Dos interesantes contaminantes fúngicos filamentosos en muestras clínicas positivas a Trychophyton rubrum: una situación de curiosidad morfológica / Two interesting filamentous fungal contaminants in clinical samples positive to Trychophyton rubrum: a situation of morphological curiosity

Las dermatofitosis corresponden a un grupo de enfermedades micóticas comunes en piel y fanéreas, donde Trichophyton rubrum es el agente causante más frecuente a nivel mundial y presente en nuestros 2 casos de pacientes masculinos con estas micosis, una en uñas y la otra en piel. Sin embargo, el enfoque de esta publicación se basa principalmente en la presencia de 2 interesantes contaminantes (uno en cada caso clínico) presentes solo en los cultivos de las primeras siembras como saprófitos y por ende como propágulos de dispersión, asociados al ambiente y sin intervención clínica demostrada en ambas micosis. La descripción morfofisiológica de estos 2 contaminantes Metarhizium purpureo-genum(similis) y Monascus ruber fue más bien una curiosidad esencial que el micólogo clínico adquiere en su contínua formación y ante la posibilidad de infecciones mixtas, pudiendo conjugar sus hallazgos junto al análisis taxonómico y los factores geográficos y edáficos asociados a su distribución. (AU)

Dermatophytoses belongs to a group of common mycotic diseases in skin and pharynals, where Trichophyton rubrum is the most frequent causative agent worldwide and present in our 2 cases of male patients with these mycoses, one in nails and the other in skin. However, the focus of this publication is mainly about the presence of 2 interesting contaminants (one in each clinical case) present only in the crops of the first sowings as saprophytes and therefore as dispersal propagules, associated with the environment and without clinical intervention demonstrated in both mycoses. The morphophysiological description of these 2 contaminants, Metarhizium purpureogenum (similis) and Monascus ruber was rather an essential curiosity that the clinical mycologist acquires in his continuous training and in the face of the possibility of mixed infections, being able to combine his findings together with the taxonomic analysis and the geographic and edaphic factors associated with its distribution. (AU)

Effects of nutrient additives and incubation period on sporulation and viability of the entomopathogenic fungus, Metarhizium anisopliae (Hypocreales: Clavicipitaceae)

Aims@#Metarhizium anisopliae is an entomopathogenic fungus (EPF) that exists naturally in the environment and potentially be used as a biological control agent against many insect pests. This study aims to evaluate the effect of nutrient additives on the yield and viability of M. anisopliae spore and to determine the optimum incubation period for maximum spore production.@*Methodology and results@# In this study, M. anisopliae was cultivated by solid-state fermentation using rice as a growth medium. Three different nutrient additives were examined which aimed to maximize the production of M. anisopliae spores. Among the three nutrient additives evaluated, yeast (1.84 ± 0.04 g) supported better growth and spore production than molasses (0.58 ± 0.04 g) and palm oil (0.47 ± 0.09 g). The incubation period between 2-6 weeks produced higher spore yield (0.97 ± 0.02 g spores) at week 4 with a better spore viability (86.30 ± 0.45%) at week 2. @*Conclusion, significance and impact of study@#Hence, it is suggested that the optimum incubation period is between 2 and 6 weeks after inoculation, and M. anisopliae could be mass produced in large quantities on rice substrate with the addition of yeast as the nutrient additives.

Efecto de la combinación de Metarhizium anisopliae y Gliocladium virens sobre la oviposición, eclosión y emergencia de Aedes aegypti / Effect of the combination of Metarhizium anisopliae and Gliocladium virens on the oviposition, hatching and emergency of Aedes aegypti

Resumen: Objetivo: Evaluar el efecto de la combinación de Metarhizium anisopliae y Gliocladium virens, ambos con Aqua Reslin Super, sobre oviposición, eclosión y emergencia de Aedes aegypti. Material y métodos: Se realizaron evaluaciones para determinar el efecto de los tratamientos impregnados en papel filtro y expuestos dentro de recipientes de plástico sobre la oviposición, eclosión y emergencia de Aedes aegypti. Resultados: Los resultados indicaron que las combinaciones hongo e insecticida no afectaron el comportamiento de oviposición, pero sí la eclosión de los huevos y la emergencia del adulto. Conclusión: Con los resultados se puede concluir que la combinación de hongos + insecticida puede ser una buena opción para aplicarse en sitios de oviposición con miras al desarrollo de una ovitrampa letal.

Abstract: Objective: To evaluate the effect of the combination of Metarhizium anisopliae and Gliocladium virens, both with Aqua Reslin Super, on the oviposition, hatching and emergence of Aedes aegypti. Materials and methods: Evaluations were carried out to determine the effect of treatments impregnated on filter paper and exposed within plastic containers on the oviposition, hatching and emergency of Aedes aegypti. Results: The results indicated that the fungus and insecticide combinations did not affect the oviposition behavior, but if the hatching of the eggs and the adult's emergency. Conclusion: With the results it can be concluded that the combination of fungi + insecticide can be a good option to be applied in oviposition sites with a view to the development of a lethal ovitrap.

Encapsulation of Metarhizium anisopliae spores with zeolite nanoparticles and magnesium silicate nanoparticles against mortality and lethal times of Crocidolomia pavonana larvae

Aims@#Cruciferae crops failure such as cabbage generally due to heavy attacks by of Crocidolomia pavonana larvae. Metarhizium anisopliae is a species of fungus that can infect more than 200 insect pests including C. pavonana larvae. However, the direct application of M. anisopliae spore in the field is susceptible to UV rays from the sun, which can decrease the efficiency of M. anisopliae. Thus, this study aims to investigate the encapsulation performance of M. anisopliae spore with zeolite and magnesium silicate nanoparticles in terms of mortality and lethal time against 3 rd instar C. pavonana larvae.@*Methodology and results@#Zeolite and magnesium silicate nanoparticles were chosen because they are non-toxic, environmentally friendly and able to maintain moisture, so that they are best used as protective materials for spores. The experiment was designed by Randomized Block Design (RBD) with a single factor. The obtained results showed that encapsulation of M. anisopliae spores with zeolite nanoparticles coating agent increased the mortality rate to 92.5% and accelerated the lethal time up to 1.075 days compared to only spore correspondingly 27.5% and 2.235 days. The M. anisopliae spores encapsulated with magnesium silicate nanoparticles also increased the mortality to 85.0% and accelerated the larval lethal time up to 1.150 days.@*Conclusion, significance and impact of study@#Metarhizium anisopliae spores that encapsulated with zeolite nanoparticles coating received higher mortality and faster lethal time to C. pavonana compared to those encapsulated with magnesium silicate nanoparticles. The encapsulation formulation of these two coatings can be used as bioinsecticide in controlling C. pavonana larvae.

In vitro efficacy of two commercial products of Metarhizium anisopliae s.l. for controlling the cattle tick Rhipicephalus microplus / Eficácia in vitro de dois produtos comerciais de Metarhizium anisopliae s.l. no controle do carrapato bovino Rhipicephalus microplus

Abstract The effects of two different products - Metarril® SP Organic (dry conidia) and Metarril® SC Organic (emulsifiable concentrated conidia in vegetable oil) - on eggs, larvae and Rhipicephalus microplus engorged females were here explored. Three concentrations (108, 107, and 106 conidia mL-1) for both products were prepared in water + 0.1% Tween® 80 (v/v); afterward, bioassays were carried out for all R. microplus stages by immersion in suspensions (Metarril® SP) or formulations (Metarril® SC). Metarril® SP suspensions showed low efficacy and did not affect biological parameters of treated engorged females; for eggs and larvae, only slight decreases in hatchability and larvae population were observed. Despite a delay in germination, Metarril® SC presented better results; for females, reductions in Egg Mass Weight (EMW) and Egg Production Index (EPI) were reported. On eggs, 108 conidia mL-1 increased Incubation Period (IP), shortened Hatching Period (HP) and decreased hatchability by up to 61%; for larvae, 107 and 108 conidia mL-1 reached 99.6 and 100% larval mortality respectively, 10 days after fungal exposure. Thus, further studies involving the use of oil-based formulations for ticks such as Metarril® SC need to be performed, especially to control the most susceptible stages (eggs and larvae).

Resumo No presente trabalho, os efeitos de dois diferentes produtos foram avaliados - Metarril® SP Organic (conídios secos) e Metarril® SC Organic (conídios concentrados em óleo vegetal) - para ovos, larvas e fêmeas ingurgitadas de Rhipicephalus microplus. Três concentrações (108, 107 e 106 conídios mL-1) para cada produto foram preparadas em água + Tween® 80 0,1% (v/v); os bioensaios foram realizados para todos os estágios de R. microplus por imersão nas suspensões (Metarril® SP) ou formulações (Metarril® SC). Metarril® SP não afetou os parâmetros biológicos das fêmeas, demonstrando assim baixa eficácia; para ovos e larvas, foram observadas discretas diminuições na eclodibilidade e na população de larvas. Apesar de um atraso na germinação, Metarril® SC apresentou melhores resultados; para as fêmeas, foram detectadas reduções no Peso da Massa de Ovos (PMO) e no Índice de Produção de Ovos (IPO). Para os ovos, a concentração de 108 conídios mL-1 aumentou o Período de Incubação (PI), reduziu o Período de Eclosão (PE) e também o da eclodibilidade em até 61%; para larvas, 107 e 108 conídios mL-1 atingiram 99,6 e 100% de mortalidade larval, respectivamente, 10 dias após a exposição fúngica. Com isso, estudos adicionais que envolvem o uso de formulações à base de óleo para carrapatos, como Metarril® SC, precisam ser realizados, especialmente para controlar os estágios mais suscetíveis (ovos e larvas).

Control of Haematobia irritans and Stomoxys calcitrans with Metarhizium anisopliae in naturally infested cattle / Control de Haematobia irritans y Stomoxys calcitrans con Metarhizium anisopliae en ganado naturalmente infestado

ABSTRACT Objective. Assess the efficacy of Metarhizium anisopliae strain 135 (Ma135) applied by aspersion to reduce simultaneous infestation of Haematobia irritans and Stomoxys calcitrans in naturally infested cattle maintained in a production system which combine grazing and confinement in pens. Materials and methods. Was applied an aqueous formulation of Ma135 (1x108 conidia/ml) on a nine cows group in six occasions with seven days interval, while the control group received the same formulation without conidial content. The infestation index was estimated daily for each fly independently; the effectiveness of the formulation was calculated using the Abbott's formula. Results. The Ma135 formulation had a reduction in the fly population of 58% for H. irritans and 69% for S. calcitrans, taking into consideration the six study weeks, in addition to causing no negative effects on animal health. An Infestation index reduction was observed from the first week post-treatment (p<0.05) and maintained this trend throughout the study. Conclusions. The present study has demonstrated the potential of strain Ma135 to reduce the simultaneous infestation of both hematophagous flies in cattle under conditions of natural infestation.

RESUMEN Objetivo. Evaluar la eficacia de la cepa 135 de Metarhizium anisopliae (Ma135) aplicada por aspersión para reducir la infestación simultánea de Haematobia irritans y Stomoxys calcitrans en ganado naturalmente infestado mantenido en un sistema de producción de leche que combina el pastoreo y el confinamiento en corrales. Materiales y métodos. Se aplicó una formulación acuosa de Ma135 (1x108 conidios/ml) a un grupo de nueve vacas en seis ocasiones con un intervalo de siete días, mientras que el grupo control recibió una solución compuesta por agua, Monooleato de polioxietileno sorbitán en solución 0.01% y un adyuvante agrícola al 0.1%. Se estimó diariamente el índice de infestación para cada mosca de forma independiente; la efectividad de la formulación se calculó usando la fórmula de Abbott. Resultados. La formulación de Ma135 tuvo una eficacia en el control de la infestación de 58% para el caso de H. irritans y de 69% para S. calcitrans, tomando en cuenta las seis semanas de estudio, además de que no causó ningún efecto negativo en la salud de los animales. La reducción del índice de infestación se observó desde la primera semana post-tratamiento (p<0.05) y mantuvo esta tendencia durante todo el estudio. Conclusiones. El presente estudio ha demostrado el potencial de la cepa Ma135 para reducir la infestación simultánea de ambas moscas hematófagas en el ganado bajo condiciones de infestación natural.

Pathogenicity of Metarhizium anisopliae and Beauveria bassiana fungi to Tetranychus ludeni (Acari: Tetranychidae) / Patogenicidade de fungos Metarhizium anisopliae e Beauveria bassiana ao Tetranychus ludeni (Acari: Tetranychidae)

The use of acaricides is the main control method for Tetranychus ludeni (Acari: Tetranychidae) in horticultural crops. This mite has been recorded causing damage to sweet potato (Ipomoea batatas L.). The use of pathogenic fungi is an alternative to chemical control. The objective was to evaluate the pathogenicity of the fungi Metarhizium anisopliae (Meetch) Sorok, and Beauveria bassiana (Bals.) Vuill. to phytophagous mite T. ludeni in sweet potato plants. Conidial suspensions of M. anisopliae and B. bassiana, at concentrations of 106 and 107 con.mL-1 were applied to sweet potato leaves. After 24 hours, five females of T. ludeni newly emerged were released on the leaves. The bioassay was arranged in a completely randomized design with factorial arrangement 2 × 2 (two species of fungi and two concentrations) plus the control (distilled water), with 10 repetitions per treatment. The evaluation consisted of observing of three biological parameters of the mite: mortality, oviposition, and repellency, after 24, 48, 72, and 96 hours of contact with the fungi. The isolates of M. anisopliae cause high mortality rates of T. ludeni in laboratory. Beauveria bassiana has the potential to suppress future generations of mite, reducing its oviposition rate. Repellency behavior was not observed.(AU)

O uso de acaricidas é o principal método de controle de Tetranychus ludeni (Acari: Tetranychidae) em cultivos hortícolas. Esse ácaro foi registrado causando danos em batata-doce (Ipomoea batatas (L.). A utilização de fungos patogênicos é uma alternativa ao controle químico. O objetivo foi avaliar a patogenicidade dos fungos Metarhizium anisopliae (Meetch) Sorok. e Beauveria bassiana (Bals.) Vuill. ao ácaro fitófago T. ludeni em batata-doce. Suspensões conidiais de M. anisopliae e B. bassiana, nas concentrações de 106 e 107 con.mL-1, foram aplicadas sobre folhas de batata-doce. Após 24 horas, cinco fêmeas recém-emergidas foram liberadas sobre as folhas. O bioensaio foi inteiramente randomizado, com arranjo fatorial 2 × 2 (duas espécies de fungos e duas concentrações) e controle (água destilada), com 10 replicações por tratamento. A avaliação consistiu na observação de três parâmetros biológicos do ácaro: mortalidade, oviposição e repelência, após 24, 48, 72 e 96 horas de contato com os fungos. Os isolados de M. anisopliae causam altas taxas de mortalidade de T. ludeni em laboratório. Beauveria bassiana tem potencial para suprimir futuras gerações do ácaro, reduzindo a taxa de oviposição. Comportamento de repelência nos ácaros não foi observado.(AU)

Compatibility of chemical insecticides and entomopathogenic fungi for control of soybean defoliating pest, Rachiplusia nu / Compatibilidad de insecticidas químicos y hongos entomopatógenos para el control de Rachiplusia nu, plaga defoliadora de la soja

Rachiplusia nu (Guenée) (Lepidoptera: Noctuidae) is one of the major lepidopteran pests defoliating soybeans (Glycine max Merrill) in Argentina. The combined use of chemical insecticides and entomopathogenic fungi is a promising pest-control option to minimize adverse chemical effects. In this work, we evaluated the interactions between five insecticides-two being considered biorational-and five fungal entomopathogenic strains under laboratory conditions in order to determine the possible usefulness of combinations of these agents against R. nu. The insecticides were tested for compatibility at four doses by in vitro bioassay and for the lethality of R. nu by inoculations at three doses. Fungal strains were applied at 1 x 10(8), 1 x 10(6), and 1 x 10(4) conidia/ml. The combinations of those insecticides with Beauveria bassiana (LPSc 1067, LPSc 1082, LPSc 1098), Metarhizium anisopliae (LPSc 907), and Metarhizium robertsii (LPSc 963) caused higher R. nu-larval mortalities than any of the individual agents alone. We observed significant differences in the in vitro conidial viability, vegetative growth, and conidia production of the five strains of entomopathogenic fungi exposed to different doses of the chemical insecticides. The combination gamma-cyhalothrin-LPSc-1067 caused the highest percent mortality of R. nu larvae, with synergism occurring between the two agents at 50% and 25% of the maximum field doses.

Rachiplusia nu (Guenée) (Lepidoptera: Noctuidae) es una de las principales plagas de lepidópteros defoliadores del cultivo de soja (Glycine max Merrill) en Argentina. El uso combinado de insecticidas químicos y hongos entomopatógenos es una opción de control de plagas prometedora para minimizar los efectos químicos adversos. En este trabajo se evaluaron las interacciones entre 5 insecticidas -2 de ellos considerados biorracionales-- y 5 cepas fúngicas entomopatógenas en condiciones de laboratorio, para determinar la posible utilidad de combinaciones de estos agentes frente a R. nu. Se evaluó la compatibilidad de los insecticidas a 4 dosis mediante bioensayos in vitro y la letalidad de aquellos sobre R. nu mediante inoculaciones a 3 dosis. Las cepas fúngicas se evaluaron a concentraciones de 1 x 10(8), 1 x 10(6) y 1 x 10(4) conidios/ml. Las combinaciones de estos insecticidas con Beauveria bassiana (LPSc1067, LPSc 1082, LPSc 1098), Metarhizium anisopliae (LPSc 907) y Metarhizium robertsii (LPSc 963) causaron una mayor mortalidad de larvas de R. nu que cualquiera de los agentes individuales. Asimismo, se observaron diferencias significativas en la viabilidad de los conidios in vitro, el crecimiento vegetativo y la producción de conidios de las 5 cepas de hongos entomopatógenos expuestos a diferentes dosis de los insecticidas químicos. La combinación gamma-cialotrina-LPSc-1067 causó el mayor porcentaje de mortalidad de larvas de R. nu, con un efecto de sinergismo entre los 2 agentes al 50 y el 25% de las dosis de campo recomendadas por el fabricante.

Compatibility of Metarhizium anisopliae with liposoluble photoprotectants and protective effect evaluation against solar radiation / Compatibilidade de Metarhizium anisopliae com fotoprotetores lipossolúveis e avaliação do efeito protetor contra a radiação solar

The aim of this study was to analyze the compatibility of the Metarhizium anisopliae JAB 68 isolate with liposoluble photoprotectants and the photoprotection conferred to its conidia. The fungus was cultivated in a medium with the photoprotectants Neo Heliopan® BB, Neo Heliopan® E 1000, Eusolex® 6007 and Neo Heliopan® AV in various concentrations. Vegetative growth, sporulation and germination were assessed to determine compatibility. Then, conidia were suspended in oil containing the compatible photoprotectants, in their respective concentrations, and exposed to 0-7 hours of light from a solar simulator operating at 1,000 W m-2. Neo Heliopan BB® and Neo Heliopan® E 1000 were compatible with M. anisopliae at 2% and 1%, respectively. Eusolex® 6007 and Neo Heliopan® AV were compatible up to 8% a 7%, respectively. Conidia exposed to radiation from the solar simulator, with photoprotectant-free oil as its carrier, presented germination of 78.12% after five hours. When adding any photoprotectant to the conidia oily suspension, the germination percentage was always higher than that obtained in the control. Neo Heliopan® BB at 1 and 2% and Neo Heliopan® AV at 0.5% increased the germination. After 7 hours of exposure the germination percentages were 93.77, 94.55 and 98.82%, respectively. Eusolex® 6007 was not as efficient as previous products, but in the same exposure period was able to protect the conidia (88.60% germination) when used at a 1% concentration. Adding photoprotectants in oily formulations of M. anisopliae protects against the harmful effects of UV radiation contributing to the fungus efficiency in the field.

O objetivo desse trabalho foi analisar a compatibilidade do isolado JAB 68 de Metarhizium anisopliae com fotoprotetores lipossolúveis e a fotoproteção conferida aos seus conídios. O fungo foi cultivado em meio contendo os fotoprotetores Neo Heliopan® BB, Neo Heliopan® E 1000, Eusolex® 6007, Neo Heliopan® AV em várias concentrações. O crescimento vegetativo, a esporulação e a germinação foram avaliados para determinar a compatibilidade. Conídios foram suspensos em óleo contendo os fotoprotetores compatíveis, em suas respectivas concentrações, e submetidos por 0 a 7 horas à luz de um simulador solar na potência de 1.000W m-2. Neo Heliopan BB® e Neo Heliopan® E 1000 foram compatíveis com M. anisopliae à 2% e 1%, respectivamente. Eusolex® 6007 e Neo Heliopan® AV foram compatíveis até 8% e 7%, respectivamente. Conídios de M. anisopliae expostos à radiação do simulador solar, tendo óleo isento de fotoprotetor como veículo, apresentaram germinação de 78,12% após cinco horas. Adicionando-se qualquer fotoprotetor à suspensão oleosa de conídios a porcentagem de germinação foi sempre maior que a obtida no controle. Neo Heliopan® BB a 1 e 2% e Neo Heliopan® AV a 0,5% incrementaram a germinação. Após 7 horas de exposição as porcentagens de germinação foram de 93,77, 94,55 e 98,82%, respectivamente. Eusolex® 6007 não foi tão eficiente quanto os produtos anteriores, mas no mesmo período de exposição foi capaz de proteger os conídios (88,60% de germinação) quando usado na concentração de 1%. A adição de fotoprotetores em formulações oleosas de M. anisopliae confere proteção aos raios UV, contribuindo com a eficiência do fungo no campo.

Agrobacterium-Mediated Co-transformation of Multiple Genes in Metarhizium robertsii

Fungi of the Metarhizium genus are a very versatile model for understanding pathogenicity in insects and their symbiotic relationship with plants. To establish a co-transformation system for the transformation of multiple M. robertsii genes using Agrobacterium tumefaciens, we evaluated whether the antibiotic nourseothricin has the same marker selection efficiency as phosphinothricin using separate vectors. Subsequently, in the two vectors containing the nourseothricin and phosphinothricin resistance cassettes were inserted eGFP and mCherry expression cassettes, respectively. These new vectors were then introduced independently into A. tumefaciens and used to transform M. robertsii either in independent events or in one single co-transformation event using an equimolar mixture of A. tumefaciens cultures. The number of transformants obtained by co-transformation was similar to that obtained by the individual transformation events. This method provides an additional strategy for the simultaneous insertion of multiple genes into M. robertsii.

Entomopathogenic Fungi as Dual Control Agents against Both the Pest Myzus persicae and Phytopathogen Botrytis cinerea

The green peach aphid (Myzus persicae), a plant pest, and gray mold disease, caused by Botrytis cinerea, affect vegetables and fruit crops all over the world. To control this aphid and mold, farmers typically rely on the use of chemical insecticides or fungicides. However, intensive use of these chemicals over many years has led to the development of resistance. To overcome this problem, there is a need to develop alternative control methods to suppress populations of this plant pest and pathogen. Recently, potential roles have been demonstrated for entomopathogenic fungi in endophytism, phytopathogen antagonism, plant growth promotion, and rhizosphere colonization. Here, the antifungal activities of selected fungi with high virulence against green peach aphids were tested to explore their potential for the dual control of B. cinerea and M. persicae. Antifungal activities against B. cinerea were evaluated by dual culture assays using both aerial conidia and cultural filtrates of entomopathogenic fungi. Two fungal isolates, Beauveria bassiana SD15 and Metarhizium anisopliae SD3, were identified as having both virulence against aphids and antifungal activity. The virulence of these isolates against aphids was further tested using cultural filtrates, blastospores, and aerial conidia. The most virulence was observed in the simultaneous treatment with blastospores and cultural filtrate. These results suggest that the two fungal isolates selected in this study could be used effectively for the dual control of green peach aphids and gray mold for crop protection.

Sensibilidade de isolados de fungos entomopatogênicos às radiações solar, ultravioleta e à temperatura / Sensibility of isolates of entomopathogenic fungi to solar radiation, ultraviolet rays and temperature

Este trabalho teve por objetivo avaliar a sensibilidade de isolados dos fungos Metarhizium anisopliae (Metsch.) Sorok. e Beauveria bassiana (Bals). Vuill. ao efeito das radiações solar e ultravioleta e da temperatura. Conídios dos isolados foram expostos, por vários períodos, aos raios de um simulador solar em diversas irradiâncias e a uma lâmpada de raios ultravioleta germicida. Os conídios do isolado de M. anisopliae foram também expostos às temperaturas de 19,5; 24,2 e 31,0ºC, e os do isolado de B. bassiana a 19,4; 20,8 e 28,3ºC, e 18,7; 23,8 e 30,9ºC. Avaliou-se a germinação de conídios pelo teste de viabilidade. Os isolados dos fungos se mostraram bastantes sensíveis aos raios do simulador solar e aos raios ultravioleta. A germinação de ambos sofreu significativa redução a partir de 30 minutos de exposição à radiação do simulador solar. O efeito mais severo foi evidenciado pelo isolado de B. bassiana, com grande redução da germinação dos conídios em todas as irradiâncias testadas. A sensibilidade à radiação ultravioleta também foi grande, pois ocorreu acentuada redução da germinação dos conídios do isolado de M. anisopliae (38,2%) e de B. bassiana (65%) já aos 30 segundos de exposição. A temperatura afetou a viabilidade de ambos os fungos. Temperaturas entre 23,8 e 31ºC favoreceram a germinação dos conídios, enquanto temperaturas próximas de 20ºC dificultaram a germinação.(AU)

This study aimed to access the sensibility of isolates of the fungus Metarhizium anisopliae (Metsch.) Sorok. and Beauveria bassiana (Bals.) Vuill. to the effect of solar and ultraviolet radiation and temperature. Conidia were exposed for various periods to the rays from a solar simulator at various irradiances, and to light germicidal ultraviolet rays. Conidia of the isolate of M. anisopliae were also exposed to temperatures of 19.5, 24.2 and 31.0ºC and the isolate of B. bassiana to 19.4, 20.8 and 28.3ºC, and also to 18.7, 23.8 and 30.9ºC. The germination of conidia was evaluated by the viability test. The fungal isolates showed to be very sensitive to the solar simulator and ultraviolet rays. Germination of both was significantly decreased starting from 30 minutes of exposure to the rays of the solar simulator. The most severe effect was evidenced by the isolate of B. bassiana with great reduction in conidia germination in all the tested irradiances. Sensitivity to ultraviolet radiation was also great, showing a marked reduction in the germination of M. anisopliae (38.2%) and B. bassiana (65%) conidia after 30 seconds of exposure. The temperature affected the viability of both fungi. Temperatures ranging of 23.8 to 31ºC favor the germination of conidia while temperatures around 20ºC constrained germination.(AU)

Ocurrencia natural de hongos entomopatógenos en suelos cultivados con yerba mate (Ilex paraguariensis St. Hil.) en Misiones, Argentina / Natural ocurrence of entomopathogenic fungi in soils cultivated with Paraguay tea (Ilex paraguariensis St. Hil.) in Misiones, Argentina

Este trabajo tuvo por objetivo aislar, identificar y caracterizar morfológicamente hongos entomopatógenos presentes en suelos cultivados con yerba mate (Ilex paraguariensis St. Hil.). Se realizó una prospección de hongos entomopatógenos nativos a partir de 40 muestras de suelos cultivados con yerba mate ubicados en la provincia de Misiones, República Argentina, desde mayo de 2008 hasta junio de 2010. Las muestras de suelo se obtuvieron en cercanías de plantas de yerba mate, se aplicó la técnica de dilución seriada en medios de cultivos selectivos. Después de obtenidos los cultivos puros, los hongos fueron caracterizados macroscópica y microscópicamente, y se los identificó mediante el uso de claves taxonómicas específicas. Se aislaron 29 cepas fúngicas pertenecientes a las especies Beauveria bassiana (n = 17), Metarhizium anisopliae (n = 2) y Purpureocillium lilacinum (n = 10)

This study aimed to morphologically isolate, identify and characterize entomopathogenic fungi present in soils cultivated with Paraguay tea (Ilex paraguariensis). A survey of native entomopathogenic fungi was conducted from 40 soil samples grown with Paraguay tea in the province of Misiones, Argentina, from May 2008 to June 2010. The soil dilution plate methodology on selective culture media was used to isolate microorganisms. Taxonomic identification was performed using macroscopic and microscopic characters and specific keys. Twenty nine strains, belonging to the species Beauveria bassiana (n = 17), Metarhizium anisopliae (n = 2) and Purpureocillium lilacinum (n = 10) were isolated and identified

In vitro assay on homeopathic solutions on Metarhizium anisopliae (Metsch) Sorok (Ascomycota: Clavicipitaceae) / Efeito in vitro de soluções homeopáticas sobre Metarhizium anisopliae (Metsch) Sorok (Ascomycota: Clavicipitaceae)

This study aimed to evaluate the in vitro effect of homeopathic solutions Arsenicum Album 24CH; Calcarea carbonica 30CH; Kali iodatum 100CH;Phosphorus 3CH; Silicea 30CH; Staphysagria 6, 30 and 100CH; Spodoptera frugiperda 30CH; Sulphur 100 and 200CH and Thuya occidentalis 200CH on biological parameters of the fungus Metarhizium anisopliae . The solutions were diluted in sterile distilled water (0.1%) and were sprayed on the previously inoculated fungus on PDA culture medium. Germination, colony forming units, vegetative growth, conidial production and insecticidal activity of the fungus against larvae of Diatraea saccharalis (Lepidoptera: Crambidae) were evaluated. Homeopathic solutions did not affect negatively the parameters evaluated. Thus, all treatments were considered compatible to the fungus M. anisopliae.(AU)

O presente trabalho teve por objetivo avaliar o efeito in vitro das soluções homeopáticas Arsenicum album 24CH;Calcarea carbonica 30CH; Kali iodatum100CH; Phosphorus 3CH; Silicea 30CH; Staphysagria 6, 30 e 100CH; Spodoptera frugiperda 30CH; Sulphur 100 e 200CH eThuya occidentalis 200CH sobre parâmetros biológicos do fungo Metarhizium anisopliae. As soluções medicamentosas foram diluídas em água destilada (esterilizada) (0,1%) e pulverizadas sobre o fungo previamente inoculado no meio de cultura BDA. Os parâmetros biológicos avaliados foram: germinação de conídios, unidades formadoras de colônias, crescimento vegetativo, produção de conídios e atividade inseticida contra lagartas de Diatraea saccharalis (Lepidoptera: Crambidae). As soluções não afetaram os parâmetros avaliados. Todos os tratamentos foram considerados compatíveis ao M. anisopliae .(AU)

Confirmation of Two Undescribed Fungal Species from Dokdo of Korea Based on Current Classification System Using Multi Loci

Using dilution plating method, 47 fungal isolates were obtained from a soil sample collected from Dokdo in the East Sea of Korea in 2013. In this study, two fungal isolates, EML-MFS30-1 and EML-DDSF4, were confirmed as undescribed species, Metarhizium guizhouense and Mortierella oligospora in Korea based on current classification system using multi loci including rDNA internal transcribed spacer, large subunit, small subunit, and beta-tubulin (BTUB) genes. Herein, detailed morphological descriptions on characters of the undescribed fungal species as well as their molecular phylogenetic status are provided with comparisons to related species.

Liquid production of entomopathogenic fungi and ultraviolet radiation and temperature effects on produced propagules / Produção líquida de fungos entomopatogênicos e efeitos da radiação ultravioleta e da temperatura sobre os propágulos produzidos

The purpose of this paper was to evaluate the liquid culture media for the production of Metarhizium anisopliae (IBCB 425) and Beauveria bassiana (IBCB 66), as well as the tolerance of these seedlings to the ultraviolet action and to the temperature. Twelve treatments composed of combinations between carbon and nitrogen concentrations were assessed. In order to determine the effect of ultraviolet radiation, plates with blastos-pores were exposed to it for 25 and 50 seconds. To determine the temperature efect, blastospores from culture media were exposed to 20, 25, 30 and 35°. For the virulence experiments, caterpillars of Diatraea saccharalis were sprayed with 2 mL of fungal suspension with the aid of a Potter tower. Te best media for M. anisopliae are 16.00 g (carbon) + 7.00 g (nitrogen) and 14.40 g (carbon) + 7.00 g (nitrogen), whereas for B. bassiana: 20.00 g (carbon) + 6.30 g (nitrogen) and 20.00 g (carbon) + 7.00 g (nitrogen). Te longer the exposure to ultraviolet radiation, the smaller the number of colonies. At 35°, there is a significant decrease in the formation of colonies. Te produced seedlings of fungi are pathogenic to D. saccharalis.(AU)

O objetivo deste trabalho foi avaliar os meios de cultura líquidos para a produção de Metarhizium anisopliae (IBCB 425) e Beauveria bassiana (IBCB 66), bem como a tolerância desses propágulos à ação dos raios ultravioleta e à temperatura. Foram avaliados 12 tratamentos compostos pelas combinações entre carbono e nitrogênio. Para determinar o efeito da radiação ultravioleta, placas com blastósporos foram expostas à radiação por 25 e 50 segundos. Para conhecer o efeito da temperatura, os blastósporos de meios de cultura foram expostos a temperaturas de 20, 25, 30 e 35°. Para os experimentos de virulência, as lagartas Diatraea saccharalis foram pulverizadas com 2mL de suspensão fúngica, com o auxílio de uma torre de Potter. Os melhores meios para M. anisopliae são 16,00 g (carbono) + 7,00 g (nitrogênio) e 14,40 g (carbono) + 7,00 g (nitrogênio), e para B. bassiana, 20,00 g (carbono) + 6,30 g (nitrogênio) e 20,00 g (carbono) + 7,00 g (nitrogênio). Quanto mais tempo de exposição à radiação ultravioleta, menor o número de colônias. Aos 35°, há uma diminuição signifcativa na formação de colônias. Os propágulos produzidos pelos fungos são patogênicos a D. saccharalis.(AU)

Structural studies of the exopolysaccharide produced by a submerged culture of entomopathogenic fungus Metarhizium anisopliae / Estudio estructural del exopolisacárido producido por un cultivo sumergido del hongo entomopatogénico Metarhizium anisopliae

The exopolysaccharide (EPS) separated from the entomopathogenic fungus Metarhizium anisopliae was determined by gel permeation chromatography to be homogeneous. The high-performance anion-exchange chromatography with pulsed-amperometric detection (HPAE-PAD) showed a content of monosaccharides D-galactosamine and D-fucose at a molar ratio of about 2:1. The results obtained from Fourier transform-infrared spectroscopy (FT-IR) and second derivative FT-IR spectrum confirmed the proposed structure.

El exopolisacárido (EPS) separado desde el hongo entomopatogénico Metarhizium anisopliae determinado por cromatografía de exclusión en gel ser homogéneo. La cromatografía iónica de alto rendimiento con detección de pulso amperométrico (HPAE-PAD) mostró un contenido de monosacáridos D-galactosamina y D-fucosa en una relación molar de alrededor de 2:1. Los resultados obtenidos desde la espectroscopía infrarroja con transformada de Fourier (FT-IR) y la segunda derivada del espectro FT-IR confirmaron la estructura propuesta.

Patogenicidade de isolados de fungos entomopatogênicos à Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) / Patogenicity of entomopathogenic fungi to Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae)

Este trabalho teve como objetivo avaliar a patogenicidade, em condições de laboratório, de 49 isolados dos fungos entomopatogênicos das espécies Beauveria bassiana e Metarhizium anisopliae, contra lagartas de Spodoptera frugiperda. Lagartas de 3o ínstar foram imersas em suspensões de 1 × 109 conídios/mL de cada um dos isolados por 10 segundos. Posteriormente, foram individualizadas em placas de Petri com dieta artificial, sendo incubadas a 26 ± 1°C e 12 horas de fotofase, e avaliadas, diariamente, durante dez dias. Os isolados de B. bassian amostraram-se mais eficientes, com destaque para o Unioeste 26, que provocou 44,5% de mortalidade e gerou a maior quantidade de conídios nos testes de produção do fungo. Apesar da porcentagem de mortalidade baixa, tais estudos evidenciam potencial dos fungos entomopatogênicos como promissores agentes de controle de S. frugiperda.(AU)

This study aimed to evaluate the pathogenicity, under laboratory conditions, of 49 isolates of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae against Spodoptera frugiperda larvae. Insects (3rd instar caterpillars) were immersed in 1 × 109 conidia/mL suspensions of each of the isolates for 10 seconds. After that, caterpillars were individually placed in Petri dishes with artificial diet and incubated at 26 ± 1°C and 12 hours of photophase, and they were evaluated daily for ten days. The B. bassiana isolates were more efficient, mainly Unioeste 26 strain, which reached 44.5% mortality and generated the greatest number of conidia in production testing of the fungus. Despite the low mortality percentage, these studies showed the potential of entomopathogenic fungi as promising agents for the control of S. frugiperda.(AU)

Rendimento de conídios e germinação de diferentes isolados de Metarhizium anisopliae (Metsch.) Sorok. (Ascomycota: Clavicipitaceae) cultivados em arroz / Yield of conidia and germination of different isolates of Metarhizium anisopliae (Metsch) Sorok. (Ascomycota: Clavicipitaceae) grown on rice

O objetivo desta pesquisa foi determinar a produção e a viabilidade de diferentes isolados de Metarhizium anisopliae em arroz. Foram colocados em sacos de polipropileno medindo 35 cm de comprimento e 22 cm de largura 100 g de arroz pré-cozido, imediatamente autoclavados a 120 °C, por 25 minutos. Após o resfriamento do arroz, inoculou-se 1 mL de uma suspensão contendo 1,0 x 109 conídios/mL de cada isolado testado, sendo acondicionados em câmara climatizada à temperatura de 25 ± 1°C, 70 ± 10% de UR e fotofase de 12 horas de incubação por 10 dias. Decorrido esse período, o arroz + fungo foi acondicionado em bandejas plásticas para promover a conidiogênese do fungo, dentro da câmara climática a 25 ± 1°C, 70 ± 10% de UR e fotofase de 12 horas. As bandejas ficaram empilhadas por quatro dias, cruzando-as por mais quatro dias. O isolado IBCB 425 foi o que mais produziu conídios em arroz com 1,82 x 109 conídios/g de arroz pré-cozido pelo método de bandeja. Com relação à viabilidade dos isolados, o IBCB 425 também apresentou maior capacidade de germinação dos conídios, com 94,84%.(AU)

In order to determine the production and viability of different isolates of Metarhizium anisopliae, 100 g of pre-cooked rice were placed in 35 cm long and 22 cm wide polypropylene bags, being immediately autoclaved at 120°C for 25 minutes. After cooling down, the rice was inoculated with 1 mL of a suspension containing 1,0 x 109 conidia/mL, packed in an incubator at 25 ± 1°C, 70 ± 10% RH and 12 hours of photophase. Plastic bags containing rice + fungus were incubated for 10 days. After this period, rice + fungus was packed in plastic trays to promote fungal conidiogenesis, within the climatic chamber at 2 5 ±1°C, 70 ± 10% RH and 12 hours of photophase. The trays were stacked for four days, being crossed for another four days. The IBCB 425 isolate was the most produced conidia on rice with 1.82 x 109 conidia/g of pre-cooked rice by the tray method. Regarding the viability of the isolates, IBCB 425 also showed a higher germination of conidia, with 94.84%.(AU)

Virulence of Entomopathogenic Fungi Metarhizium anisopliae and Paecilomyces fumosoroseus for the Microbial Control of Spodoptera exigua

The beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) is difficult to control using chemical insecticides because of the development of insecticide resistance. Several pest control agents are used to control the beet armyworm. Entomopathogenic fungi are one of the candidates for eco-friendly pest control instead of chemical control agents. In this study, among various entomopathogenic fungal strains isolated from soil two isolates were selected as high virulence pathogens against larva of beet armyworm. Control efficacy of fungal conidia was influenced by conidia concentration, temperature, and relative humidity (RH). The isolates Metarhizium anisopliae FT83 showed 100% cumulative mortality against second instar larvae of S. exigua 3 days after treatment at 1 x 10(7) conidia/mL and Paecilomyces fumosoroseus FG340 caused 100% mortality 6 days after treatment at 1 x 10(4) conidia/mL. Both M. anisopliae FT83 and P. fumosoroseus FG340 effectively controlled the moth at 20~30degrees C. M. anisopliae FT83 was significantly affected mortality by RH: mortality was 86.7% at 85% RH and 13.4% at 45% RH. P. fumosoroseus FG340 showed high mortality as 90% at 45% RH and 100% at 75% RH 6 days after conidia treatments. These results suggest that P. fumosoroseus FG340 and M. anisopliae FT83 have high potential to develop as a biocontrol agent against the beet armyworm.