289–94. 36. Alves SH, Boettcher CS, Oliveira DC, Tronco-Alves GR, Sgaria MA, Thadeu P, et al. Sporothrix schenckii associated with armadillo hunting in Southern Brazil: epidemiological and antifungal susceptibility profiles. Rev Soc Bras Med Trop. 2010;43:523–5. 37. Criseo G, Romeo O. Ribosomal DNA sequencing and phylogenetic analysis of environmental Sporothrix schenckii strains: comparison with clinical isolates. Mycopathologia. 2010;169:351–8. 38. de Meyer EM, de Beer ZW, Summerbell RC, Moharram AM, de Hoog GS, Vismer HF, et al. Taxonomy and phylogeny of new wood- and soil-inhabiting Sporothrix species in the Ophiostoma stenoceras–Sporothrix schenckii complex. Mycologia. 2008;100:647–61. 39. Roets F, Wingfield BD, de Beer ZW, Wingfield MJ, Dreyer LL. Two new Ophiostoma species from Protea caffra in Zambia. Persoonia-Mol Phylogeny Evol Fungi. 2010;24:18–28.

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Mycopathologia 40. Temple B, Pines PA, Hintz WE. A nine-year genetic survey of the causal agent of Dutch elm disease, Ophiostoma novoulmi in Winnipeg. Can Mycol Res. 2006;110:594–600. 41. Madrid H, Gene´ J, Cano J, Silvera C, Guarro J. Sporothrix brunneoviolacea and Sporothrix dimorphospora, two new members of the Ophiostoma stenoceras–Sporothrix schenckii complex. Mycologia. 2010;102:1193–203.

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42. Cruz R, Vieille P, Oschilewski D. Sporothrix globosa isolation related to a case of lymphocutaneous sporotrichosis. Rev Chil Infectol. 2012;29:401–5. 43. Sasaki AA, Fernandes GF, Rodrigues AM, Lima FM, Marini MM, Feitosa LS, et al. Chromosomal polymorphisms in the Sporothrix schenckii complex. PLoS ONE. doi:10.1371/ journal.pone.0086819.

Sporothrix schenckii sensu stricto isolated from soil in an armadillo's burrow.

Sporotrichosis is a polymorphic disease of man and animals caused by traumatic implantation of propagules into the skin and subcutaneous tissue. Patho...
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