Biochemical Characterization of an In-House Coccidioides Antigen: Perspectives for the Immunodiagnosis of Coccidioidomycosis

The objective of this study was to evaluate the reactivity of an in-house antigen, extracted from a strain of C. posadasii isolated in northeastern Brazil, by radial immunodiffusion and Western blotting, as well as to establish its biochemical characterization. the protein antigen was initially extracted with the use of solid ammonium sulfate and characterized by 1-D electrophoresis. Subsequently, it was tested by means of double radial immunodiffusion and Western blotting. A positive reaction was observed against the antigen by both immunodiagnostic techniques tested on sera from patients suffering from coccidioidomycosis. Besides this, two immunoreactive protein bands were observed and were revealed to be a beta-glucosidase and a glutamine synthetase after sequencing of the respective N-terminal regions. Our in-house Coccidioides antigen can be promising as a quick and low-cost diagnostic tool without the risk of direct manipulation of the microorganism.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordination for the Improvement of Higher Education PersonnelUniv Fed Ceara, Specialized Med Mycol Ctr, Sch Med, BR-60430270 Fortaleza, Ceara, BrazilUniv Estadual Ceara, Postgrad Program Vet Sci, Sch Vet Med, BR-60740000 Fortaleza, Ceara, BrazilUniv Fed Ceara, Dept Biochem & Mol Biol, BR-60455760 Fortaleza, Ceara, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, BR-04021001 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, BR-04021001 São Paulo, BrazilCNPq: 302574/2009-3CNPq: 306637/2010-3Coordination for the Improvement of Higher Education Personnel: CAPES/PNPD2103/2009Web of Scienc

Antiretroviral drugs saquinavir and ritonavir reduce inhibitory concentration values of itraconazole against Histoplasma capsulatum strains in vitro

Recent studies have shown that some drugs that are not routinely used to treat fungal infections have antifungal activity, such as protease inhibitor antiretroviral drugs. This study investigated the in vitro susceptibility of Histoplasma capsulatum var. capsulatum to saquinavir and ritonavir, and its combination with the antifungal itraconazole. The susceptibility assay was performed according to Clinical and Laboratory Standards Institute guidelines. All strains were inhibited by the protease inhibitor antiretroviral drugs. Saquinavir showed minimum inhibitory concentrations ranging from 0.125 to 1 mu g mL(-1) for both phases, and ritonavir presented minimum inhibitory concentrations ranging from 0.0312 to 4 mu g mL(-1) and from 0.0625 to 1 mu g mL(-1) for filamentous and yeast phase, respectively. Concerning the anti fungal itraconazole, the minimum inhibitory concentration values ranged from 0.0019 to 0.125 mu g mL(-1) and from 0.0039 to 0.0312 mu g mL(-1) for the filamentous and yeast phase, respectively. The combination of saquinavir or ritonavir with itraconazole was synergistic against H. capsulatum, with a significant reduction in the minimum inhibitory concentrations of both drugs against the strains (p < 0.05). These data show an important in vitro synergy between protease inhibitors and itraconazole against the fungus H. capsulatum. (C) 2016 Published by Elsevier Editora Ltda.CNPqCAPESUniv Fed Ceara, Postgrad Program Med Microbiol, Specialized Med Mycol Ctr, Fortaleza, CE, BrazilUniv Fed Ceara, Postgrad Program Med Sci, Fortaleza, CE, BrazilUniv Estadual Ceara UECE, Postgrad Program Vet Sci, Fortaleza, CE, BrazilUniv Fed Ceara, Dept Stat & Appl Math, Fortaleza, CE, BrazilUniv Fed Sao Paulo UNIFESP, Dept Microbiol Immunol & Parasitol, Sao Paulo, SP, BrazilHosp Sao Jose, Fortaleza, CE, BrazilUniv Fed Sao Paulo UNIFESP, Dept Microbiol Immunol & Parasitol, Sao Paulo, SP, BrazilCNPq: 303396/2014-8CNPq: 552161/2011-0CAPES: AE1 - 0052-000630100/11Web of Scienc

Coccidioidomycosis and Histoplasmosis in Equines: An Overview to Support the Accurate Diagnosis

Fungal infections of the respiratory tract of horses are not as frequent as those of bacterial and viral origin, often leading to worsening of clinical conditions due to misdiagnosis and incorrect treatment. Coccidioidomycosis and histoplasmosis are systemic mycoses caused by the dimorphic fungi Coccidioides spp. and Histoplasma capsulatum, respectively, which affect humans and a variety of other animals, including equines. These systemic mycoses of chronic and progressive nature can exhibit clinical manifestations similar to other microbial infections. Thus, this article broadly discusses the epidemiology, etiology, virulence, pathogenesis, clinical presentation, treatment, and diagnostic strategies of coccidioidomycosis and histoplasmosis, to support accurate diagnosis. (C) 2016 Elsevier Inc. All rights reserved.National Scientific and Technological Research CouncilBrazil (CNPq)Univ Fed Ceara, Postgrad Program Med Microbiol, Specialized Med Mycol Ctr, Dept Pathol & Legal Med,Sch Med, BR-60430275 Fortaleza, Ceara, BrazilUniv Estadual Ceara, Postgrad Program Vet Sci, Sch Vet, Fortaleza, Ceara, BrazilUniv Fed Sao Paulo, Dept Microbiol Immunol & Parasitol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Microbiol Immunol & Parasitol, Sao Paulo, BrazilCNPq: 445670/2014-2CNPq: 552161/2011-0Web of Scienc

In vitro susceptibility of antifungal drugs against Sporothrix brasiliensis recovered from cats with sporotrichosis in Brazil

Sporotrichosis is an important subcutaneous mycosis of humans and animals. Classically, the disease is acquired upon traumatic inoculation of Sporothrix propagules from contaminated soil and plant debris. In addition, the direct horizontal transmission of Sporothrix among animals and the resulting zoonotic infection in humans highlight an alternative and efficient rout of transmission through biting and scratching. Sporothrix brasiliensis is themost virulent species of the Sporothrix schenckii complex and is responsible for the long-lasting outbreak of feline sporotrichosis in Brazil. However, antifungal susceptibility data of animal-borne isolates is scarce. Therefore, this study evaluated the in vitro activity of amphotericin B, caspofungin, itraconazole, voriconazole, fluconazole, and ketoconazole against animal-borne isolates of S. brasiliensis. The susceptibility tests were performed through broth microdilution (M38-A2). The results show the relevant activity of itraconazole, amphotericin B, and ketoconazole against S. brasiliensis, with the following MIC ranges: 0.125-2, 0.125-4 and 0.0312-2 mu g/ml, respectively. Caspofungin was moderately effective, displaying higher variation in MIC values (0.25-64 mu g/ml). Voriconazole (2-64 mu g/ml) and fluconazole (62.5-500 mu g/ml) showed low activity against S. brasiliensis strains. This study contributed to the characterization of the in vitro antifungal susceptibility of strains of S. brasiliensis recovered from cats with sporotrichosis, which have recently been considered the main source of human infections.Sao Paulo Research Foundation (FAPESP)National Council for Scientific and Technological Development (CNPq 304779/20113)Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)Programa Estrategico de Apoio a Pesquisa em Saude VI - Fiocruz/CNPqFundacao Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ)Univ Fed Ceara, Specialized Med Mycol Ctr, Postgrad Program Med Microbiol, Fortaleza, Ceara, BrazilUniv Fed Sao Paulo, Dept Microbiol Immunol & Parasitol, Cellular Biol Div, Sao Paulo, BrazilFundacao Oswaldo Cruz, Evandro Chagas Natl Inst Infect Dis, Rio De Janeiro, BrazilUniv Fed Sao Paulo, Dept Microbiol Immunol & Parasitol, Cellular Biol Div, Sao Paulo, BrazilFAPESP: 2011/07350-1FAPESP: 2009/54024-2CNPq: 304779/20113CAPES: AE1-0052-000650100/11Fiocruz/CNPq: 407771/2013-3FAPERJ: E-26/102.255/2013Web of Scienc

Easy Storage Strategies for Sporothrix spp. Strains

The present study evaluated the maintenance of Sporothrix spp. (6 Sporothrix brasiliensis; 6 S. schenckii; 5 S. mexicana, and 3 S. globosa) in saline at 4 degrees C, and in 10% glycerol plus either 10% lactose or 10% sucrose, at -20 degrees C and -80 degrees C. Viability was assessed after 3, 6, and 9 months of storage, through the recovery of strains on potato dextrose agar and analysis of macro- and micromorphological features. Conidium quantification was performed before and after storage, at 3, 6 and 9 months. 100% viability was observed, regardless of storage conditions or time period. Storage at 4 degrees C and at -20 degrees C did not alter the number of conidia, but lower conidium counts were observed at -80 degrees C. This study shows that the combination of glycerol with lactose or sucrose is effective to maintain Sporothrix spp. at freezing temperatures.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Fed Ceara, Grad Program Med Microbiol, Specialized Med Mycol Ctr, Fortaleza, Ceara, BrazilUniv Fed Ceara, Sch Med, Christus Coll, UNICHRISTUS, Fortaleza, Ceara, BrazilUniv Fed Ceara, Dept Stat & Appl Math, Fortaleza, Ceara, BrazilUniv Fed Ceara, Grad Program Med Sci, Fortaleza, Ceara, BrazilUniv Estadual Ceara, Grad Program Vet Sci, Fortaleza, Ceara, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilCNPq: PROTAX 562296/2010-7CNPq: 304779/2011-3CNPq: 504189/2012-3Web of Scienc