Candidemia in a Brazilian hospital: the importance of Candida parapsilosis

The aim of this study was to perform a retrospective analysis of cases of candidemia in a Brazilian hospital in the city of Fortaleza, Ceará. A total of 50 blood cultures were analyzed from 40 candidemic patients. The mycological diagnosis was based on the phenotypical analysis and the patients' data were recorded in appropriate files. The most frequent species were Candida parapsilosis (n = 18), followed by C. albicans (n = 14), C. tropicalis (n = 8), C. guillermondii (n = 6), C. glabrata (n = 2), and Candida spp. (n = 2). A detailed descriptive study was undertaken with 21 patients whose medical records were complete. The candidemia episodes occurred in eight male patients and 13 female patients. The most representative risk factors implicated in candidemia were prior antibiotic therapy, central venous catheters, parenteral nutrition, gastric probes and mechanical ventilation. Death occurred in 13 of the 21-candidemic patients. This study demonstrated the emergence of candidemia caused by C. parapsilosis in a Brazilian hospital in the city of Fortaleza, Ceará.O presente estudo objetivou desenvolver uma análise retrospectiva de casos de candidemia em hospital brasileiro na cidade de Fortaleza, Ceará. Um total de 50 hemoculturas foram analisadas de 40 pacientes com quadros de candidemia. O diagnóstico micológico foi baseado na análise morfológica e bioquímica e os dados dos pacientes foram coletados das histórias clínicas. As espécies mais freqüentes foram Candida parapsilosis (n = 18), seguida por C. albicans (n = 14), C. tropicalis (n = 8), C. guillermondii (n = 6), C. glabrata (n = 2) e Candida spp. (n = 2). Um estudo descritivo foi realizado com apenas 21 pacientes os quais possuíam dados clínicos completos. Os episódios de candidemia aconteceram em oito pacientes do sexo masculino e 13 do feminino. Os fatores de risco implicados em candidemia foram antibioticoterapia prévia, uso de cateter venoso central, nutrição parenteral, sondagem gástrica e ventilação mecânica. A morte aconteceu em 13 dos 21 pacientes com candidemia. Este estudo demonstrou a emergência de candidemia causada por C. parapsilosis em um hospital brasileiro na cidade de Fortaleza, Ceará

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

Cotrimoxazole enhances the in vitro susceptibility of Coccidioides posadasii to antifungals

The aim of the present study was to evaluate the effect of cotrimoxazole on the in vitro susceptibility of Coccidioides posadasii strains to antifungals. A total of 18 strains of C. posadasii isolated in Brazil were evaluated in this study. The assays were performed in accordance with the Clinical and Laboratory Standards Institute guidelines and the combinations were tested using the checkerboard method. The minimum inhibitory concentrations were reduced by 11, 2.4, 4.3 and 3.5 times for amphotericin B, itraconazole, fluconazole and voriconazole, respectively. Moreover, it was seen that cotrimoxazole itself inhibited C. posadasii strains in vitro. The impairment of folic acid synthesis may be a potential antifungal target for C. posadasii.Universidade Federal do Ceará Centro Especializado em Micologia MédicaUniversidade Federal do Ceará Programa de Pós-Graduação em Ciências MédicasUniversidade Federal do Ceará Departamento de QuímicaUniversidade Federal do Ceará Departamento de EstatísticaUniversidade Estadual do Ceará Programa de Pós-Graduação em Ciência VeterináriaUniversidade Federal de São Paulo (UNIFESP) Departamento de Microbiologia, Imunologia e ParasitologiaUNIFESP, Depto. de Microbiologia, Imunologia e ParasitologiaSciEL

Relação altura-diâmetro para um povoamento clonal jovem de Tectona grandis Linn F. na Amazônia oriental, Brasil / Hypsometric relation for a young clonal plantation of Tectona grandis Linn F. in eastern Amazon, Brazil

A mensuração da variável altura consiste em uma operação onerosa e propícia a erros que podem superestimar ou subestimar os demais parâmetros de uma floresta. Com isso, este trabalho teve por objetivo obter um modelo hipsométrico para um plantio clonal jovem de Tectona grandis na Amazônia Oriental. Foram estabelecidas cinco parcelas permanentes, medidas aos 18 meses de idade do plantio, utilizando-se método de área fixa, com parcelas de 60 x 60 m, sob processo de amostragem aleatória simples. Com dados de 120 árvores, foram ajustados 11 modelos hipsométricos, quatro lineares e sete não lineares. As equações foram validadas pelo teste do Qui-quadrado. Para seleção foram utilizados o criterios: significância da estatística F da análise de variância (ANOVA) da regressão; coeficiente de determinação ajustado (R²aj.%); erro padrão da estimativa em percentagem (Syx%) e recalculado (Syxr%); desvio médio percentual (DMP%); e gráfico de valores observados e preditos. Nos resultados observamos correlações variando de forte à muito forte entre diâmetro e altura. Na modelagem, a equação 5 (hiperbólica), não-linear, foi a selecionada, pois apresentou melhores critérios estatísticos, com R²aj.= 92,45%, Syx% = 5,32%, DMP% = 0,002% e distribuição residual não tendenciosa e validada. Portanto, ela pode ser indicada para estimar a variável altura total dos indivíduos de T. grandis

Strategies for the knowledge of coccidioidomicose - an emergent illness in the Brazilian Northeast

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel SuperiorA coccidioidomicose à uma infecÃÃo sistÃmica causada pelos fungos dimÃrficos Coccidioides immitis e C. posadasii. A doenÃa acomete o homem e uma grande variedade de animais, sendo endÃmica no Estado do CearÃ, onde à causada pela espÃcie C. posadasii. O presente estudo buscou analisar as caracterÃsticas fenotÃpicas de C. posadasii e desenvolver mÃtodos independentes de cultivo para o diagnÃstico laboratorial da coccidioidomicose. O estudo fenotÃpico de 10 cepas (7 isolados clÃnicos e 3 isolados ambientais) foi conduzido por anÃlises macro e micromorfolÃgica, anÃlise da cinÃtica de crescimento frente a estresse salino, tÃrmico e de pH; testes de assimilaÃÃo de fontes de carbono e nitrogÃnio e determinaÃÃo do perfil de sensibilidade frente a antimicrobianos. Adicionalmente, foram avaliados mÃtodos de extraÃÃo de antÃgenos de cepas locais de C. posadasii e amplificaÃÃo de seqÃÃncias genÃticas exclusivas do fungo em escarro. Foram observadas trÃs variaÃÃes morfolÃgicas das cepas de C. posadasii cultivadas em Ãgar batata, Ãgar Sabouraud dextrose a 2% e Ãgar YEG. Com relaÃÃo a micromorfologia, foi detectado que a quantificaÃÃo de artroconÃdios estava relacionada à idade e a morfologia das colÃnias. A avaliaÃÃo da cinÃtica de crescimento das cepas, in vitro, revelou que as cepas de C. posadasii sÃo inibidas por elevada concentraÃÃo salina e temperaturas acima de 40 &#61616;C, mas nÃo sofrem influÃncia quanto à variaÃÃo de pH do meio. A anÃlise nutricional demonstrou que vÃrios compostos, inclusive os Ãons oxidados nitrito e nitrato, podem ser utilizados como fontes de carbono e/ou nitrogÃnio. Todas as cepas de C. posadasii foram sensÃveis Ãs drogas antifÃngicas anfotericina B, caspofungina e aos derivados azÃlicos cetononazol, itraconazol, fluconazol e voriconazol. Foi observado que, a exceÃÃo da pirazinamida, todos as demais drogas antituberculose alteraram o crescimento das cepas de C. posadasii in vitro. CombinaÃÃes formadas pelos quimioterÃpicos antituberculose tambÃm mostraram efeito inibitÃrio sobre o crescimento das cepas. Os protocolos experimentais culminaram com uma preparaÃÃo antigÃnica estÃvel e imunoreativa, adequada para utilizaÃÃo em testes sorolÃgicos presuntivos para diagnÃstico da coccidioidomicose. A identificaÃÃo molecular de C. posadasii foi realizada, com sucesso, a partir da amplificaÃÃo da seqÃÃncia do gene pra diretamente de culturas filamentosas e escarro contendo estruturas parasitÃrias do fungo. Acredita-se que as caracterÃsticas fenotÃpicas descritas para as cepas de C. posadasii poderÃo auxiliar estudos futuros que objetivem a individualizaÃÃo biolÃgica das espÃcies C. immitis e C. posadasii. Os resultados desse estudo contribuÃram para o estabelecimento de estratÃgias para o maior conhecimento da coccidioidomicose no BrasilCoccidiodomycosis is a systemic infection caused by the dimorphic fungi species Coccidioides immitis and C. posadasii. The disease affects both humans and animals, and is considered endemic in the semiarid areas of Cearà State (Northeast Brazil), where it is caused by C. posadasii. The aims of this study were to analyse the phenotypical characteristics of C. posadasii and to develop culture independent methods for improving of the laboratorial diagnosis of coccidioidomycosis. The phenotypical study was conducted with 10 strains (7 from clinical and 3 from ambiental sources) by macro and micromorphology analyses; in vitro growth under different conditions of salinity, temperature and pH; nutritional analysis and antimicrobial susceptibility tests. In addition, methods of antigens extraction and amplification of C. posadasii DNA directly in sputum were also tested. The mycological analysis showed three morphological variations of C. posadasii strains cultivated in potato agar, Sabouraud agar and YEG agar. Regarding to micromorphology, it was observed that arthroconidia counts were related to colony aging and texture. Growth rates of C. posadasii strains were inhibited by high salt concentrations, temperatures above 40 C, but were not affected by the range of pH. Nutritional analyses showed that several compounds, including the oxidized ions, nitrite and nitrate, were efficiently metabolized by C. posadasii strains as carbon and/ or nitrogen sources. The antifungal susceptibility analysis showed that all of the strains of C. posadasii were sensitive to amphotericin B, caspofungin and the azoles ketoconazole, itraconazole, fluconazole, and voriconazole. With the exception of pyrazinamide, all of the tested drugs interfered with the in vitro growth of C. posadasii. Combined antituberculosis drugs also inhibit fungal growth in vitro. Experimental protocols produced a stable and immunoreactive antigen, suitable in presumptive serologic tests for the diagnosis of coccidioidomycosis. Molecular identification of C. posadasii was achieved by PCR amplification of the specific pra gene directly from filamentous cultures and sputum with fungal parasitic structures. The data obtained by phenotypical analyses may be useful in future studies for biological distinction of C. immitis and C. posadasii species. The results of this study provided approaches for a better understanding of coccidioidomycosis in Brazi

Successive mycological nail tests for onychomycosis: a strategy to improve diagnosis efficiency

Onychomycosis is a fungal infection of nails caused by dermatophytes, yeasts and moulds, accounting for about 50% of onychopathies. A high frequency of onychomycosis caused by Candida species has been reported during the last few years in northeast Brazil, as well as in other regions of the world. A clinical diagnosis of onychomycosis needs to be confirmed through laboratory exams. We evaluated the importance of serial repetition of direct microscopic exams and fungal culture for the diagnosis of onychomycosis in the city of Fortaleza, Ceará, in northeast Brazil. We first made a retrospective study of 127 patients with onychomycosis, identifying the fungi that had been isolated from fingernails and toenails. We then made a prospective study of 120 patients, who were submitted to three successive mycological examinations. Ungual residues were scraped off and directly examined with a microscope and fungal cultures were made. In the retrospective study, in which only one sample was analyzed, the incidence of onychomycosis was 25.0%. In our prospective study, in which we had data from successive mycological examinations, 37.8% had onychomycosis. The most commonly isolated fungi in both studies were yeasts from the genera Candida, especially C albicans, C. parapsilosis and C. tropicalis. We found a high proportion of onychomycosis caused by Candida species. We also concluded that serial repetition of direct microscopic examination and fungal culture, with intervals of 2-5 days improved the diagnosis of onychomycosis. We suggest that this laboratorial strategy is necessary for accurate diagnosis of this type of mycosis, especially when the standard procedures fail to diagnose fungal infection, despite strong clinical suspicion

Candidose na medicina veterinária: um enfoque micológico, clínico e terapêutico Candidosis on veterinary medicine: a mycological, clinical and therapeutic approuch

O gênero Candida é composto por leveduras que vivem como comensais na microbiota de homens e animais. Em geral, não causam nenhum dano aos seus hospedeiros, entretanto, em virtude de desequilíbrios nas defesas química, física e imunológica, esses microrganismos podem se tornar patogênicos. Infecções por Candida spp. são pouco frequentes na Medicina Veterinária no entanto, nos últimos anos, tem sido observado aumento considerável de relatos de enfermidades causadas por essas leveduras, acometendo diferentes animais. Várias espécies do gênero são implicadas em quadros infecciosos, sendo a C. albicans a principal delas, seguida por C. tropicalis e C. parapsilosis. Considerando-se o potencial patogênico do gênero Candida, aliado ao surgimento de cepas resistentes a derivados azólicos, in vitro, o presente trabalho se propôs a realizar detalhada revisão de literatura, abordando os aspectos clínico-laboratoriais, etiológicos e terapêuticos da candidose na Medicina Veterinária.<br>The Candida genus is composed by yeasts that live as commmensal on human and animals' microbiota. In general, they do not cause any damage to their hosts. However, due instability on chemical, physical and immunological defenses, these microorganisms can become pathogens. Candida spp. infections are rare on Veterinary Medicine. However, on the last years, a considerable raise of illness caused by these yeasts has been related on varied animal species. Several species of this genus has been mentioned as responsible for infectious diseases in animals, being C. albicans the main of them, followed by C. tropicalis and C. parapsilosis. Considering the pathogenic role of the genus Candida, allied to the emerging of resistant strains to the azole derivatives, in vitro, the present research proposed to perform a detailed review, approaching clinic-laboratorial, etiologic and therapeutic aspects of the candidosis on Veterinary medicine

CANDIDEMIA IN A BRAZILIAN HOSPITAL: THE IMPORTANCE OF Candida parapsilosis

SUMMARY The aim of this study was to perform a retrospective analysis of cases of candidemia in a Brazilian hospital in the city of Fortaleza, Ceará. A total of 50 blood cultures were analyzed from 40 candidemic patients. The mycological diagnosis was based on the phenotypical analysis and the patients&apos; data were recorded in appropriate files. The most frequent species were Candida parapsilosis (n = 18), followed by C. albicans (n = 14), C. tropicalis (n = 8), C. guillermondii (n = 6), C. glabrata (n = 2), and Candida spp. (n = 2). A detailed descriptive study was undertaken with 21 patients whose medical records were complete. The candidemia episodes occurred in eight male patients and 13 female patients. The most representative risk factors implicated in candidemia were prior antibiotic therapy, central venous catheters, parenteral nutrition, gastric probes and mechanical ventilation. Death occurred in 13 of the 21-candidemic patients. This study demonstrated the emergence of candidemia caused by C. parapsilosis in a Brazilian hospital in the city of Fortaleza, Ceará