Desenvolvimento de metodologia para identificação de fungos através de metabarcoding aplicada em amostras humanas de adenocarcinoma gástrico e carcinoma epidermoide de pênis / Development of methodology for fungi identification through metabarcoding applied in gastric adenocarcinoma and penile epidermoid carcinoma of human sample

O microbioma humano compreende material genético da microbiota de um local do corpo e tem influência direta ou indireta na manutenção da homeostase. O distúrbio da microbiota pode estar relacionado ao desenvolvimento de doenças. A população fúngica ainda é muito pouco estudada no contexto do microbioma. No presente estudo, foi desenvolvida uma metodologia para identificação de fungos por metabarcoding. A metodologia desenvolvida foi aplicada em mostras de pacientes portadores de adenocarcinoma gástrico ou carcinoma epidermoide de pênis. De modo geral, em ambos os tumores foi verificada a redução de diversidade fúngica conforme a evolução do estadiamento patológico. Também foram verificados resultados não concordantes ao analisar espécies diferencialmente abundantes em dados de sequenciamento da região ITS2 e de WGS nas amostras de lavado gástrico. Este trabalho reforça a importância em se estudar os fungos e sua associação com doenças como o câncer e incentiva próximos estudos através do desenvolvimento de uma metodologia específica para o micobioma.

The human microbiome comprises genetic material from the microbiota of a body site and has a direct or indirect influence on the maintenance of homeostasis. The disturbance of the microbiota may be related to the development of diseases. The fungal population is still very little studied in the context of the microbiome. In this study, a methodology was developed to identify fungi by metabarcoding. The methodology developed was applied to samples from patients with gastric adenocarcinoma or squamous cell carcinoma of the penis. In general, in both tumors, a reduction in fungal diversity was observed according to the evolution of the pathological staging. Discordant results were also found when analyzing differentially abundant species in sequencing data from the ITS2 region and WGS in gastric lavage samples. This work reinforces the importance of studying fungi and their association with diseases such as cancer and encourages further studies through the development of a specific methodology for the mycobiome

Correlation analysis between continuous cropping obstacle of Gastrodia elata and Ilyonectria fungi and relieving strategy / 中国中药杂志

The continuous cropping obstacle of Gastrodia elata is outstanding, but its mechanism is still unclear. In this study, microbial changes in soils after G. elata planting were investigated to explore the mechanism correlated with continuous cropping obstacle. The changes of species and abundance of fungi and bacteria in soils planted with G. elata after 1, 2, and 3 years were compared. The pathogenic fungi that might cause continuous cropping diseases of G. elata were isolated. Finally, the prevention and control measures of soil-borne fungal diseases of G. elata were investigated with the rotation planting pattern of "G. elata-Phallus impudicus". The results showed that G. elata planting resulted in the decrease in bacterial and fungal community stability and the increase in harmful fungus species and abundance in soils. This change was most obvious in the second year after G. elata planting, and the soil microbial community structure could not return to the normal level even if it was left idle for another two years. After G. elata planting in soils, the most significant change was observed in Ilyonectria cyclaminicola. The richness of the Ilyonectria fungus in soils was significantly positively correlated with the incidence of G. elata diseases. When I. cyclaminicola was inoculated in the sterile soil, the rot rate of G. elata was also significantly increased. After planting one crop of G. elata and one to three crops of P. impudicus, the fungus community structure in soils gradually recovered, and the abundance of I. cyclaminicola decreased year by year. Furthermore, the disease rate of G. elata decreased. The results showed that the cultivation of G. elata made the Ilyonectria fungi the dominant flora in soils, and I. cyclaminicola served as the main pathogen of continuous cropping diseases of G. elata, which could be reduced by rotation planting with P. impudicus.

Aflatoxins and fungal community distribution during harvesting and processing of Platycladi Semen / 中国中药杂志

This study aimed to analyze aflatoxins content and fungal community distribution in the harvesting and processing of Platycladi Semen, and explore the key link that affects aflatoxins contamination. The related Platycladi Semen samples of different maturity periods(cone non-rupture period, early rupture, and complete rupture period) and different processing periods(before drying, during 2-d drying, during 7-d drying, before and after seed scale removal, before and after peeling, 1 d after color sorting, and 7 d after color sorting) were collected for identifying the fungal community composition on sample surface by ITS amplicon sequencing. Then the content of aflatoxins B_1, B_2, G_1 and G_2 was determined by HPLC-MS/MS. The results showed that during the harvesting of Platycladi Semen from cone non-rupture to complete rupture, aflatoxins were only detected in the seed scale and seed coat, with aflatoxin G_2 in the seed scale and aflatoxin B_1 in the seed coat. During the drying, with the prolongation of drying time, aflatoxins B_1 and G_2 were detected simultaneously in the seed scale, aflatoxin B_1 in the seed coat, and low-content aflatoxin B_1 in the seed kernel. During subsequent processing, the aflatoxin content in seed kernel during subsequent processing was slighted increased. As demonstrated by fungal detection, Aspergillus flavus was not present during the harvesting of Platycladi Semen, but present during the drying and processing. Its content in the seed coat during the drying process was relatively higher. In short, Platycladi Semen should be harvested as soon as possible after it becomes fully mature. Drying process is the key link of preventing aflatoxin contamination. It is advised to build a sunlight room or adopt similar settings, standardize the operations in other processes, and keep the surrounding environment clean to minimize aflatoxin contamination.

Effects of glyphosate on soil fungal communities: A field study / Efecto del glifosato en las comunidades fúngicas del suelo: estudio a campo

ABSTRACT The driving forces behind many soil processes are microorganisms and they are able to respond immediately to environmental changes. The soil microbial community impacts on many soil properties. More than one-third of the terrestrial ecosystems are semiarid. However, a limited number of studies have been conducted to characterize soil fungal communities in semiarid grasslands, in particular those of agricultural fields. The aim of this study was to explore changes in the diversity and structure of soil fungal communities in semiarid grasslands, after different doses of glyphosate were applied under field conditions. Changes in soil fungal communities were examined using different approaches including culturing, calcofluor white stain and denaturing gradient gel electrophoresis (DGGE). The different approaches complement each other, revealing different aspects of the effect of glyphosate on soil fungal communities. We demonstrated a negative effect of glyphosate on soil fungal biomass at high doses and an early and transitory stimulatory effect on soil fungal biomass. We also found a negative effect of glyphosate on the species richness of cultivable fungi and changes in the molecular structure of soil fungal communities after double doses or long-term glyphosate application. In summary, our findings demonstrate an overall negative effect of glyphosate on soil fungal communities.

RESUMEN Los microorganismos del suelo son los responsables de llevar a cabo la mayoría de los procesos biológicos que ocurren en el suelo, y son capaces de reaccionar ante el estrés ambiental. Más de un tercio de los ecosistemas terrestres son semiáridos. Sin embargo, son escasos los estudios realizados para caracterizar las comunidades fúngicas en suelos agrícolas en ecosistemas semiáridos. El objetivo del presente trabajo fue estudiar los cambios que se producen en la biomasa, la diversidad y la estructura de las comunidades fúngicas del suelo, luego de la aplicación de distintas dosis de glifosato en condiciones de campo. Se emplearon diferentes técnicas incluidas el cultivo, la tinción directa con blanco de calcoflúor y PCR acoplada a electroforesis en geles de gradiente desnaturalizante (DGGE). Las distintas metodologías empleadas se complementan entre sí al detectar cada una distintos aspectos del efecto del glifosato en las comunidades fúngicas del suelo. Se encontró que el glifosato produce un efecto negativo sobre la biomasa fúngica, también se encontró un efecto transitorio estimulante inmediatamente posterior a la aplicación del herbicida. Además, se vio un efecto negativo sobre la riqueza de hongos cultivables, así como también cambios en la estructura molecular de las comunidades luego de aplicaciones repetidas. En conclusión, se demostró un efecto negativo generalizado sobre las comunidades fúngicas del suelo.

Investigation and research on fungal diversity of Platycladi Semen on Chinese market / 中国中药杂志

In the process of harvesting, production and processing, storage, and transportation, the traditional Chinese medicine Platycladi Semen is prone to mildew due to its own and environmental factors, which can nourish the production of toxic or pathogenic fungi, and even produce mycotoxins, which affects the safety of clinical medication. The 2020 edition of Chinese Pharmacopoeia limits the highest standard of aflatoxin content in Platycladi Semen. However, there are few studies on the fungal contamination of Platycladi Semen, and it is difficult to prevent and control it in a targeted manner. Therefore, based on the Illumina NovaSeq6000 platform, this article uses ITS sequence amplicon technology to analyze the distribution and diversity of fungi in 27 batches of commercially available Platycladi Semen in the Chinese market. A total of 10 phyla, 35 classes, 93 orders, 193 families, 336 genera, and 372 species of fungi were identified in China. Among them, Aspergillus, Alternaria spp. were dominant, 20 batches of samples were detected for A. flavus, 10 batches of samples were detected for A. nidulans, and all samples were detected for potential pathogenic fungi such as A. fumigatus and A. niger. According to diversity analysis, the diversity of the fungal communities in the samples from Gansu province was high, the samples in Shandong province contain the largest number of fungal species, and the samples in Guangxi province had the lo-west diversity and the least number of species. In most samples, pathogenic fungi such as A. fumigatus, A. niger, A. flavus, A. parasiticus were detected in varying degrees. This study systematically investigated the fungal contamination of Platycladi Semen from the markets in the last link of the its industrial chain, and clarified the distribution of Platycladi Semen fungi, especially toxin-producing fungi, and provided theoretical basis for the targeted prevention and control of fungal contamination in Platycladi Semen.

Antimicrobial activity in cultures of endophytic fungi isolated from Talinum triangulare

Endophytic fungi colonize the interior of plants without causing damage and act in symbiosis with their host. They are also potential sources of compounds with potential applications in industry, agriculture, and medicine, Thus, this study aimed to isolate and identify the endophytic fungi medicinal plant Talinum triangulare and evaluate its potential for the production of antimicrobial substances using the disk diffusion technique and testing in liquid medium front of Staphylococcus aureus ATCC SA 6538, Pseudomonas aeruginosa ATCC PA 0030, and Corynebacterium diphtheria ATCC 27012. Corynebacterium diphtheria was isolated from 3 fungi of the genus Trichoderma and Penicillium, with only the genus Trichoderma fungi showing antimicrobial activity.

Variation in fungal community structures in rhizosphere soil of Coptis chinensis with cropping mode under natural forest and artificial shed / 中国中药杂志

This study is aimed to reveal the rhizosphere soil fungal community structure difference of Coptis chinensis cropping between natural forest and artificial shed modes, and provide theoretical guidance for soil improvement and C. chinensis planting. The rhizosphere soil samples of 1-5-year-old C. chinensis under natural forest and artificial shed modes were collected. Illumina high-throughput sequencing technology was used to analyze the community structure and diversity of soil fungi under the tow cropping modes,and the effects of soil nutriment indices on soil fungal community structure. The results suggested that the abundance and diversity of fungal communities in soil of 2-5-year-old C. chinensis were not significant different in both two cropping modes, but it was significantly higher than that in the 1-year-old C. chinensis. Comparing soil samples from the same year-old C. chinensis under the two cropping modes, it was found that there was no significant difference in the abundance and diversity of fungal communities. The fungal community of the rhizosphere soil was different in composition and abundance between tow cropping modes, and between different planting years. The 17 phyla,59 classes and 155 orders,and 17 phyla,59 classes and 157 orders were detected in the rhizosphere soil of C. chinensis under the cropping modes of natural forest and the artificial shed, respectively. Ascomycota, Basidiomycota and Mortierellomycota were dominant phyla in rhizosphere soil, and the average abundance of the 3 phyla accounted for 74.36% and 74.30% of the total fungi. The results of analysis of similarities showed that there were significant differences in the fungal community structure of 1-year-old and 2-year-old C. chinensis soil fungi, and there was no significant difference in the community structure of 3-5-year-old samples. Under the natural forest cropping mode, there were significant differences among the samples of different years. Under the artificial shed cropping, there were significant differences in fungal community structure between 1-year-old and 3-5-year-old C. chinensis soil, and between 2-year-old and 3-5-year-old C. chinensis soil. The results of canonical correlation analysis showed that soil pH and soil organic matter content were the main factors affecting the soil fungal community structure. Soil organic matter content was positively correlated with Basidiomycota and Cryptomycota, pH was negatively correlated with Basidiomycota and C. ryptomycota. The planting of C. chinensis has promoted the diversity and abundance of rhizosphere fungal community significantly. For the same year-old C. chinensis soil, abundance of fungal community was no significant difference between two cropping modes. There are significant differences in the rhizosphere soil fungal community structure between tow cropping modes in the first two years of planting. Through the interaction between the rhizosphere and the soil and the continuous selection of the rhizosphere to the fungal community, the fungal community structure tended to be the same between the two cropping modes in rhizosphere soil of 3-5-year old C. chinensis. The soil pH and orga-nic matter content were the main factors affecting the change of fungal community structure.

Yeast diversity in Vitis non-vinifera ecosystems / Diversidad de levaduras en ecosistemas de Vitis no-vinifera

The surface of grapes lodges a complex community of yeast species responsible for spontaneous alcoholic fermentation. The study of indigenous Saccharomyces and "non-Saccharomyces" yeasts during grape must fermentation constitutes a major research area in microbial enology. Although there are detailed studies on the microbiota of Vitis vinifera grapes, little is known about the diversity of yeast communities present in non-vinifera Vitis ecosystems (i.e., grapes and spontaneously fermenting grape musts). Potentially scientific and/or enological valuable yeast strains from these non-vinifera Vitis ecosystems might never be isolated from V. vinifera L. In this updated review, we summarize relevant aspects of the microbial studies conducted on V. non-vinifera grapes and spontaneously fermenting grape musts.

La superficie de las uvas aloja una comunidad compleja de especies de levaduras responsables de la fermentación alcohólica espontánea. El estudio de estas levaduras Saccharomyces y «no-Saccharomyces¼ durante la fermentación del mosto de uvas constituye un área relevante de investigación microbiológica en enología. Si bien existen estudios detallados de la microbiota de uvas de Vitis vinifera L., poco se sabe sobre la diversidad de comunidades de levaduras presentes en ecosistemas de Vitis no-vinifera (i.e., uvas y mostos en fermentación espontánea). Cepas de levaduras presentes en ecosistemas de Vitis no-vinífera, con valor potencial científico y/o enológico, podrían no estar presentes en V. vinifera L. En esta revisión actualizada, resumimos los aspectos relevantes de los estudios microbiológicos efectuados en mostos en fermentación espontánea de uvas de V. no-vinifera.

Cutaneous mycobiota of boid snakes kept in captivity / Microbiota cutânea de serpentes Boidae mantidas em cativeiro

Boids are large, constrictor snakes that feed mostly on mammals, reptiles, and birds. These animals are commonly raised as pets, and their improper handling can favor the emergence of fungal infections, which can lead to dermatological diseases that are undiagnosed in nature. Here, we isolate and identify the filamentous fungi that compose the mycobiota of the scales of boid snakes kept in captivity at the Biological Museum of the Butantan Institute. Thirty individuals of four species were evaluated: four Eunectes murinus, twelve Boa constrictor constrictor, seven Corallus hortulanus, and seven Epicrates crassus. Microbiological samples were collected by rubbing small square carpets on the snake scales. We isolated five genera of fungi: Penicillium sp. (30%), Aspergillus sp. (25%), Mucor sp. (25%), Acremonium sp. (10%), and Scopulariopsis sp. (10%). Approximately half of the snakes evaluated had filamentous fungi on the scales, but only 12% of the individuals were colonized by more than one fungal genus. We found no dermatophytes in the evaluated species. Our results provide an overview of the fungal mycobiota of the population of boids kept in the Biological Museum, allowing the identification of possible pathogens.(AU)

Boidae são serpentes grandes e constritoras, que comem normalmente mamíferos, répteis e aves. São comumente criadas como animais de estimação, e seu manuseio inadequado pode favorecer infecções fúngicas emergentes, as quais podem causar doenças dermatológicas que não são diagnosticadas na natureza. Foram isolados e identificados os fungos filamentosos da microbiota das escamas das serpentes Boidae mantidas em cativeiro no Museu Biológico do Instituto Butantan. Trinta indivíduos de quatro espécies foram avaliados: quatro Eunectes murinus, 12 Boa constrictor constrictor, sete Corallus hortulanus e sete Epicrates crassus. As amostras microbiológicas foram coletadas por fricção, em pequenos quadrados de carpete das escamas das serpentes. Isolaram-se cinco gêneros de fungos: Penicillium sp. (30%), Aspergillus sp. (25%), Mucor sp. (25%), Acremonium sp. (10%) e Scopulariopsis sp. (10%). Aproximadamente metade das serpentes avaliadas tinham fungos filamentosos nas escamas e apenas 12% dos indivíduos foram colonizados por mais de um gênero. Não foram encontrados dermatófitos nas espécies avaliadas. Os resultados mostraram uma visão geral da microbiota fúngica da população de boídeos mantidos no Museu Biológico, o que permitiu a identificação de possíveis patógenos.(AU)

Colonización por hongos ambientales en el paciente alérgico respiratorio no controlado / Colonization by environmental fungi in the uncontrolled respiratory allergic patient

Introducción: La presencia de hongos en las vías respiratorias puede provocar en personas susceptibles diversas manifestaciones alérgicas. Objetivo: Determinar si las especies fúngicas aisladas de la mucosa nasal de pacientes alérgicos respiratorios pueden ser definidas como alergenos sensibilizantes a través de las pruebas cutáneas. Métodos: Estudio observacional, prospectivo y de corte transversal, donde el universo estuvo constituido por todos los pacientes con diagnóstico de alergia respiratoria o inicio de asma bronquial variable descompensada, mayores de 2 años y menores de19,que se asistieron en las consultas de alergia en La Habana, desde enero 2016 a enero 2017. La muestra obtenida fue de 80 pacientes alérgicos respiratorios. Resultados: Del total de pacientes con clínica de asma, rinitis o ambas, se obtuvieron pruebas cutáneas por el test de Prick positivas a hongos en 52 de ellos (65 por ciento) con una polisensibilización en 24 para 46,1 por ciento. La reactividad cruzada de mayor relevancia se produjo con los alérgenos de Aspergillus, Penicillium y Alternaria. El cultivo resultó positivo en 54 muestras nasales (67 por ciento). El género de hongos predominante en los pacientes alérgicos fue el Aspergillus en 70,3 por ciento y dentro de este el Aspergillus fumigatus en 52,6 por ciento. Conclusiones: El estudio de la micobiota nasal es una prueba que debe interpretarse junto con las pruebas cutáneas para el diagnóstico de enfermedades alérgicas por hongos ambientales y tener en cuenta su importancia para el control epidemiológico en la exposición a hongos(AU)

Introduction: The presence of fungi in the respiratory tract can cause different allergic manifestations in sensitive persons. Objective: To determine if fungi species isolated from the nasal mucosa of respiratory allergic patients can be defined as allergen-sensitive by means of skin tests. Methods: Observational, prospective and cross-sectional study, where the overall sample consisted of all patients with a diagnosis of respiratory allergy or onset of decompensated variable bronchial asthma, over 2 years old and under 19 years old who attended to Allergy consultations in Havana, from January 2016 to January 2017. The sample obtained was 80 respiratory allergic patients. Results: Of the total number of patients with symptoms of clinical asthma, rhinitis or both, Prick´s skin tests were obtained by fungal positive test in 52 of them (65 percent) with a polysensitization in 24 (46.1 percent). The most relevant cross reactivity occurred with the allergens of Aspergillus, Penicillium and Alternaria. The culture was positive in 54 nasal samples (67 percent). The predominant fungal genus in allergic patients was Aspergillus in 70.3 percent and within this Aspergillus fumigatus in 52.6 percent. Conclusions: The study of nasal mycobiota is a test that should be interpreted together with skin tests for the diagnosis of allergic diseases due to environmental fungi and it must be taken into account its importance for epidemiological control in fungal exposure(AU)

Human mycobiome and diseases / 华西口腔医学杂志

The proportion of mycobiome is less than 1% of human microbiome. However, fungal community plays a key role in human health and diseases. With high-throughput sequencing applications, the structure and composition of mycobiome in the mouth, lung, gut, vagina, and skin have been analyzed, and the role of microbiome in diseases has been investigated. Mycobiome also influences the composition of bacteriome and includes key species that maintain the structure and function of microbial communities. Fungi also influence host immune responses. In this review, we summarized the mycobiome com-position at various sites and different diseases and the interactions between fungi-bacteria and fungi-host.

Salivary mycobiome dysbiosis and its potential impact on bacteriome shifts and host immunity in oral lichen planus / 国际口腔科学杂志·英文版

The biodiversity of the mycobiome, an important component of the oral microbial community, and the roles of fungal-bacterial and fungal-immune system interactions in the pathogenesis of oral lichen planus (OLP) remain largely uncharacterized. In this study, we sequenced the salivary mycobiome and bacteriome associated with OLP. First, we described the dysbiosis of the microbiome in OLP patients, which exhibits lower levels of fungi and higher levels of bacteria. Significantly higher abundances of the fungi Candida and Aspergillus in patients with reticular OLP and of Alternaria and Sclerotiniaceae_unidentified in patients with erosive OLP were observed compared to the healthy controls. Aspergillus was identified as an "OLP-associated" fungus because of its detection at a higher frequency than in the healthy controls. Second, the co-occurrence patterns of the salivary mycobiome-bacteriome demonstrated negative associations between specific fungal and bacterial taxa identified in the healthy controls, which diminished in the reticular OLP group and even became positive in the erosive OLP group. Moreover, the oral cavities of OLP patients were colonized by dysbiotic oral flora with lower ecological network complexity and decreased fungal-Firmicutes and increased fungal-Bacteroidetes sub-networks. Third, several keystone fungal genera (Bovista, Erysiphe, Psathyrella, etc.) demonstrated significant correlations with clinical scores and IL-17 levels. Thus, we established that fungal dysbiosis is associated with the aggravation of OLP. Fungal dysbiosis could alter the salivary bacteriome or may reflect a direct effect of host immunity, which participates in OLP pathogenesis.

Isolation and identification of molds flora in external genital tract of healthy mares

Background: Microorganisms are responsible for development of many diseases, which directly or indirectly affect reproductive performance in mares, like various animal species. Fungi are present at mucous membranes of reproductive tract as mycoflora in warm blooded animals

Objectives: The aim of this study was isolation and identification of molds of external reproductive tract in mares

Methods: Samples were collected from genital tract of 151 mares in different ages from horse breeding farms and clubs suburb of Tehran from April to December, 2014. Age, pregnancy, insemination and mating were recorded. Samples were taken by sterile cotton swabs from 3 different sites, [1] vestibule, [2] clitoral fossa and [3] vagina and transferred under cold condition to the laboratory of Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran,Iran. Samples were inoculated in Sabouraud glucose agar supplemented with antibiotic [chlor-amphenicol; 0.005] and kept at 30°C for 10-15 days before being considered negative. Fungal identification relied on morphologic and physiologic features. SPSS Version 20 was used for statistical analysis of the data

Results: Totally, 666 filamentous fungi colonies were isolated. Maximum colony [CFUs] were 20 and 37 colonies for each site and horse, respectively. In total, 81.5 percent of mares and 48.3 percent of different sites were positive. The most predominant isolates were Aspergillus spp 362[54.35%], Scopulariop-sis 93[13.96%], Cladosporium 35[5.25%], Penicillium 31[4.65%], Alternaria 26[3.90%], Fusarium 20[3.03%]. Aspergillus took high occurrence with 362 and A. fumigatus with 156 colonies followed by Scopulariopsis 93, Cladosporium 35 and Pencillium 31. A. fumigatus had significant difference among other fungi p<0.05. Among sites, 39.51% for filamentous fungi and 8.17% for both filamentous fungi and yeasts were positive.265

Conclusions: Isolated filamentous fungi could be resident flora of external reproductive tract of mares. These saprophytic pathogen fungi in predisposing conditions could be causative agents of endometritis and infertility in mares

Hongos productores de micotoxinas asociados a hierbas medicinales en la Ciudad de São Paulo, Brasil / Mould and mycotoxins associated in medicinal plants in São Paulo, Brazil

Antecedentes: la fitoterapia es una de las más antiguas prácticas utilizadas por la humanidad. Hasta mediados del siglo XIX, cuando se introdujeron los medicamentos, la formulación de estos generalmente era basada en plantas medicinales. Objetivos: Determinar la micobiota y los niveles de aflatoxinas originadas de Aspergillus sección Flavi aislados de las 50 muestras de medicamentos fitoterápicos comercializados actualmente en la ciudad de São Paulo, Brasil. Métodos: Cincuenta (50) muestras de medicamentos fitoterápicos en la forma de hojas (té-25) y cápsulas (25) fueron colectadas de agosto de 2000 a julio de 2001. Los hongos filamentosos aislados fueron identificados al nivel de género de acuerdo con las características morfológicas y criterios taxonómicos. El análisis de aflatoxinas fue realizada por cromatografía de capa fina (TLC). Resultados: El análisis microbiológico mostró que 41 (82 por ciento) de los medicamentos fitoterápicos presentaron un crecimiento fúngico sobre las 100 UFC/g. Un total de 106 especies de seis diferentes géneros fueron aislados (Aspergillus, Penicillium, Mucor, Rhizopus y Alternaria). El género Aspergillus fue el predominante (60.5 por ciento) seguido por Penicillium (20,0 por ciento). Aspergillus niger (30 por ciento) A. flavus (22 por ciento), A. fumigatus (6,5 por ciento) y A. parasiticus fueron las especies de Aspergillus identificadas. Se observó que 13 (56,5 por ciento), de los 23 A. flavus aislados y dos aislados de A. parasiticus produjeron aflatoxinas. Conclusiones: La contaminación observada en la mayoría de los productos y el alto nivel de cepas productoras de aflatoxinas justifica un análisis más cuidadoso de los medicamentos fitoterápicos comercializados y la aplicación de leyes más rigurosas son necesarias para garantizar la calidad de los productos.

Background: phytotherapy is one of the most ancient practices used by humanity. In Antiquity until the middle of the XIX century, when chemotherapeutic drugs were introduced, formulation of medicines was usually based on medicinal plants. Objective: To determine mycobiota and levels of Aspergillus section Flavi aflatoxins isolated from 50 samples of phytotherapeutic remedies currently commercialized in São Paulo, Brazil. Methods: Fifty (50) samples of phytotherapeutic remedies in the form of leaves (teas-25) and powders (capsules-25) were collected from August 2000 to July 2001. Filamentous fungi isolates were identified at the genera level in accordance with morphological characteristics and taxonomic criteria. Aflatoxins were performed by Thin-layer chromatography (TLC). Results: The microbiological analysis showed that 41 (82 percent) of phytotherapeutic remedies presented a fungal growth over 100 CFU/g. A total of 106 species of six different genera were isolated (Aspergillus, Penicillium, Mucor, Rhizopus and Alternaria). The genus Aspergillus was the predominant (60.5 percent) followed by Penicillium genus (20.0 percent). Aspergillus niger (30 percent) A. flavus (22 percent), A. fumigatus (6.5 percent) and A. parasiticus were the species of Aspergillus identified. It was observed that 13 (56.5 percent) of 23 A. flavus isolates and two A. parasiticus isolates produced aflatoxins. Conclusions: The contamination observed in most products and the high level of aflatoxigenic strains justify the concern regarding the execution of more careful analyzes of the commercialized phytotherapeutic remedies and the application of more rigorous laws that may warrant the quality of these products.

Micosociología: antecedentes históricos, evolución y proyecciones / Mycosociology: Historical background, evolution and projections

A lo largo de la historia numerosos investigadores han intentado estudiar la sociología de los hongos, también llamada micosociología que se refiere al estudio y la clasificación de las comunidades fúngicas, sus interrelaciones y su dependencia del medio. La mayoría de estos estudios han sido realizados en Europa, donde destacan los trabajos realizados por Darimont (1973) y García Bona (1977). Son pocos los estudios de carácter micosociológico que describen la interacción de ciertos macromicetos con especies de Nothofagus de América del Sur, destacando los de Godeas et al. (1993 a, b, c) en bosques de Nothofagus de Tierra del Fuego y el de Valenzuela et al. (1998). En esta revisión, se exponen los comienzos históricos de la micosociología, su evolución a lo largo de los años y cómo su enfoque ayuda a comprender el funcionamiento de las comunidades fúngicas. También, se destaca la importancia que tiene para nuestro país realizar estudios micosociológicos, especialmente, en bosques nativos dominados por Nothofagus.

Over time many researchers have tried to study the sociology of fungi, or mycosociology, meaning the study and classification of fungal communities, their inter-relations and their dependence on the medium. The majority of these studies have been carried out in Europe, notably the works of Darimont (1973) and García Bona (1977). There are few mycosociological studies which describe the interaction of certain macromycetes with South American Nothofagus species. Important studies are Godeas et al. (1993 a, b, c), in Nothofagus forests in Tierra del Fuego, and Valenzuela et al. (1998). This review examine the historical beginnings of mycosociology, its evolution over time and how its focus helps to understand the functioning of fungal communities. The autor also stress the importance for Chile of carrying out mycosociological studies, especially in native forests dominated by Nothofagus.

Estimación de la biomasa fúngica en un suelo del sudoeste de la provincia de Buenos Aires (Argentina) con una tinción directa con blanco de calcoflúor / Fungal biomass estimation in soils from southwestern Buenos Aires province (Argentina) using calcofluor white stain

Los microorganismos del suelo son vitales para el correcto funcionamiento de los ecosistemas, principalmente por su papel en el ciclado de nutrientes. La intensificación del uso del suelo y las prácticas agrícolas alteran negativamente la actividad microbiana. La biomasa fúngica es uno de los parámetros más utilizados para estudiar el impacto de las actividades agrícolas en la estructura y el funcionamiento del suelo. El objetivo del presente trabajo fue estimar la biomasa fúngica en un suelo del sudoeste bonaerense con el fin de obtener valores de referencia que permitan usar este parámetro como un indicador de cambios en el ecosistema y, por otro lado, demostrar que la metodología empleada es sensible a las variaciones en las condiciones climáticas. Se colectaron muestras de suelos durante 2 años consecutivos. Se prepararon frotis de suelo y se tiñeron con soluciones de distintas concentraciones de blanco de calcoflúor y luego se estimó la biomasa fúngica observando los frotis con microscopio de epifluorescencia. Los valores de biomasa fúngica estimados variaron entre 2,23 y 26,89 μg Cfúngico/g de suelo y estuvieron dentro del rango esperable para el tipo de suelo estudiado. La biomasa fúngica mostró una relación positiva con la temperatura y las precipitaciones. La metodología empleada resultó ser confiable, repetible y sensible a cambios en las condiciones climáticas. Los resultados podrían usarse como valores de referencia para estudiar la biomasa fúngica de suelos bajo distintas condiciones y emplearse como indicadores del impacto de las distintas prácticas agrícolas sobre el ecosistema.

Soil microorganisms are vital for ecosystem functioning because of the role they play in soil nutrient cycling. Agricultural practices and the intensification of land use have a negative effect on microbial activities and fungal biomass has been widely used as an indicator of soil health. The aim of this study was to analyze fungal biomass in soils from southwestern Buenos Aires province using direct fluorescent staining and to contribute to its use as an indicator of environmental changes in the ecosystem as well as to define its sensitivity to weather conditions. Soil samples were collected during two consecutive years. Soil smears were prepared and stained with two different concentrations of calcofluor, and the fungal biomass was estimated under an epifluorescence microscope. Soil fungal biomass varied between 2.23 and 26.89 μg fungal C/g soil, being these values in the range expected for the studied soil type. The fungal biomass was positively related to temperature and precipitations. The methodology used was reliable, standardized and sensitive to weather conditions. The results of this study contribute information to evaluate fungal biomass in different soil types and support its use as an indicator of soil health for analyzing the impact of different agricultural practices.

Aerobiología y biodeterioro del género Aspergillus Link en depósitos de tres instituciones patrimoniales cubanas / Aerobiology and biodeterioration of the genus Aspergillus Link in repositories of three Cuban heritage intitutions

Las especies del género Aspergillus Link constituyen importantes agentes biodeteriorantes de objetos de valor constructivo, cultural y patrimonial. En ambientes laborales, sus propágulos y metabolitos pueden provocar afecciones a la salud humana. Los estudios aeromicológicos pueden contribuir a minimizar el impacto de Aspergillus spp. en el biodeterioro y la calidad de vida. Se han caracterizado de acuerdo a su potencial biodeterió- geno y patogénico representantes de Aspergillus spp. en el aire de depósitos de materiales en tres instituciones patrimoniales cubanas. Las muestras se tomaron empleando un biocolector SAS súper 100. Se evaluó cualitativamente la actividad celulolítica, amilolítica y proteolítica de los aislados; así como la excreción de pigmentos y ácidos orgánicos. Se evaluó el crecimiento a 37°C y la secreción de hemolisinas. Aspergillus spp. constituyó parte considerable de la micobiota aérea viable en los depósitos con 22 especies correspondientes a 12 secciones, las de mayor variabilidad fueron Flavi y Restricti, mientras la primera y la sección Aspergillus fueron las mejor representadas. El 74 por ciento de las cepas mostraron alta capacidad para degradar los compuestos orgánicos evaluados y secretar ácidos, mientras el 34 por ciento excretó pigmentos de amarillo a pardo oscuro, se evidenció un amplio potencial biodeteriorante en general. Más del 30 por ciento de los aislados mostraron ser patógenos oportunistas peligrosos para el hombre al crecer a 37°C, secretar hemolisinas, y presentar un tamaño conidial que posibilita la depositación en alvéolos pulmonares. Las cepas con mayor impacto biodeteriorante y patogénico fueron las correspondientes a las Secciones Flavi, Nigri y la especie Aspergillus fumigatus Fresen.

Species of the genus Aspergillus Link cause damage to objects constructive, cultural and heritage value. In work environments, their propagules and metabolites may cause disorders to human health. Aerobiological studies can help minimize the impact of Aspergillus spp. in the biodeterioration and quality of life. Aspergillus spp. strains present in the air of the repositories in three Cuban heritage institutions were characterized according to their biodeteriogenic and pathogenic potential. Each isolated was classified taxonomically, cellulolytic, amylolytic and proteolytic activity as well as excretion of pigments and organic acids was evaluated. The capacity to grow at 37°C and secrete hemolysins was qualitatively evaluated. Aspergillus spp. was significant portion of viable air micobiota in the three repositories with 22 species corresponding to 12 sections, most variability were Flavi and Restricti, while the first section and Aspergillus were the best represented. The 74 percent of the strains showed high ability to degrade organic compounds evaluated and secrete acids, while 34 percent excreted pigments from yellow to dark brown, evidenced a broad biodeteriogenic potential in general. More than 30 percent of the isolates were shown to be dangerous opportunistic pathogens for humans to grow at 37°C, secrete hemolysin, and present a conidial size which enables the deposition pulmonary alveoli. Strains higher biodeteriogenic and pathogenic impact were those corresponding to the Flavi, Nigri sections and the species Aspergillus fumigatus Fresen.

Desinfectantes en aerosol como alternativa de control ambiental de los hongos anemófilos en viviendas / Disinfectant spray as an alternative environmental control of airborne fungi in houses

El objetivo de este trabajo fue evaluar el efecto de productos desinfectantes comerciales en aerosol sobre la carga de hongos ambientales en habitaciones de viviendas, teniendo en cuenta que las personas, principalmente los niños, pasan la mayor parte del día en las mismas. Además, se estudió, a través de encuestas, las preferencias del público a la hora de adquirir los desinfectantes comerciales ensayados. El número de UFC/placa fue variable dentro de la misma habitación para las distintas mediciones realizadas antes de la aplicación de los productos desinfectantes. Los porcentajes de inhibición del crecimiento fúngico fueron variables dependiendo del producto desinfectante ensayado y de la habitación muestreada. Dichos porcentajes no fueron diferentes significativamente entre los desinfectantes. Los valores de temperatura estuvieron dentro de un rango de 11 a 19,5 ºC y el porcentaje de humedad relativa varió entre un 64 a 85 por ciento. El 89 por ciento de los encuestados conocía la existencia de desinfectantes ambientales en aerosol. El 78 por ciento conocía el Lysoform®, seguido por Glade® antitabaco y Ayudin®. Dentro de los productos más utilizados, el 54 por ciento de los encuestados indicó el producto Lysoform®. La mayoría de los consumidores eligieron a los desinfectantes por lo que conocen y los medios de comunicación influyeron en el 29 por ciento de los consumidores al momento de elegir el desinfectante. Estos productos ensayados son recomendables en una habitación para disminuir la carga de hongos ambientales y así reducir el efecto perjudicial de estos hongos sobre las personas que la utilizan.

The aim of this study was to evaluate the effect of commercial disinfectant spray on the concentration of airborne fungi in bedrooms, considering that people, especially children, spend most of the day in them. Also, public preferences through surveys were studied when acquiring commercial disinfectants tested. The number of CFU/Petri dish was variable within the same bedroom for the different measurements performed before application of the disinfectant. Percent inhibition of fungal growth were variable, depending on the disinfecting product tested and sampled bedroom. These percentages were not significantly different between the disinfectants. The temperature values were within a range of 11 to 19.5° C and relative humidity percentage ranged from 64 to 85 percent. Eighty nine percent of respondents knew about environmental disinfectant spray. Seventy-eight percent knew about Lysoform®, followed by antismoking Glade® and Ayudin®. Among the most widely used products, 54 percent of respondents said the Lysoform® product was the most used. Most consumers chose known disinfectants and, when choosing a disinfectant, the media influenced 29 percent of consumers. The used products are recommended to be used in a bedroom to decrease the number of airborne fungi and thus reduce the harmful effect of these fungi on the people that use this room.

Micobiota parasitária de escleródios de Sclerotinia sclerotiorum isolada de solos da fronteira oeste do Rio Grande do Sul / Parasitic mycoflora of sclerotia of Sclerotinia sclerotiorum isolated from soils of the western frontier of the state of Rio Grande do Sul, Brazil

O objetivo do presente trabalho foi identificar a diversidade da micobiota parasitária e o tempo decorrido até o início do parasitismo de escleródios de Sclerotinia sclerotiorum em solos da fronteira oeste do Rio Grande do Sul (RS). Foram coletados solos agrícolas e não agrícolas de cinco localidades na fronteira oeste do Rio Grande do Sul. O isolamento dos fungos foi realizado por meio do teste de iscas, sendo que as iscas foram os escleródios do fitopatógeno, os quais permaneceram enterrados no solo durante 15, 30 e 60 dias. Os fungos foram identificados em nível de gênero. Os gêneros fúngicos encontrados parasitando escleródios foram: Trichoderma, Penicillium, Aspergillus, Fusarium, Pythium (Cromista), Clonostachys rosea (Gliocladium), Chaetomium, Curvularia, Phytophthora (Cromista), Rhizopus, Lichtheimia (Absidia), Mucor, Acremonium, Periconia e Isaria.Observou-se que os escleródios foram parasitados pelos fungos até 15 dias depois de serem enterrados, e deste período em diante apenas a frequência de ocorrência dos fungos mudou. Concluiu-se que os gêneros fúngicos Isaria, Periconia, Acremonium, Lichtheimia (Absidia), Phytophthora, Curvularia e Chaetomium são encontrados em solos agrícolas e não agrícolas na fronteira oeste do RS, e não foram citados anteriormente em outros trabalhos. Existe diversidade de fungos parasitários de escleródios de S. sclerotiorum nos solos da fronteira oeste do RS, os quais podem ser utilizados em experimentos para compor um programa de controle biológico desse fitopatógeno. Os escleródios são parasitados pelos fungos durante até 15 dias, após os mesmos serem enterrados no solo.(AU)

The aim of this study was to identify the diversity of the parasitic mycoflora and to identify the necessary time until the beginning of the parasitism of sclerotia of Sclerotinia sclerotiorum in soils of the western frontier of the state of Rio Grande do Sul (RS), Brazil. Agricultural and non agricultural soils of five locations in the western frontier of Rio Grande do Sul were collected. The fungi isolation was performed by testing baits, and baits were the sclerotia of the phytopathogen, which remained buried in the soil for 15, 30 and 60 days. The fungi were identified at the genus level. The following fungal genera were found parasitizing sclerotia: Trichoderma, Penicillium, Aspergillus, Fusarium, Pythium (Cromista) Clonostachys rosea (Gliocladium), Chaetomium, Curvularia, Phytophthora (Cromista), Rhizopus, Lichtheimia (Absidia), Mucor, Acremonium, Periconia and Isaria. It was observed that the sclerotia were parasitized by fungi until 15 days after they had been buried. And after this period, only the frequency of occurrence of fungi has changed. It was concluded that the fungal genera Isaria, Periconia, Acremonium, Lichtheimia (Absidia), Phytophthora, Curvularia and Chaetomium are found both in agricultural and non-agricultural soils on the western frontier of RS, but they were not previously cited in other works. There is diversity of parasitic fungi of sclerotia of S. sclerotiorum in the soils of the western frontier of the RS, which can be used in experiments to compose biological control programs for this pathogen. The sclerotia are parasitized by fungi until 15 days after being buried underground.(AU)