Sporotrichosis caused by sporothrix mexicana, Portugal

Sporotrichosis is a subcutaneous fungal infection present worldwide that is caused by traumatic inoculation or inhalation of spores of the dimorphic fungus Sporothrix schenckii complex (1-3). However, molecular studies have shown that the S. schenckii complex constitutes several cryptic infectious species (i.e., S. albicans, S. brasiliensis, S. globosa, S. luriei, S. mexicana, and S. schenckii). Marimon et al. (4) demonstrated 3 major clades grouped into 6 putative phylogenetic species. The natural habitats of these species are soil and plants. The species showed distinct pathologic behavior, antifungal responses, and phenotypes, which suggests that optimal clinical treatment may depend on the taxon involved in the sporotrichosis (1). Human infections have been reported primarily from the Americas, including Latin America (3,5). Asia (e.g., Malaysia, India, Japan), Africa, and Australia are also regions where infections are endemic (6). Although infections are rare in Europe, a case of human infection (7) and a case of an animal infection (8) have been described in southern Italy. We report a case of human sporotrichosis in which S. mexicana was isolated from a patient in Portugal.(undefined

Development and optimization of a new MALDI-TOF protocol for identification of the Sporothrix species complex

Accurate species identification of the Sporothrix schenckii complex is essential, since identification based only on phenotypic characteristics is often inconclusive due to phenotypic variability within the species. We used matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for species identification of 70 environmental and clinical isolates of the Sporothrix complex. A reference database was established for MALDI-TOF MS-based species identification according to minor adjustments in the manufacturer's guidelines. The MALDI-TOF MS clearly distinguished strains of Sporothrix brasiliensis, Sporothrix globosa, Sporothrix mexicana, S. schenckii, Sporothrix luriei and Sporothrix pallida, enabling identification of all isolates at the species level, as confirmed by partial calmodulin gene sequence analyses. The present methodology is simple, reliable, rapid and highly suitable for routine identification in clinical mycology laboratories and culture collections, particularly for updating and reclassifying of deposited Sporothrix isolates.The authors wish to thank the following international researchers for generously contributing strains to this study: Conchita Torrielo (EH194, EH252, EH253); Myrtha Arango (04015, 11029, 010221, 10036, 03017, 03022, 12013, 03003, 14879); and Masako Kawasaki (KMU975). Financial support was provided by FAPERJ/Rio de Janeiro, Brazil (grant proc. E-26/110.619/2012) and PAPES VI-Fiocruz/CNPq (Proc. 407693/2012-2) R. M. Z-O. is supported in part by CNPq 304976/2013-0 and FAPERJ E-26/103.157/2011. M. M. E. O. was supported by a grant from CAPES 2445/11-5 and PNPD/CAPES-Fiocruz/Pesquisa Clinica em Doencas Infecciosas. M. M. E. O., C. S. and N. L. thank the FCT Strategic Project PEst-OE/EQB/LA0023/2013 and BioHealth-Biotechnology and Bioengineering approaches to improve health quality, Ref. NORTE-07-0124-FEDER-000027" co-funded by the Programa Operacional Regional do Norte (ON.2 - O Novo Norte), QREN, FEDER. Automated sequencing was done using the genomic platform/DNA sequencing platform at the Fundacao Oswaldo Cruz-PDTIS/FIOCRUZ (RPT01A), Brazil

Crosstalk between Medulloblastoma Cells and Endothelium Triggers a Strong Chemotactic Signal Recruiting T Lymphocytes to the Tumor Microenvironment

Cancer cells can live and grow if they succeed in creating a favorable niche that often includes elements from the immune system. While T lymphocytes play an important role in the host response to tumor growth, the mechanism of their trafficking to the tumor remains poorly understood. We show here that T lymphocytes consistently infiltrate the primary brain cancer, medulloblastoma. We demonstrate, both in vitro and in vivo, that these T lymphocytes are attracted to tumor deposits only after the tumor cells have interacted with tumor vascular endothelium. Macrophage Migration Inhibitory Factor (MIF)” is the key chemokine molecule secreted by tumor cells which induces the tumor vascular endothelial cells to secrete the potent T lymphocyte attractant “Regulated upon Activation, Normal T-cell Expressed, and Secreted (RANTES).” This in turn creates a chemotactic gradient for RANTES-receptor bearing T lymphocytes. Manipulation of this pathway could have important therapeutic implications

Multicenter, International Study of MIC/MEC Distributions for Definition of Epidemiological Cutoff Values for Sporothrix Species Identified by Molecular Methods

ABSTRACT Clinical and Laboratory Standards Institute (CLSI) conditions for testing the susceptibilities of pathogenic Sporothrix species to antifungal agents are based on a collaborative study that evaluated five clinically relevant isolates of Sporothrix schenckii sensu lato and some antifungal agents. With the advent of molecular identification, there are two basic needs: to confirm the suitability of these testing conditions for all agents and Sporothrix species and to establish species-specific epidemiologic cutoff values (ECVs) or breakpoints (BPs) for the species. We collected available CLSI MICs/minimal effective concentrations (MECs) of amphotericin B, five triazoles, terbinafine, flucytosine, and caspofungin for 301 Sporothrix schenckii sensu stricto, 486 S. brasiliensis, 75S. globosa, and 13 S. mexicana molecularly identified isolates. Data were obtained in 17 independent laboratories (Australia, Europe, India, South Africa, and South and North America) using conidial inoculum suspensions and 48 to 72 h of incubation at 35°C. Sufficient and suitable data (modal MICs within 2-fold concentrations) allowed the proposal of the following ECVs for S. schenckii and S. brasiliensis, respectively: amphotericin B, 4 and 4 g/ml; itraconazole, 2 and 2 g/ml; posaconazole, 2 and 2 g/ml; and voriconazole, 64 and 32 g/ml. Ketoconazole and terbinafine ECVs for S. brasiliensis were 2 and 0.12 g/ml, respectively. Insufficient or unsuitable data precluded the calculation of ketoconazole and terbinafine (or any other antifungal agent) ECVs for S. schenckii, as well as ECVs for S. globosa and S. mexicana. These ECVs could aid the clinician in identifying potentially resistant isolates (non-wild type) less likely to respond to therapy

Molecular detection of Paracoccidioides brasiliensis in road-killed wild animals

Paracoccidioides brasiliensis infections have been little studied in wild and/or domestic animals, which may represent an important indicator of the presence of the pathogen in nature. Road-killed wild animals have been used for surveillance of vectors of zoonotic pathogens and may offer new opportunities for eco-epidemiological studies of paracoccidiodomycosis (PCM). The presence of P. brasiliensis infection was evaluated by Nested-PCR in tissue samples collected from 19 road-killed animals; 3 Cavia aperea (guinea pig), 5 Cerdocyon thous (crab-eating-fox), 1 Dasypus novemcinctus (nine-banded armadillo), 1 Dasypus septemcinctus (seven-banded armadillo), 2 Didelphis albiventris (white-eared opossum), 1 Eira barbara (tayra), 2 Gallictis vittata (grison), 2 Procyon cancrivorus (raccoon) and 2 Sphiggurus spinosus (porcupine). Specific P. brasiliensis amplicons were detected in (a) several organs of the two armadillos and one guinea pig, (b) the lung and liver of the porcupine, and (c) the lungs of raccoons and grisons. P. brasiliensis infection in wild animals from endemic areas might be more common than initially postulated. Molecular techniques can be used for detecting new hosts and mapping `hot spot` areas of PCM

Molecular detection of Paracoccidioides brasiliensis in road-killed wild animals

Paracoccidioides brasiliensis infections have been little studied in wild and/or domestic animals, which may represent an important indicator of the presence of the pathogen in nature. Road-killed wild animals have been used for surveillance of vectors of zoonotic pathogens and may offer new opportunities for eco-epidemiological studies of paracoccidiodomycosis (PCM). The presence of P. brasiliensis infection was evaluated by Nested-PCR in tissue samples collected from 19 road-killed animals; 3 Cavia aperea (guinea pig), 5 Cerdocyon thous (crab-eating-fox), 1 Dasypus novemcinctus (nine-banded armadillo), 1 Dasypus septemcinctus (seven-banded armadillo), 2 Didelphis albiventris (white-eared opossum), 1 Eira barbara (tayra), 2 Gallictis vittata (grison), 2 Procyon cancrivorus (raccoon) and 2 Sphiggurus spinosus (porcupine). Specific P. brasiliensis amplicons were detected in (a) several organs of the two armadillos and one guinea pig, (b) the lung and liver of the porcupine, and (c) the lungs of raccoons and grisons. P. brasiliensis infection in wild animals from endemic areas might be more common than initially postulated. Molecular techniques can be used for detecting new hosts and mapping `hot spot` areas of PCM