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

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

Eighty Years of Mycopathologia: A Retrospective Analysis of Progress Made in Understanding Human and Animal Fungal Pathogens

Mycopathologia was founded in 1938 to \u27diffuse the understanding of fungal diseases in man and animals among mycologists.\u27 This was an important mission considering that pathogenic fungi for humans and animals represent a tiny minority of the estimated 1.5-5 million fungal inhabitants on Earth. These pathogens have diverged from the usual saprotrophic lifestyles of most fungi to colonize and infect humans and animals. Medical and veterinary mycology is the subdiscipline of microbiology that dwells into the mysteries of parasitic, fungal lifestyles. Among the oldest continuing scientific publications on the subject, Mycopathologia had its share of \u27classic papers\u27 since the first issue was published in 1938. An analysis of the eight decades of notable contributions reveals many facets of host-pathogen interactions among 183 volumes comprising about 6885 articles. We have analyzed the impact and relevance of this body of work using a combination of citation tools (Google Scholar and Scopus) since no single citation metric gives an inclusive perspective. Among the highly cited Mycopathologia publications, those on experimental mycology accounted for the major part of the articles (36%), followed by diagnostic mycology (16%), ecology and epidemiology (15%), clinical mycology (14%), taxonomy and classification (10%), and veterinary mycology (9%). The first classic publication, collecting nearly 200 citations, appeared in 1957, while two articles published in 2010 received nearly 150 citations each, which is notable for a journal covering a highly specialized field of study. An empirical analysis of the publication trends suggests continuing interests in novel diagnostics, fungal pathogenesis, review of clinical diseases especially with relevance to the laboratory scientists, taxonomy and classification of fungal pathogens, fungal infections and carriage in pets and wildlife, and changing ecology and epidemiology of fungal diseases around the globe. We anticipate that emerging and re-emerging fungal pathogens will continue to cause significant health burden in the coming decades. It remains vital that scientists and physicians continue to collaborate by learning each other\u27s language for the study of fungal diseases, and Mycopathologia will strive to be their partner in this increasingly important endeavor to its 100th anniversary in 2038 and beyond

HISTOPLASMIN SURVEY IN HIV-POSITIVE PATIENTS: RESULTS FROM AN ENDEMIC AREA IN NORTHEASTERN BRAZIL

SUMMARY Background. Disseminated histoplasmosis is common in AIDS patients with advanced immunosuppression in Ceará, Northeastern Brazil. The goal of this study was to determine the prevalence of Histoplasma infection in patients with HIV/AIDS living in Fortaleza, the capital of Ceará. Methods. Intradermal tests with histoplasmin (mycelial phase) were performed in 161 HIV patients with CD4 ≥ 350 cells/mm 3 . Evidence of recent illness was evaluated with immunodiffusion (ID) tests in 76 of these individuals. Results. A total of 11.8% of patients reacted to histoplasmin and 2.63% had ID test positive to Histoplasma. The presence of mango trees (Mangifera indica) in the patient neighborhood (OR = 2.870; 95% CI = 1.081-7.617; p = 0.040) and past activity involving soil (OR = 2.834; 95% CI = 1.045-7.687; p = 0.045) or visits to a farm (OR = 3.869; 95% CI = 1.189-12.591; p = 0.033) were significantly associated with Histoplasma infection. Conclusions. Patients with HIV living in Fortaleza have an expressive prevalence of infection with Histoplasma

CO2 laser irradiation enhances CaF2 formation and inhibits lesion progression on demineralized dental enamel—in vitro study

This study evaluated if Carbon dioxide (CO2) (lambda 10.6 mu m) laser irradiation combined with acidulated phosphate fluoride gel application (APF gel) enhances "CaF2" uptake by demineralized enamel specimens (DES) and inhibits enamel lesion progression. Thus, two studies were conducted and DES were subjected to APF gel combined or not with CO2 laser irradiation (11.3 or 20.0 J/cm(2), 0.4 or 0.7 W) performed before, during, or after APF gel application. In study 1, 165 DES were allocated to 11 groups. Fluoride as "CaF2 like material" formed on enamel was determined in 100 DES (n = 10/group), and the surface morphologies of 50 specimens were evaluated by scanning electron microscopy (SEM) before and after "CaF2" extraction. In study 2, 165 DES (11 groups, n = 15), subjected to the same treatments as in study 1, were further subjected to a pH-cycling model to simulate a high cariogenic challenge. The progression of demineralization in DES was evaluated by cross-sectional microhardness and polarized light microscopy analyses. Laser at 11.3 J/cm(2) applied during APF gel application increased "CaF2" uptake on enamel surface. Laser irradiation and APF gel alone arrested the lesion progression compared with the control (p < 0.05). Areas of melting, fusion, and cracks were observed. CO2 laser irradiation, combined with a single APF application enhanced "CaF2" uptake on enamel surface and a synergistic effect was found. However, regarding the inhibition of caries lesion progression, no synergistic effect could be demonstrated. In conclusion, the results have shown that irradiation with specific laser parameters significantly enhanced CaF2 uptake by demineralized enamel and inhibited lesion progression313539547sem informaçã

Histoplasma capsulatum synthesizes melanin-like pigments in vitro and during mammalian infection

Melanin is made by several important pathogenic fungi and has been implicated in the pathogenesis of a number of fungal infections. This study investigated whether the thermally dimorphic fungal pathogen Histoplasma capsulatum var. capsulatum produced melanin or melanin-like compounds in vitro and during infection. Growth of H. capsulatum mycelia in chemically defined minimal medium produced pigmented conidia. Growth of H. capsulatum yeast in chemically defined minimal medium with l-3,4-dihydroxyphenylalanine (DOPA) or (-)-epinephrine produced pigmented cells. Treatment of the pigmented cells with proteolytic enzymes, denaturant, and hot concentrated acid yielded dark particles that were similar in size and shape to their respective propagules. Melanin-binding monoclonal antibodies (MAb) labeled pigmented conidia, yeast, and the isolated particles as determined by immunofluorescence microscopy. Electron spin resonance spectroscopy revealed that pigmented yeast cells and particles derived from pigmented cells were stable free radicals consistent with their identification as melanins. Tissues from mice infected with H. capsulatum and from biopsy specimens from a patient with histoplasmosis contained fungal cells that were labeled by melanin-binding MAb. Digestion of infected mouse tissues yielded dark particles that reacted with the melanin-binding MAb and were similar in appearance to H. capsulatum yeast cells. Additionally, sera from infected mice contained antibodies that bound melanin particles. Phenoloxidase activity capable of synthesizing melanin from L-DOPA was detected in cytoplasmic yeast cell extracts. These findings indicate that H. capsulatum conidia and yeast can produce melanin or melanin-like compounds in vitro and that yeast cells can synthesize pigment in vivo. Since melanin is an important virulence factor in other pathogenic fungi, this pigment may have a similar role to play in the pathogenesis of histoplasmosis

SHORT REPORT Isolation of Histoplasma capsulatum from bats in the urban area of São Paulo State, Brazil

The presence of bats in caves, attics, ceilings, and roofs is important epidemiologically as they can increase the chance of human acquisition of pathogens, including Histoplasma capsulatum. Brazilian urban areas contain many species of bats, especially insectivorous bats, that are attracted by a wide range of readily available food and shelter. From August 2003 to December 2008, we analysed 2427 bats in the São Paulo State region. Homogenates of the livers and spleens of the bats were plated on specific medium to identify animals infected with H. capsulatum. the fungus was isolated from 87 bats (3.6%). the infected bats were identified as Molossus molossus (74), Nyctinomops macrotis (10), Tadarida brasiliensis (1), Molossus rufus (1) and Eumops glaucinus (1), all insectivorous species. the data presented are a relevant contribution to the epidemiology of H. capsulatum in densely populated urban areas such as in São Paulo State, especially since histoplasmosis is not included in the mandatory disease notification system.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Univ São Paulo, Inst Ciencias Biomed, Dept Microbiol, BR-05508000 São Paulo, BrazilZoonosis Control Ctr Municipal São Paulo, São Paulo, BrazilInst Oswaldo Cruz Fdn, Immunodiagnost Sect, Lab Mycol, Evandro Chagas Inst Clin Res, Rio de Janeiro, BrazilAlbert Einstein Coll Med, Dept Med, the Bronx, NY USAAlbert Einstein Coll Med, Dept Microbiol & Immunol, the Bronx, NY USAUniversidade Federal de São Paulo, Div Cell Biol, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilUniv São Paulo, Med Mycol Lab, LIM53, HCFMUSP, BR-05508000 São Paulo, BrazilUniv São Paulo, Inst Trop Med, BR-05508000 São Paulo, BrazilUniversidade Federal de São Paulo, Div Cell Biol, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilFAPESP: 06/58210-7FAPESP: 07/07588-2Web of Scienc

International evaluation of CLSI MIC/MEC distributions for definition of epidemiological cutoff values (ECVs) for species of Sporothrix identified by molecular methods

VCU Med Ctr, Richmond, VA USAUniv Fed Rural Rio de Janeiro, Seropedica, BrazilInst Nacl Infectol, Fundação Oswaldo Cruz Fiocruz, Rio De Janeiro, BrazilUniv Fed Ceara, Specialized Med Mycol Ctr, Fortaleza, Ceara, BrazilPostgrad Inst Med Educ, Chandigarh, IndiaUniv Delhi, Vallabhbhai Patel Chest Inst, Delhi, IndiaCanisius Wilhelmina Hosp, Ctr Expertise Mycol, Nijmegen, NetherlandsInst Nacl Enfermedades Infecciosas Dr C, Dept Micol, Buenos Aires, DF, ArgentinaUniv Autonoma Nuevo Leon, Monterrey, MexicoNatl Inst Communicable Dis, Johannesburg, South AfricaUniv Witwatersra, Johannesburg, South AfricaPubl Hlth England, Mycol Reference Lab, Bristol, Avon, EnglandSA Pathol, Natl Mycol Reference Ctr, Adelaide, SA, AustraliaInst Nacl Infectol Evandro Chagas, Fundação Oswaldo Cruz Fiocruz, Rio De Janeiro, BrazilUniv Fed São Paulo, São Paulo, BrazilInst Biofis, São Paulo, BrazilUniv Fed Rio de Janeiro, Inst Biofis, Rio De Janeiro, BrazilRio Claro Labs, Inst Adolfo Lutz, São Paulo, BrazilHosp Clin Dr M Quintela, Dept Lab Clin, Montevideo, UruguayHosp Gen Mexico City, Mexico City, DF, MexicoUniv Rovira & Virgili, Mycol Unit, Sch Med, Reus, SpainInst Nacl Higiene Rafael Rangel, Caracas, VenezuelaUniv Antioquia, Grp Invest Dermatol, Medellin, ColombiaUniv Adelaide, Adelaide, SA, AustraliaUniv Fed São Paulo, São Paulo, BrazilWeb of Scienc