Histomorphometric comparison of left and right oviduct structure from alpaca (Vicugna pacos).

Alpacas are species of induced ovulation and with foetal development only in the left uterine horn (98%). The histoarchitecture of the oviductal regions determine a spatio-temporal interaction between the gametes/embryos and the oviduct. This study compares the morphometric changes of the left and right oviducts in alpaca during the follicular phase. Five oviducts (n = 05), from adult alpacas with dominant follicle in the right ovary were recovered, dissected, and processed by histological technique with H&E and PAS stain for measurement of morphometric parameters and cell characteristics, respectively. Also, a 3D image reconstruction was performed (by reconstruct software). Resin moulds (polyurethane PU4ii) were applied for visualization of oviductal lumen. The multivariable data of parameters were analysed with ANOVA and principal component analysis (PCA). The histomorphometric parameters of left and right oviducts did not show statistically significant differences (p ≥ 0.05), although PCA showed morphometric differences between regions of the oviduct. No differences were observed between the 3D reconstruction of the left and right oviducts, as well as in the luminal spaces examined in the resin moulds. In conclusion, the histomorphometry of the oviduct is not affected by its location on either the left or right side; therefore, it cannot explain why 98% of foetuses implant in the left uterus.

Modeling the inactivation of Aspergillus fischeri and Paecilomyces niveus ascospores in apple juice by different ultraviolet light irradiances.

The present work aimed to evaluate and to model the influence of UV-C light treatments with different irradiances (6.5, 13, 21, and 36 W/m2) on Aspergillus fischeri and Paecilomyces niveus ascospores inactivation in clarified apple juice. Approximately 5.0 and 6.0 log CFU/mL spores of P. niveus and A. fischeri, respectively, were suspended in 30 mL of clarified apple juice (pH 3.8, 12 ± 0.1°Brix) and exposed to UV-C light at different irradiances (as above) and exposure times (0 to 30 min). The first-order biphasic model was able to describe the experimental data with good statistical indices (RMSE = 0.296 and 0.308, R2 = 0.96 and 0.98, for P. niveus and A. fischeri respectively). At the highest irradiance level tested (36 W/m2), the UV-C light allowed the reduction of 5.7 and 4.2 log-cycles of A. fischeri and P. niveus ascospores, respectively, in approximately 10 min. P. niveus was the most UV-C resistant mould. The results showed that, to a defined UV-C fluence, a change in the level of either time or UV-C irradiance did not affect the effectiveness of UV-C light for A. fischeri and P. niveus inactivation. Thus, the modeling of the inactivation as a function of the UV-C fluence allowed the estimation of the primary model parameters with all experimental data and, consequently, no secondary models were needed. The model parameters were validated with experiments of variable UV-C fluences. Accordingly, experimental results allowed to conclude that UV-C treatment at the irradiances tested is a promising application for preventing A. fischeri and P. niveus spoilage of juices.

Silage fermentation-updates focusing on the performance of micro-organisms.

Advances in micro-organism identification techniques have resulted in increased knowledge of the diversity of prokaryotes and eukaryotes in silage. Such knowledge has enhanced the understanding of how fermentation occurs in forage crops with different characteristics and how the process can be improved to enhance silage quality. Undesirable micro-organisms can grow in silage when fermentation does not occur properly. Such micro-organisms may be pathogenic and/or produce toxic metabolic compounds; however, information on the consequences of these metabolites on the health of animals that consume silage is still lacking. The major challenge of ensilage is to produce high-quality feed that is nutritional, sanitary and stable, with a high dry matter recovery rate, in a process involving no interventions during fermentation and considerable variation in the characteristics of the substrates. It is important to note that each substrate has particularities and that we can only improve fermentation if we fully understand microbial diversity. This review is intended to update information related to the fermentation profile of silage, focusing on microbial diversity.

Epidemiología de la enfermedad fúngica invasora por hongos filamentosos en el período 2005 a 2015, en un hospital universitario en Santiago, Chile / Epidemiology of invasive fungal disease by filamentous fungi in the period 2005 to 2015, in a university hospital in Santiago, Chile

Resumen Introducción: La enfermedad fúngica invasora (EFI) por hongos filamentosos es cada vez más frecuente. Objetivo: Estudiar la epidemiología de la EFI en adultos hospitalizados en nuestro centro. Metodología: Estudio retrospectivo de pacientes adultos de un hospital universitario en Santiago, Chile, con EFI por hongos filamentosos entre enero de 2005 y diciembre de 2015. Resultados: Se identificaron 125 episodios, siendo 48% categoria probada, 40% probable y 11% posible según criterios EORTC/MSG, incidencia global 0,47 x 1.000 egresos, 57% pacientes masculinos y edad de 50 ± 16 años. El 66,4% tenía patología hematológica, 11,2% trasplante de órgano sólido, 11,2% enfermedad reumatológica, 11,2% otra condición. Los factores de riesgo fueron neutropenia 44%, corticoterapia 21%, inmunosupresores 13%. Los hongos más frecuentemente identificados fueron Aspergillus spp (53,6%), Mucorales (16%), Fusarium spp (8,8%), Alternaria spp (5,6%), otros filamentosos (3,2%). Todos recibieron antifúngicos, 82% monoterapia, 18% terapia combinada, hubo defocación quirúrgica en 90% de mucormicosis. La mortalidad global fue 42%. Al comparar 2005-2009 vs 2010-2015, hubo un aumento significativo de la incidencia y una tendencia a menor mortalidad en el segundo período. Conclusiones: Durante un período de 10 años, observamos un aumento de la incidencia de EFI por filamentosos, aspergilosis fue la etiología más frecuente y la mortalidad global fue 42%.

Background: Invasive fungal disease (IFD) due to filamentous fungi is increasingly common. Aim: To study the epidemiology of EFI in hospitalized adults in our center. Methods: Retrospective study of adult patients of a university hospital in Santiago, Chile, with EFI due to filamentous fungi between January 2005 and December 2015. Results: 125 episodes were identified, being 48% proven, 40% probable and 11% possible according to EORTC/MSG criteria, overall incidence was 0.47/1,000 admissions, 57% male patients and age 50 ± 16 years. 66.4% had hematological pathology, 11.2% solid organ transplant, 11.2% rheumatology diseases, 11.2% other conditions. The risk factors were neutropenia 44%, corticosteroid therapy 21%, immunosuppressants 13%. The most frequent mould identified were Aspergillus spp (53.6%), Mucorales (16%), Fusarium spp (8.8%), Alternaria spp (5.6%) and other filamentous (3.2%). All received antifungals, 82% monotherapy, 18% combined therapy, there was surgical defocation in 90% of mucormycosis. The overall mortality was 42%. When comparing 2005-2009 vs 2010-2015, there was a significant increase in incidence and a tendency to lower mortality in the second period. Conclusions: Over a period of 10 years, we observed an increase in the incidence of EFI by filamentous, aspergillosis was the most frequent etiology and the overall mortality was 42%.

Social, environmental and microbiologic aspects of endemic mycoses in Brazil.

Many factors that lead to host immunosuppression are clearly known to predispose the host to fungal diseases, significantly influencing the occurrence of mycoses. However, little or nothing has been discussed regarding social or economic factors that can influence the occurrence of diseases caused by fungi. In this minireview, we discuss several factors that may affect the occurrence of mycoses in Brazil, a continentally extended country that is marked by large climatic variations and severe socioeconomic distortions that may limit access to health services for the population.

Micobiota do Queijo Artesanal Serrano produzido em Santa Catarina, Brasil / Mycobiota of Serrano Artisanal Cheese produced in Santa Catarina, Brazil

O Queijo Artesanal Serrano é um produto nativo do sul do Brasil, produzido por mão de obra familiar a partir do leite cru, e que vem buscando a obtenção da sua indicação geográfica. Muitas das características que particularizam os tipos de queijo são dadas pela sua micobiota natural. O objetivo deste estudo foi determinar as espécies fúngicas presentes em 20 amostras de Queijo Artesanal Serrano de quatro períodos de maturação (14, 21, 28 e 35 dias), produzidos em outubro de 2017, em Santa Catarina. A identificação dos bolores foi feita por chaves de identificação e das leveduras por MALDI-TOF. Os bolores foram isolados em 28 das 80 amostras e com espécies variáveis, sendo algumas contaminantes. Por outro lado, as leveduras foram detectadas em todas as amostras, com alguns gêneros mais frequentes, principalmente o Kluyveromyces lactis, presente em 27 amostras, e algumas espécies de Candida spp. As espécies identificadas já foram isoladas em outros queijos artesanais brasileiros, produzido em locais com características de clima e relevo semelhantes ao do Queijo Artesanal Serrano.(AU)

Serrano Artisanal Cheese is a native product from the South region of Brazil, produced by family labor from raw milk, which seeks to obtain its geographical indication. Many of the characteristics that particularize the types of cheese are given by its natural mycobiota. The objective of this study was to determine the fungal species present in 20 samples of Serrano Artisanal Cheese from four ripening periods (14, 21, 28, and 35 days), produced in October 2017, in Santa Catarina. Identification of molds was made by identification keys and yeasts by MALDI-TOF. The molds were isolated in 28 of the 80 samples and with various species, some being attributed to contamination. On the other hand, yeasts were detected in all samples, with some genera being more frequent, specially Kluyveromyces lactis, present in 27 samples, and some species of Candida spp. The identified species have already been isolated in other Brazilian artisanal cheeses produced in places with similar climate and relief to that of Serrano Artisanal Cheese. (AU)

Micobiota do Queijo Artesanal Serrano produzido em Santa Catarina, Brasil / Mycobiota of Serrano Artisanal Cheese produced in Santa Catarina, Brazil

O Queijo Artesanal Serrano é um produto nativo do sul do Brasil, produzido por mão de obra familiar a partir do leite cru, e que vem buscando a obtenção da sua indicação geográfica. Muitas das características que particularizam os tipos de queijo são dadas pela sua micobiota natural. O objetivo deste estudo foi determinar as espécies fúngicas presentes em 20 amostras de Queijo Artesanal Serrano de quatro períodos de maturação (14, 21, 28 e 35 dias), produzidos em outubro de 2017, em Santa Catarina. A identificação dos bolores foi feita por chaves de identificação e das leveduras por MALDI-TOF. Os bolores foram isolados em 28 das 80 amostras e com espécies variáveis, sendo algumas contaminantes. Por outro lado, as leveduras foram detectadas em todas as amostras, com alguns gêneros mais frequentes, principalmente o Kluyveromyces lactis, presente em 27 amostras, e algumas espécies de Candida spp. As espécies identificadas já foram isoladas em outros queijos artesanais brasileiros, produzido em locais com características de clima e relevo semelhantes ao do Queijo Artesanal Serrano.

Serrano Artisanal Cheese is a native product from the South region of Brazil, produced by family labor from raw milk, which seeks to obtain its geographical indication. Many of the characteristics that particularize the types of cheese are given by its natural mycobiota. The objective of this study was to determine the fungal species present in 20 samples of Serrano Artisanal Cheese from four ripening periods (14, 21, 28, and 35 days), produced in October 2017, in Santa Catarina. Identification of molds was made by identification keys and yeasts by MALDI-TOF. The molds were isolated in 28 of the 80 samples and with various species, some being attributed to contamination. On the other hand, yeasts were detected in all samples, with some genera being more frequent, specially Kluyveromyces lactis, present in 27 samples, and some species of Candida spp. The identified species have already been isolated in other Brazilian artisanal cheeses produced in places with similar climate and relief to that of Serrano Artisanal Cheese.

Chemical diversity from the Tibetan Plateau fungi Penicillium kongii and P. brasilianum.

Two new secondary metabolites, kongiilines A and B (1, 7), and two asperphenamate derivatives, asperphenamates B and C (5-6), together with 16 known compounds (2-4, 8-20), were isolated from Tibetan Plateau fungi Penicillium kongii and Penicillium brasilianum. This is the first report on asperphenamates B and C as naturally occurring compounds, and that aspterric acid is isolated from P. brasilianum for the first time. Their structures were elucidated by different spectroscopic techniques including high-resolution electrospray ionisation mass spectrum, 1D nuclear magnetic resonance (NMR), and 2D NMR as well as electronic circular dichroism. Compounds 4, 5, and 10 exhibited cytotoxicity activities against human colon carcinoma HCT116 cell line with IC50 values of 88.16, 77.68, and 36.92 µM, respectively. Fungi from Tibetan Plateau represent important and rich resources for the investigation of new chemicals.

Isolation and identification of toxigenic and non-toxigenic fungi in samples of medicinal plants from the market / Isolamento e identificação de fungos toxigênicos e não toxigênicos em amostras plantas medicinais do comércio

ABSTRACT: The consumption of preparations of medicinal plants has been increasing during the last decades in occidental societies. The presence of toxigenic fungi in a plant product may represent a potential risk of contamination, because of aflatoxins and ochratoxins. In this study, 12 samples of medicinal plants were analyzed in relation to the level of fungal contamination, and the presence of producers of ochratoxin A and aflatoxins was assessed by visualization of fungi using a cromatovisor in coconut milk. Most of the species found belong to the genus Cladosporium, Fusarium, Aspergillus and Penicillium. Species producing ochratoxin A were present in 2 samples (16.7%), Melissa and Hibiscus. Species producing aflatoxin were found in samples of Jacaranda decurrens (8.33%). This study suggests that herbs, if stored improperly, can provide the growth of fungi and should be examined before consumption.

RESUMO: O consumo das plantas medicinais vem aumentando nas últimas décadas nas sociedades ocidentais, porém, a presença de fungos toxigênicos nestas plantas pode representar um risco em potencial de contaminação devido à produção de aflatoxinas e ocratoxinas. Neste trabalho, 12 amostras de plantas medicinais foram analisadas em relação ao nível de contaminação por fungos, enquanto a presença de produtores de ocratoxina A e aflatoxinas foi avaliada pela visualização em cromatovisor dos fungos em meio de leite de coco. A maioria das espécies encontradas pertence aos gêneros Cladosporium, Fusarium, Aspergillus e Penicillium. Espécies produtoras de ocratoxina A estavam presentes em 2 amostras (16,7%), Melissa e Hibisco. Espécies produtoras de aflatoxina foram encontradas na amostra de Carobinha (8,33%). Este trabalho sugere que as ervas, sendo armazenadas inadequadamente, proporcionam o crescimento de fungos e, por isso, estes devem ser examinados antes do consumo.

Una nueva generación de antifúngicos para el tratamiento de las micosis superficiales / A new generation of antifungals for the treatment of dermatomycosis

Las infecciones fúngicas superficiales responden bien a los tratamientos habituales en la mayoría de los casos pero, en determinadas situaciones, constituyen un problema. En su mayoría se trata de infecciones producidas por levaduras del género Candida y Malassezia y por hongos dermatofitos. Estas infecciones han visto incrementada su prevalencia junto con la selección de determinadas especies, la reducción de la sensibilidad a los antifúngicos y la aparición de fenómenos de resistencia in vitro e in vivo. La investigación para encontrar el antifúngico ideal aún continúa y en este sentido actualmente se están ensayando distintas estrategias de investigación sobre drogas para el tratamiento sistémico y tópico de las dermatomicosis.

Superficial fungal infections frequently caused by Candida spp. and Malassezia spp yeasts and dermatophytes fungi, have good response to common treatments in the majority of cases, but in some cases failure are described. Prevalence of these infections has been increased with the selection of certain species, reduced sensitivity to antifungal agents and the emergence of in vitro and in vivo resistance phenomena. The research to find the ideal antifungal still continues and in this sense are being currently tested different strategies for research on systemic and topical drugs for dermatomycosis treatment.

Non-dermatophyte mould onychomycosis: a clinical and epidemiological study at a dermatology referral centre in Bogota, Colombia.

Worldwide prevalence of non-dermatophyte mould onychomycosis has increased in recent years; however, available information on the topic is confusing and oftentimes contradictory, probably due to the small number of reported cases. The aim of this study was to determine and describe the aetiological agents, as well as the epidemiological and clinical characteristics of non-dermatophyte mould onychomycosis in a dermatology referral centre in Bogota, Colombia. A cross-sectional descriptive study was conducted between January 2001 and December 2011 among patients who attend the National Institute of Dermatology with a confirmed diagnosis of onychomycosis by non-dermatophytes moulds. There were 317 confirmed cases of non-dermatophyte mould onychomycosis in 196 women and 121 men whose average age was 43 years. Twenty-seven per cent of them had a history of systemic disease. The habit of walking and showering barefoot was the major infection-related factor. Distal and lateral subungual presentation was the most common pattern of clinical presentation. The most frequent non-dermatophyte mould was Neoscytalidium dimidiatum followed by Fusarium spp. No relationship was observed with predisposing factors previously reported in the literature. Clinical features found in this population are indistinguishable from onychomycosis caused by dermatophytes. High prevalence of N. dimidiatum found here was in contrast to a large number of studies where other types of moulds predominate.

Role of killer factors in the inhibitory activity of bio-control yeasts against Penicillium expansum and Aspergillus ochraceus

This work evaluated the antagonism of killer positive yeast strains (isolated from 11 samples of different frozen fruit pulps) against the strains of Penicillium expansum and Aspergillus ochraceus. Of the total 41 killer yeasts tested in YM agar, 19 showed antibiosis against P. expansum and A. ochraceus, with inhibition zone ranging from 10 to 18 mm and 10 to 19 mm, respectively. In the following step, the extracellular activity of Kluyveromyces sp. FP4(13) was tested performing the assay in YM broth. The antifungal activity of Kluyveromyces sp. FP4(13) cell-free culture supernatant (25ºC/96 h) was more effective against the conidia germination, showing inhibition rates of 93.33 and 86.44% for P. expansum and A. ochraceus, respectively. The micelial growth inhibition was 28.45 and 21.0%, respectively. The antagonism showed by the selected yeasts could be used as a promising alternative tool to reduce and control the postharvest fungal spoilage of the fruits. However, further studies should be carried out in order to better elucidate the role of innocuous characters in antagonistic microorganisms, as well as the purification and characterization of new killer toxins.

Mycotoxin in medicinal/aromatic Herbs: a review / Micotoxinas en hierbas medicinales y/o aromaticas: Revisión

Owing to their conditions of production, and sometimes to poor storage and transportation conditions, herbal products are susceptible to fungal contamination and development. This can lead to the accumulation of mycotoxins in this kind of commodities. Consequently, herbal products can be contaminated with fungal toxins, such as aflatoxins, ochratoxins and other mycotoxins, which pose a serious risk to public health. This paper reviews the main aspects regarding mycotoxin contamination of medical/aromatic herbs, in the context of the importance of this product in a global market. Moreover, the effect of processing on final contamination of derived foods, as well as the analytical methodology commonly employed in fungal and mycotoxin analysis in this type of products was reviewed.

Debido a sus condiciones habituales de producción, así como a condiciones inadecuadas de almacenamiento y de transporte, los productos a base de plantas medicinales o aromáticas son susceptibles a la contaminación por mohos, lo que puede conducir a la acumulación de toxinas fúngicas tales como las aflatoxinas, ocratoxinas y otras micotoxinas, lo que supone un grave riesgo para la salud pública. Este artículo revisa los principales aspectos de la contaminación por micotoxinas de las hierbas medicinales y/o aromáticas, teniendo en cuenta el contexto de este producto en un mercado globalizado, así como el efecto del procesado sobre la contaminación final de estos productos. También se revisa la metodología más comúnmente empleada en el análisis de mohos y micotoxinas en este tipo de productos.

Profilaxis antifúngica en niños y adultos sometidos a trasplante de órganos sólidos y de precursores hematopoyéticos / Prophylaxis against fungal infections in solid organ and hematopoietic stem cells transplantation

Invasive fungal infections are an important cause of morbidity and mortality in SOT and HSCT recipients. The main species involved are Candida spp. and Aspergillus spp, less frequently Cryptococcus spp., causal agents of mucormycosis and Fusarium spp. Usually occur within the first six months post-transplant, but they do it later, especially during episodes of rejection, which maintains the state of immune system involvement. Prophylaxis recommendations are specific to each type of transplant. In liver transplantation use of fluconazole is recommended only in selected cases by high risk factor for invasive fungal infections (A1). If the patient has a high risk of aspergillosis, there are some suggestions for adults population to use amphotericin B-deoxycholate, liposomal amphotericin B or caspofungin (C2) without being validated none of these recommendations in pediatric population. In adult lung transplant patients where the risk of aspergillosis is higher than in other locations, we recommend universal prophylaxis with itraconazole 200 mg/day, nebulised liposomal amphotericin B or voriconazole (C2), no validated recommendations for pediatrics. In HSCT, universal prophylaxis is recommended only in allogeneic and autologous selected cases. The most accepted indication is fluconazole (A1), and posaconazole (A1) or micafungin (A1) in selected cases with high risk of aspergillosis.

Las infecciones fúngicas invasoras constituyen una importante causa de morbilidad y mortalidad en los pacientes receptores de TOS y TPH. Los principales agentes involucrados son Candida spp. y Aspergillus spp, menos frecuentemente Cryptococcus spp., agentes causales de mucormicosis y Fusarium spp. Se presentan habitualmente dentro de los primeros seis meses posttrasplante, pero también lo hacen en forma más tardía, especialmente durante episodios de rechazo, en que se mantiene el estado de compromiso del sistema inmune. Existen recomendaciones de proilaxis especíicas para cada tipo de trasplante. En trasplante hepático se recomienda el uso de fluconazol sólo en casos seleccionados por factores de riesgo (A1). Si existe riesgo de asper-gilosis, hay algunas sugerencias en adultos para el uso de anfotericina B-deoxicolato, anfotericina liposomal o caspofungina (todo en categoría C2), sin estar validada ninguna de estas recomendaciones en pediatría. En trasplante pulmonar en paciente adulto, donde el riesgo de aspergilosis es superior a otras localizaciones, se recomienda proilaxis universal, con itraconazol 200 mg/día, anfotericina liposomal nebulizada o voriconazol (C2), sin recomendaciones validadas para pediatría. En TPH, se recomienda proilaxis universal en trasplante alogénico y sólo para casos seleccionados en trasplantes autólogos. La indicación más aceptada es fluconazol (A1), siendo alternativas a evaluar dependiendo del riesgo de aspergilosis, posaconazol (A1) y micafungina (A1).

Efeito in vitro do óleo essencial de Eucalyptus globulus sobre o crescimento e desenvolvimento de Penicillium sp / In vitro effect of Eucalyptus globulus essential oil on Penicillium sp

O fungo Penicillium sp. é o agente causal dos bolores, considerada a principal doença pós-colheita em citros, levando à perdas na qualidade e quantidade dos frutos comercializáveis. Além de perdas econômicas, uma vez que nas perdas pós-colheitas estão inclusos os custos, transporte e armazenagem de frutos. O presente trabalho teve por objetivo verificar a ação fungitóxica do óleo essencial de Eucalyptus globulus no controle in vitro de Penicillium sp., avaliando seu crescimento micelial, produção e a germinação de esporos. O experimento foi conduzido no laboratório de Microbiologia da PUC, campus Toledo, e constituiu-se de oito tratamentos, sendo: uso do óleo essencial de eucalipto nas concentrações de 1%, 0,5%, 0,25%, 0,1%, 0,05% e 0,025% adicionados ao meio de cultura BDA (Batata Dextrose-Ágar) autoclavado, testemunha negativa (meio de cultura com adição de 40 mg de ingrediente ativo azoxystrobin/L do fungicida) e testemunha positiva (apenas meio de cultura). O delineamento experimental utilizado foi inteiramente casualizado, com cinco repetições em cada tratamento. O óleo essencial de eucalipto inibiu o crescimento de forma significativa nas concentrações de 1%, 0,5% e 0,25%, não diferindo estatisticamente do controle com o fungicida azoxystrobin. Os tratamentos apresentaram o mesmo comportamento em relação a produção de esporos. Já na germinação de esporos, as concentrações de 1% e 0,5% obtiveram controle superior ao tratamento com azoxystrobin e a concentração de 0,25%. As demais concentrações de óleo não obtiveram resultados significativos nas avaliações, igualando-se estatisticamente a testemunha positiva (BDA).

Penicillium sp. is the casual agent of moulds, considered the main citrus post-harvest disease, causing loss on the quality and quantity of marketable fruits. Besides economic loss, it can be included costs, transportation and fruit storage problems. The present work had the objective to verify the fungitoxic action of Eucalyptus globules essential oil on the in vitro control of Penicillium sp., evaluating mycelial growth, production and spores germination. The experiment was conduced on the Microbiology laboratory of PUC, Toledo campus, with eight treatments: 1%, 0,5%, 0,25%, 0,1%, 0,05% e 0,025% concentrations of eucalyptus oil added to PDA media (potato-dextrose-agar), negative control (PDA with addition of 40mg of azoxystrobin/L) and positive control (PDA media only). The experimental design was according to a completely randomized design, with five replicates each treatment. The eucalyptus essential oil inhibited significantly the growth in the 1%, 0,5% e 0,25% concentrations, not differing statistically from control with fungicide azoxystrobin. The treatments presented the same behavior for spores production. For spores germination, the 1% e 0,5% concentrations had better control than treatment with azoxystrobin and the concentration of 0,25%. Others oil concentrations did not present significative results on the evaluations, being statistically equal to positive control (PDA).

In vitro effect of Eucalyptus globulus essential oil on Penicillium sp. / Efeito in vitro do óleo essencial de Eucalyptus globulus sobre o crescimento e desenvolvimento de Penicillium sp.

Penicillium sp. is the casual agent of moulds, considered the main citrus post-harvest disease, causing loss on the quality and quantity of marketable fruits. Besides economic loss, it can be included costs, transportation and fruit storage problems. The present work had the objective to verify the fungitoxic action of Eucalyptus globules essential oil on the in vitro control of Penicillium sp., evaluating mycelial growth, production and spores germination. The experiment was conduced on the Microbiology laboratory of PUC, Toledo campus, with eight treatments: 1%, 0,5%, 0,25%, 0,1%, 0,05% e 0,025% concentrations of eucalyptus oil added to PDA media (potato-dextrose-agar), negative control (PDA with addition of 40mg of azoxystrobin/L) and positive control (PDA media only). The experimental design was according to a completely randomized design, with five replicates each treatment. The eucalyptus essential oil inhibited significantly the growth in the 1%, 0,5% e 0,25% concentrations, not differing statistically from control with fungicide azoxystrobin. The treatments presented the same behavior for spores production. For spores germination, the 1% e 0,5% concentrations had better control than treatment with azoxystrobin and the concentration of 0,25%. Others oil concentrations did not present significative results on the evaluations, being statistically equal to positive control

O fungo Penicillium sp. é o agente causal dos bolores, considerada a principal doença pós-colheita em citros, levando à perdas na qualidade e quantidade dos frutos comercializáveis. Além de perdas econômicas, uma vez que nas perdas pós-colheitas estão inclusos os custos, transporte e armazenagem de frutos. O presente trabalho teve por objetivo verificar a ação fungitóxica do óleo essencial de Eucalyptus globulus no controle in vitro de Penicillium sp., avaliando seu crescimento micelial, produção e a germinação de esporos. O experimento foi conduzido no laboratório de Microbiologia da PUC, campus Toledo, e constituiu-se de oito tratamentos, sendo: uso do óleo essencial de eucalipto nas concentrações de 1%, 0,5%, 0,25%, 0,1%, 0,05% e 0,025% adicionados ao meio de cultura BDA (Batata Dextrose-Ágar) autoclavado, testemunha negativa (meio de cultura com adição de 40 mg de ingrediente ativo azoxystrobin/L do fungicida) e testemunha positiva (apenas meio de cultura). O delineamento experimental utilizado foi inteiramente casualizado, com cinco repetições em cada tratamento. O óleo essencial de eucalipto inibiu o crescimento de forma significativa nas concentrações de 1%, 0,5% e 0,25%, não diferindo estatisticamente do controle com o fungicida azoxystrobin. Os tratamentos apresentaram o mesmo comportamento em relação a produção de esporos. Já na germinação de esporos, as concentrações

Avaliação da qualidade microbiológica do mel comercializado no extremo sul da Bahia / Evaluation of the microbiological quality of honey commercialized in Bahia extreme south region

Este estudo teve como objetivo analisar a qualidade microbiológica do mel comercializado formal e informalmente na região do extremo sul baiano, quanto à presença de coliformes totais, termotolerantes, bolores e leveduras. Foram analisadas amostras de mel adquiridas no comércio formal e informal. As análises microbiológicas das amostras foram de enumeração de coliformes totais e termotolerantes, por meio do Número Mais Provável (NMP) e contagem de bolores e leveduras. Foram analisadas 18 amostras, sendo dez adquiridas no comércio formal e oito no informal. Todas estavam isentas de contaminação por bactérias do grupo coliformes, tanto totais como termotolerantes, no entanto oito (44,44%) - quatro do comércio formal e quatro do informal - continham bolores e leveduras. De acordo com as características gerais das amostras de mel analisadas neste estudo apenas quatro (22,22%) encontravam-se próprias para o consumo, segundo os padrões exigidos pelo Regulamento Técnico para Fixação e Identidade e Qualidade do Mel.

The objective of this study is to analyze the microbiological quality of honey commercialized in Bahia's extreme south region, in order to search for the presence of total and thermo tolerant coliforms, moulds and yeasts. Honey samples, eight of which acquired in the formal commerce and ten obtained in the informal commerce were analyzed. The microbiological analyses of samples consisted of enumerating total and thermo tolerant coliforms through the Most Probable Number (MPN) and the counting of moulds and yeasts. All samples were contamination free for coliforms. However, eight (44,44%) of the 18 (100%) samples obtained in both formal and informal commerce contained moulds and yeasts. In accordance to the general characteristics of the samples of honey analyzed in this study, only four (22,22%) met proper conditions for consumption.

Actualización en pruebas de susceptibilidad antifúngica / An update on antifungal susceptibility testing

Due to increasing of invasive fungal infections and emergeney of antifungal drugs resistant fungi, standardized methods of antifungal susceptibility testing (AST) have been developed. The Clinical Laboratory Standards Instutute (CLSI) and the European for Committee Antimicrobial Susceptibility Testing (EUCAST) have guidelines for susceptibility of yeasts by broth microdilution (M27-A2 and E. Dis. 7.1 documents, respectively). Both are equivalent, although they present methodological and interpretative breakpoints differences. In addition, the CLSI have the M38-A (for filamentous fungi) and M44-A (disk diffusion) documents, whereas EUCAST is developing a document for Aspergillus spp. Furthermore, commercial methods are available that display good correlation with the methods of reference such as E-test®, Sensititre® and Vitek2®. The interpretation of the results must be careful because the determination of the minimum inhibitory concentration (CIM) is difficult for fungi, there are host factors involved and not always there is a correlation between MIC and clinical outeome. Due to these methods are laborious and require trained personnel, to ask for AST to a reference laboratory is recommendable.

Debido al aumento en las infecciones fúngicas invasores y a la emergencia de hongos resistentes a los antifúngicos, ha sido necesario desarrollar métodos estandarizados de susceptibilidad antifúngica. El Clinical Laboratory Standards Instutute (CLSI) y el European Committee for Antimicrobial Susceptibility Testing (EUCAST) han elaborado guías para susceptibilidad de levaduras por microdilución en caldo (documentos M27-A2 y E. Dis. 7.1, respectivamente). Ambos son equivalentes, aunque presentan diferencias metodológicas y en sus puntos de corte. El CLSI ha desarrollado los documentos M38-A (hongos filamentosos) y M44-A (difusión en disco), mientras que EUCAST trabaja en un documento para Aspergillus sp. Por otra parte, existen métodos comerciales que presentan buena correlación con los métodos de referencia como E-test®, Sensititre® y Vitek2®. La interpretación de los resultados debe ser cuidadosa pues la determinación de la concentración inhibitoria mínima (CIM) es muy dificultosa para hongos, hay factores del hospedero involucrados y no siempre hay una correlación entre la CIM y la respuesta a tratamiento. Como estas técnicas, en general, son laboriosas y requieren de personal entrenado, es recomendable derivar los estudios de susceptibilidad a un laboratorio de referencia.

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 percent 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 percent. In our prospective study, in which we had data from successive mycological examinations, 37.8 percent 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.

Inhibitory effect of Cinnamomum zeylanicum Blume (Lauraceae) essential oil and beta-pinene on the growth of dematiaceous moulds

Dematiaceous moulds are pathogen microorganisms able to act as etiological agents of mycoses with different degrees of severity. This study evaluated the effectiveness of Cinnamomum zeylanicum Blume essential oil and beta-pinene in inhibiting the growth of various strains of dematiaceous moulds (Alternaria brassicola, Cladosporium herbarium, C. resinae, C. cladosporioides, Chaetomum globosum, Curvularia sp., Fonsecaea compacta, Piedraia hortae). Antimicrobial assays were led by determining the Minimum Inhibitory Concentration-MIC using the solid medium diffusion procedure and observing the interference of the MIC values on the mould radial mycelial growth along 14 days. MIC values found to C. zeylanicum essential oil oscillated between 63 and 125 µL/mL. beta-pinene showed MIC value of 125 µL/mL for the most mould strains, however C. resinae and C. globosum were resistant to it in all assayed concentrations. MIC values found to C. zeylanicum essential oil and beta-pinene presented intense fungicidal effect noted by a total inhibition of the mycelial growth of C. cladosporioides and F. compacta along 14 days of exposure. These results showed the intense antimould potential of C. zeylanicum essential oil and beta-pinene which could be regarded in a rational use in pharmaceutical formulations used to treat some mycoses, particularly, those caused by dematiaceous moulds.

Os fungos dematiáceos são microrganismos patogênicos capazes de agir como agentes etiológicos de micoses com diferentes graus de severidade. Este estudo avaliou a efetividade do óleo essencial de Cinnamomum zeylanicum Blume e beta-pineno em inibir o crescimento de várias cepas de fungos dematiáceos (Alternaria brassicola, Cladosporium herbarium, C. resinae, C. cladosporioides, Chaetomum globosum, Curvularia, Fonsecaea compacta, Piedraia hortae). Os ensaios antimicrobianos foram conduzidos através da determinação da Concentração Inibitória Mínima-CIM utilizando-se da técnica de difusão em meio sólido, bem como através da observação da interferência dos valores de CIM sobre o crescimento micelial radial fúngico ao longo de 14 dias. Os valores de CIM encontrados para o óleo essencial de C. zeylanicum oscilaram entre 63 e 125 µL/mL. beta-pineno apresentou um valor de CIM de 125 µL/mL para a maioria das cepas fúngicas, entretanto C. resinae e C. globosum mostraram-se resistentes a todas as concentrações ensaiadas. Os valores de CIM encontrados para C. zeylanicum e beta-pineno apresentaram intenso efeito fungicida notado por uma total inibição do crescimento micelial de C. cladosporioides e F. compacta ao longo de 14 dias de exposição. Estes resultados mostraram o intenso potencial antifúngico do óleo essencial de C. zeylanicum e beta-pineno, os quais poderiam ser considerados em um uso racional em formulações farmacêuticas utilizadas para a terapia de algumas micoses, particularmente, aquelas causadas por fungos dematiáceos.