Imidazolium Salts for Candida spp. Antibiofilm High-Density Polyethylene-Based Biomaterials.

The species of Candida present good capability to form fungal biofilms on polymeric surfaces and are related to several human diseases since many of the employed medical devices are designed using polymers, especially high-density polyethylene (HDPE). Herein, HDPE films containing 0; 0.125; 0.250 or 0.500 wt% of 1-hexadecyl-3-methylimidazolium chloride (C16MImCl) or its analog 1-hexadecyl-3-methylimidazolium methanesulfonate (C16MImMeS) were obtained by melt blending and posteriorly mechanically pressurized into films. This approach resulted in more flexible and less brittle films, which impeded the Candida albicans, C. parapsilosis, and C. tropicalis biofilm formation on their surfaces. The employed imidazolium salt (IS) concentrations did not present any significant cytotoxic effect, and the good cell adhesion/proliferation of human mesenchymal stem cells on the HDPE-IS films indicated good biocompatibility. These outcomes combined with the absence of microscopic lesions in pig skin after contact with HDPE-IS films demonstrated their potential as biomaterials for the development of effective medical device tools that reduce the risk of fungal infections.

A chloroacetamide derivative as a potent candidate for fusariosis treatment.

Fusariosis has presented a significant increase in their incidence in the last years. This epidemiological panorama probably is due to the increasing profile of refractory susceptibility of Fusarium spp. to available drugs, especially in immunocompromised individuals. Thus, the development of new compounds with effectiveness on these organisms is a necessity. This study evaluated the antifungal potential of a chloroacetamide derivative (4-BFCA) against resistant Fusarium strains. As a result, the compound was effective against all strains (MIC range 12.5-50 µg/mL). The time kill assay demonstrated that 4-BFCA presents a concentration-dependent fungicidal action. Although its action mechanism has not yet been elucidated, it was possible to observe its efficacy through damages and alterations provoked along the hyphae of Fusarium spp. 4-BFCA maintained a high survival rate of Tenebrio molitor larvae, suggesting that it does not cause acute systemic toxicity on this host at the concentration evaluated. In addition, 4-BFCA was 83.33% effective in combating a fungal infection in vivo on the chorioallantoid membrane of embryonated eggs. Our results are very promising and arouse interest to investigate the action of 4-BFCA on Fusarium strains since it acts as a possible candidate for the development of new therapies for the treatment of fusariosis.

Discovery of a novel and selective fungicide that targets fungal cell wall to treat dermatomycoses: 1,3-bis(3,4-dichlorophenoxy)propan-2-aminium chloride.

BACKGROUND: Fungal infections are highly prevalent and are responsible for high rates of morbidity and mortality. In this context, the search for new treatment alternatives is very relevant. OBJECTIVES: Analyse chemical compounds for antifungal potential against dermatomycosis fungi. METHODS: The antifungal activity of 121 compounds, intermediates or derivatives of 1,3-bis(aryloxy)propane substituted at C-2 (111 compounds) and isothiouronium derivatives (10 compounds) was investigated through susceptibility tests, mechanism of action, toxicity and hydrogel incorporation. RESULTS: The compound 1,3-bis(3,4-dichlorophenoxy)propan-2-aminium chloride (2j) was the most active fungicide against dermatophytes and Candida spp., at very low concentrations (0.39-3.12 µg/mL), including action on resistant and multidrug-resistant clinical strains. Compound 2j has presented a promising toxicity profile, showing selectivity index >10, relative to human lymphocytes. The compound was classified as non-irritant by the HET-CAM test and did not cause histopathological alterations in pig ear skin, thus presenting an excellent perspective for topical application. 2j targets the fungal cell wall, which was confirmed by scanning electron microscopy, which also indicated the additional ability of 2j to inhibit the Candida albicans pseudohyphae formation and biofilm of Microsporum canis. Compound 2j was incorporated in a hydrogel with bioadhesive potential. The results of the human skin permeation showed that 2j remained significantly in the epidermis, ideally for the dermatomycosis treatment. CONCLUSIONS: Therefore, the compound 2j demonstrated the potential for antifungal drug development, with a action mechanism elucidated and already applied in a semisolid formulation as a new therapeutic option for fungal skin infections.

Epidemiological aspects and characterization of the resistance profile of Fusarium spp. in patients with invasive fusariosis.

Introduction. The remarkable intrinsic resistance of Fusarium species to most antifungal agents results in high mortality rates in the immunocompromised population.Aims. This study aimed to investigate the epidemiology, clinical features and antifungal susceptibility of Fusarium isolates in patients with invasive fusariosis.Methodology. A total of 27 patients admitted to a referral hospital from January 2008 to June 2017 were evaluated. Antifungal susceptibility testing of isolates was performed by broth microdilution according to the Clinical and Laboratory Standards Institute guidelines.Results. Haematological malignancy was the predominant underlying condition, with an incidence of invasive fusariosis of 14.8 cases per 1000 patients with acute lymphoid leukaemia and 13.1 cases per 1000 patients with acute myeloid leukaemia. The Fusarium solani species complex (FSSC) was the most frequent agent group, followed by the Fusarium oxysporum species complex (FOSC). Voriconazole showed the best activity against Fusarium, followed by amphotericin B. Itraconazole showed high minimum inhibitory concentration values, indicating in vitro resistance. Clinical FSSC isolates were significantly (P<0.05) more resistant to amphotericin B and voriconazole than FOSC isolates.Conclusion. The present antifungal susceptibility profiles indicate a high incidence of fusariosis in patients with haematological malignancy. Species- and strain-specific differences in antifungal susceptibility exist within Fusarium in this setting.

Fungal infection models: Current progress of ex vivo methods.

Experimental alternative ex vivo models that simulate infectious processes in vivo are of fundamental importance for the evaluation of new drugs, since in some cases, their execution does not depend on the approval of an ethics committee in research. Although studies using alternative infectious models to evaluate the efficacy of antifungal molecules have been increasingly described and reported, there is no critical consensus that establishes the most appropriate ones regarding the type of infection. Numerous studies contemplate ex vivo protocols of fungal infections on nails, corneas, dentinal tubules and skin and reveal counterpoints and concordances not yet finely confronted. In this minireview, we propose a critical analysis of the main ex vivo models of fungal infections for the evaluation of new antifungal candidates for both topical and systemic use, as opposed to the advantages and disadvantages of the traditional in vivo models employed in preclinical research.

Lipid core nanoparticles as a broad strategy to reverse fluconazole resistance in multiple Candida species.

Fungal resistance is the major problem related to fluconazole treatments. This study aims to develop innovative lipid core nanocapsules and nanostructured lipid carriers containing fluconazole, to study in vitro antifungal activity and to assess the possibility of resistance reversion in Candida albicans, C. glabrata, C. krusei, and C. tropicalis isolates. The action mechanism of nanoparticles was investigated through efflux pumps and scanning electron microscopy studies. The lipid core nanocapsules and nanostructured lipid carriers were prepared by interfacial deposition of preformed polymer and high-pressure homogenization methods, respectively. Both nanostructures presented sizes below 250 nm, SPAN < 1.6, negative zeta potential, pH slightly acid, high drug content and controlled drug release. The nanostructured lipid carriers were unable to reverse the fungal resistance. Lipid core nanoparticles displayed advantages such as a reduction in the effective dose of fluconazole and resistance reversion in all isolates tested - with multiple mechanisms of resistance. The main role of the supramolecular structure and the composition of the nanoparticles on antifungal mechanisms of action were discussed. The results achieved through this study have an impact on clinical therapy, with a potential application in the treatment of fungal infections caused by resistant isolates of Candida spp.

Perfil de inaptidão na triagem clínica e sorológica de candidatos à doação de sangue / Profile of inability in the clinic and serological screening of candidates for blood donation

Objetivo: Caracterizar os candidatos quanto ao gênero e analisar a prevalência dos critérios de inaptidão adotados às doações de sangue, realizadas no Banco de Sangue Santa Maria, em Santa Maria, RS. Métodos: Dados retrospectivos arquivados foram avaliados, analisando-se os critérios de exclusão na triagem clínica e sorológica, no período de janeiro de 2005 até dezembro de 2010. Foram analisados 20.264 doadores. O estudo foi aprovado pelo Comitê de Ética e Pesquisa da Universidade Federal do Rio Grande do Sul-UFRGS, sob número de protocolo 18147. Resultados: Novecentos e setenta e seis (5%) candidatos foram considerados inaptos pela triagem clínica e 19.288 (95%) foram considerados aptos para prosseguirem com os testes laboratoriais. Dos doadores aptos, 941 (5%) foram excluídos na triagem sorológica, totalizando 18.347 bolsas de sangue disponíveis para uso. Predominaram os doadores do sexo masculino (62%) e a principal causa de exclusão, na triagem clínica, foi hipertensão (0,7%). Para a triagem sorológica, a principal causa de exclusão foi a Doença de Chagas (1,5%). Conclusão: Ressaltou-se a importância da triagem clínica, tendo em vista que ela excluiu 5% dos candidatos à doação e a relevância da triagem sorológica ser feita corretamente, evitando que resultados falso-negativos sejam liberados.

Objective: Characterize donors by gender and analyze theprevalence of the inability criteria of donations made at the SantaMaria Blood Bank, located at Santa Maria, Brazil. Methods: First,we searched historical data for exclusion criteria used in clinicaland serological screenings during the period from January, 2005 toJuly, 2010. We evaluated 20,264 blood donors' data for this study.The Ethical Committee of Rio Grande do Sul Federal UniversityUFRGSapproved this study under the protocol number 18147.Results: Nine hundred and seventy-six (5%) candidates wereconsidered unfit by the clinical screening and 19,288 (95%) wereconsidered fit. From the resulting fit population of the clinicalscreening, 941 (5%) were excluded from donating by the serologicalscreening, totaling 18.347 blood donations fit for use. The resultsshows that majority of the donors were male (62%) and the leadingcause of exclusion from donating in the clinical screening washypertension (0.7%). As for the serological screening, the leadingcause for exclusion was Chagas disease (1.5%). Conclusion: Thestudy stress the importance of the clinical screening process, giventhat it excluded 5% of blood donations, which were unfit for use. Wealso notice the relevance of a correctly done serological screening,thus avoiding that false-negative results are released.

Etiología de la candidiasis vulvovaginal recidivante en la Atención Primaria de Salud en Santa Catarina, Brasil / Etiology of recurrent vulvovaginal candidiasis in the National Health System in Santa Catarina, Brazil / Etiologia da candidíase vulvovaginal recorrente na Atenção Primária à Saúde em Santa Catarina, Brasil

El objetivo del presente trabajo fue destacar las características epidemiológicas que puedan subsidiar la Atención Primaria de Salud (APS) en mujeres portadoras de candidiasis vulvovaginal (CVV) y candidiasis vulvovaginal recidivante (CVVR), a partir de estudios realizados en tres municipios del sur de Brasil. A través del examen micológico de la secreción vaginal de 300 mujeres con sospecha clínica de CVV o CVVR se identificaron las especies prevalentes de Candida, correlacionándose los hallazgos con los principales factores de riesgo mencionados en la literatura. Fueron confirmadas levaduras en 90 (30%) casos, resultando las especies más frecuentes C. albicans (61,1%), C. krusei (16,7%), C. tropicalis (6,7%), C. glabrata (4,4%) y Candida spp. (11,1%). En los casos de CVVR, C. albicans fue la especie más encontrada, con una prevalencia superior a la observada en la CVV. C. krusei apareció como la segunda especie más prevalente en todas las muestras, resaltando la importancia del diagnóstico a nivel de especie, dada la resistencia intrínseca al fluconazol. Las informaciones epidemiológicas del estudio son útiles para que los gestores de la Atención Primaria de Salud (APS) y los profesionales de la Salud puedan tener subsidios adicionales para actuar preventivamente en el caso de candidiasis vulvovaginales.

The main purpose of this work was to highlight epidemiological characteristics serving as subsidies to health promotion activities for vulvovaginal candidiasis (VVC) and recurrent vulvovaginal candidiasis (RVVC) by the national health system, in three cities in southern Brazil. Through the mycological examination of vaginal secretions of 300 women with clinical suspicion of VVC or RVVC, Candida-prevalent species were identified and they were correlated with the main risk factors mentioned in the literature. Yeasts were confirmed in 90 (30%) cases, resulting in C. albicans 61.1%, C. krusei 16.7%, C. tropicalis 6.7%, C. glabrata 4.4% and others 11.1%. C. albicans was the species most commonly found in cases of RVVC, with levels higher than the prevalence of the species in the VVC. C. krusei prevailed as the second most prevalent species in both samples, emphasizing the importance of diagnosing the species level, due to its intrinsic resistance to fluconazole. The epidemiological information of the study is useful for managers of the National Health Care System, as well as direct health professionals, who can have new subsidies to act preventively against vulvovaginal candidiasis.

O objetivo do presente trabalho foi destacar as características epidemiológicas que possam subsidiar a Atenção Primária à Saúde (APS) em mulheres portadoras de candidíase vulvovaginal (CVV) e candidíase vulvovaginal recorrente (CVVR), a partir de estudos realizados em três municípios do sul do Brasil. Através do exame micológico da secreção vaginal de 300 mulheres com suspeita clínica de CVV ou CVVR foram identificadas as espécies prevalentes de Candida, correlacionando os achados com os principais fatores de risco mencionados na literatura. Foram detectadas leveduras em 90 (30%) dos casos, resultando as espécies mais frequentes C. albicans (61,1%), C. krusei (16,7%), C. tropicalis (6,7%), C. glabrata (4,4%) e Candida spp. (11,1%). Nos casos de CVVR, C. albicans foi a espécie mais encontrada, com uma prevalência superior à observada nos casos de CVV. C. krusei apareceu como a segunda espécie mais prevalente em todas as amostras, ressaltando a importância do diagnóstico em nível de espécie, devido à resistência intrínseca ao fluconazol. As informações epidemiológicas deste estudo são úteis para que os gestores da Atenção Primária à Saúde (APS) e os profissionais da Saúde Pública possam ter subsídios adicionais para atuar preventivamente nos casos de candidíases vulvovaginais.

Occurrence and identification of yeasts in dogs external ear canal with and without otitis / Ocurrencia y detección de levaduras en el conducto auditivo de los perros con y sin otitis

Objective. To analyze the presence of yeast in the external ear canal of 116 dogs with and without a diagnosis of otitis from veterinary clinic in the Chapecó city, Santa Catarina, Brazil, and to examine the secretion of the proteinase in isolates. Materials and methods. Were collected cerumen of conduct hearing of dogs of 16 different races 71% with pendular ear type, 5% of semi-pendular and 24% of the erect type. All dogs were previously evaluated by otoscopy and grouped in dogs with and without otitis. Results. Yeasts were isolated in 44 samples (approximately 36%), where Malassezia pachydermatis was identified in 95% of samples where were observed growth of yeasts. On 20 samples the proteinase enzyme showed strong activity in 31% isolates, were 21% of the dogs with otitis tested showed high proteolytic activity. Conclusions. We observed a variation of strains of M. pachydermatis-producing enzymes. The variation in production of these enzymes is probably more associated with different response to the action of the immune system of the animal in the tissue injury.

Objetivo. Se investigó la presencia de levaduras en el canal externo del oído de 116 perros de la clínica veterinaria en la ciudad de Chapecó, Santa Catarina,Brasil, en perros sanos y perros con otitis y se examinó la secreción de la proteinasa en las muestra aisladas. Materiales y métodos. Se recogieron cerumen del oído de perros de 16 razas diferentes, dónde 71% fue de oído de tipo pendular, 5% de semi-pendular y 24% del tipo erecto. Todos los perros fueron evaluados previamente por otoscopia y agrupados en perros con y sin otitis externa. Resultados. Las levaduras se aislaron en 44 muestras (aproximadamente 36%), donde Malassezia pachydermatis se identificó en el 95% de las muestras donde se observó el crecimiento de las levaduras. El 20 muestras la secreción de proteinasa mostró fuerte actividad en el 31% de los aislados y en 21% de los perros con otitis mostró alta actividad proteolítica. Conclusiones. Hemos observado una variación de cepas de M. pachydermatis productoras de enzimas. La variación en la producción de estas enzimas es probablemente más asociados con la respuesta diferente a la acción del sistema inmunológico del animal en la lesión tisular.

Chromoblastomycosis in Santa Catarina state, Brazil / Cromoblastomicosis en el estado de Santa Catarina, Brasil

A case of chromoblastomycosis caused by Cladophialophora carrionii is reported. The diabetic and hypertensive patient presented serpiginous and verrucous lesions, with centrifugal evolution. The patient, with a history of disease for 59 years, had not been diagnosed or treated before. Dematiaceous septate hyphal and elliptical conidia were seen on microscopic observations. The isolated fungus was identified on the basis of micro-macromorphologic characteristics.

Se reportó un caso de cromoblastomicosis causado por Cladophialophora carrionii. El paciente, diabético e hipertenso, presentaba lesiones de apariencia verrugosa y serpiginosa, con evolución centrífuga. Tenía un historial de enfermedad hace 59 años sin haber sido diagnosticado hasta entonces. En el examen microscópico de observaron elementos hifales septados, pigmentados y con conidios elípticos. La identificación del hongo se basó en las características macromorfológicas y micromorfológicas.

Determinación de zearalenona por cromatografía líquida de alta resolución: normalización de un nuevo método para raciones especiales para cerdos / Zearalenone determination in high performance liquid chromatography: a new standardization method for commercial swine rations

The aim of this work was to offer a new method of high performance liquid chronomatography (HPLC) to evaluate commercial swine rations (CSR) contaminated by zearalenone (ZEA). After ZEA extraction and purification from CSR, the samples were eluted with acetonitrile, methanol and water solvent system. The results indicated that the proposed method showed to be rapid and efficient for the detection and quantification of ZEA in CSR, since its recovery was 102.62 percent, it offered excellent precision with a coefficient of variation of 0.9992. Furthermore, it is also proposed a as a biocontrol assay for micotoxigenic fungi isolated and maintained in the laboratory. The test was performed with the killer yeast Trichosporum insectorum CBS 10422 against Fusarium sp and Aspergillus flavus, which demonstrated to be effective against the latter.

El propósito de este articulo es ofrecer un nuevo método de cromatografía líquida de alta resolución (CLAR) para evaluar las raciones especiales para cerdos (REC) contaminado con zearalenona (ZEA). Después de la extracción y purificación de ZEA, las muestras se eluyeron con acetonitrilo, metanol y agua del sistema disolvente. Los resultados indican que el método propuesto demostró ser rápido y eficaz para la detección y cuantificación de ZEA en REC, ya que sus indicadores se presentan capaces de recuperación de 102,62 por ciento, además de ofrece una excelente precision, con un çõefiCiênte de variación de 0,9992. Por otra parte, también se propone una prueba de control biológico de hongos micotoxige-nic aislados y mantenidos en el laboratorio. La prueba se realizó con la levadura killer Trichosporum insectorum CBS 10422 contra Fusarium sp y Aspergillus flavus, mostrando eficaces sólo contra Aspergillus.

Chromoblastomycosis in Santa Catarina state, Brazil.

A case of chromoblastomycosis caused by Cladophialophora carrionii is reported. The diabetic and hypertensive patient presented serpiginous and verrucous lesions, with centrifugal evolution. The patient, with a history of disease for 59 years, had not been diagnosed or treated before. Dematiaceous septate hyphal and elliptical conidia were seen on microscopic observations. The isolated fungus was identified on the basis of micro-macromorphologic characteristics.

Validity of method for determining deoxynivalenol through high performance liquid chromatography with detection in the ultraviolet in wheat samples / Validación de un método para la determinación de deoxinivalenol por cromatografía líquida de alta resolución con deteccíon ultravioleta en muestras de trigo

With the purpose of evaluating the wheat grain and wheat flour contamination by Deoxynivalenol (DON) in the municipality of Chapeco-SC, and standardize an useful method to detect this mycotoxin by High Performance Liquid Chromatography (HPLC), six samples of wheat grains from different storage location, and one sample of wheat grain from the flour milling industry were obtained during in the month of august 2008. Samples belong to a corporation from Chapeco-SC that stores and processes wheat grains, flour and wheat middlings, among other products. The extraction has been carried out with methanol: water (100: 100 v / v), filtered paper filter and applied in immunoaffinity column specific DON. After the wash with water column, the toxin was eluted with methanol. The detection and quantification Deoxynivalenol in samples was carrid out through the method of HPLC in the UV-visible with detection 244 nm. The 6 analyzed samples of wheat grain showed DON levels within 7.0 and 10.1 ppb, while the wheat flour contained 90.2 ppb. DON contents in wheat grains and wheat flour are lower than the limits claimed by the studied corporated importers and the international legislation.

Con el objetivo de evaluar la contaminación por Deoxinivalenol (DON) en granos y harina del trigo en la municipalidad de Chapeco-SC y estandarizar un método de deteccíon para este micotoxina por cromatografía líquida de alta resolución (CLAR), se procesaron durante el mes de agosto de 2008, seis muestras de granos del trigo en diferentes situaciones de almacenamiento y una muestra de harina de trigo de un molino . Las muestras pertenecen a una cooperativa de Chapeco-SC que procesa y almacena granos y harinas de trigo entre otros productos. La extracción de la micotoxina se obtuvo con metanol: agua (100: 100 v / v), filtrado en papel filtro y aplicado a una columna de inmunoafinidad específica (DON). Después del lavado de la columna con agua, la toxina fue elucidada con metanol. La detección y cuantificación de Deoxinivalenol en las muestras se determinó por el método CLAR en el UV- visible con una longitud de onda de 244 nm. Las 6 muestras analizadas del grano, mostraron que el DON nivela entre 7,0 y 10, 1 ppb, mientras la harina del trigo alcanzó las 90,2 ppb. Los niveles de DON en los granos y harina de trigo tienen límites menores que los exigidos por las cooperativas importadoras estudiadas y la legislación internacional.