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.

Purpureocillium lilacinum and Trichoderma virens for biological control of trichostrongylid parasites of sheep: an in vitro evaluation / Purpureocillium lilacinum e Trichoderma virens no controle biológico de tricostrongilídeos de ovinos: uma avaliação in vitro

Abstract The study evaluated the ovicidal activity of enzymatic extracts of Purpureocillium lilacinum and Trichoderma virens against trichostrongylid eggs from sheep. Filtered extract (FE) and macerated crude extract (MCE) were prepared from fungal cultures in minimal broth. In the experiment, 100 trichostrongylid eggs, obtained from the feces of naturally infected sheep, were exposed to fungal extracts for 24 and 48 hours/25°C. In the control group, eggs were incubated in minimal broth. The number of L1 larvae was ascertained. Each treatment consisted of four repetitions and the experiment was repeated five times. It was observed that the effect of FE and MCE of P. lilacinum and T. virens on egg hatchability differed from that of the control group. MCE of T. virens and P. lilacinum showed higher ovicidal activity than FE over both periods and at 48 hours of exposure, respectively. From the percentage reductions in hatchability of the eggs, MCE was shown to be superior to FE for both fungi. This study demonstrated the ovicidal potential of these fungi against trichostrongylid eggs. However, further studies are needed in order to identify the molecules responsible for the ovicidal effects, and to evaluate the behavior of fungal extracts in biotic and abiotic interactions.

Resumo O estudo avaliou a atividade ovicida de extratos enzimáticos de Purpureocillium lilacinum e Trichoderma virens sobre ovos de tricostrongilídeos de ovinos. Extrato filtrado (EF) e extrato macerado bruto (EMB) foram preparados a partir de culturas fúngicas em caldo mínimo. No ensaio experimental, 100 ovos de tricostrongilídeos, obtidos de fezes de ovinos naturalmente infectados, foram expostos durante 24 e 48 horas/25ºC aos extratos dos fungos. No grupo controle, os ovos foram incubados em caldo mínimo. O número de larvas L1 foi determinado. Cada tratamento consistiu em quatro repetições e o experimento foi repetido cinco vezes. Observou-se que o efeito ovicida do EF e EMB de P. lilacinum e T. virens diferiu do grupo controle. O EMB de T. virens e P. lilacinum evidenciou atividade ovicida superior ao EF em ambos os períodos avaliados e em 48 horas de exposição, respectivamente. O percentual de redução de eclodibilidade evidenciou que o EMB foi superior ao EF em ambos os fungos. Este estudo demonstra o potencial ovicida desses fungos sobre ovos de tricostrongilídeos. No entanto, estudos adicionais são necessários para identificar as moléculas responsáveis pelo efeito ovicida, bem como avaliar o comportamento dos extratos fúngicos em interações bióticas e abióticas.

Distribution of Rhizosphere and Endosphere Fungi on the First-Class Endangered Plant Cypripedium japonicum

Endangered native plant habitats and populations are rapidly disappearing because of climate and environmental changes. As a representative, the abundance of the first-class endangered wild plant, Cypripedium japonicum, has been rapidly decreasing in Korea. The purpose of this study was to evaluate the distribution of rhizosphere and endophytic fungi on C. japonicum in its native habitat. A total of 440 rhizosphere and 79 endosphere fungi isolates were isolated and identified on the basis of their molecular characteristics. Sixty-five genera and 119 fungi species were identified in this study. The genus Trichoderma showed the highest abundance among both rhizosphere and endosphere fungi. Mortierella, Hypocrea, and Penicillium spp. were also relatively dominant species on C. japonicum. The community structures of rhizosphere and endosphere fungi were similar, but endosphere fungi showed greater diversity.

La bacteria Arthrobacter agilis UMCV2 y diversas aminas inhiben el crecimiento in vitro de hongos destructores de madera / Bacterium Arthrobacter agilis UMCV2 and diverse amines inhibit in vitro growth of wood-decay fungi

El reino Fungi está representado por innumerable cantidad de organismos entre los cuales se encuentran hongos patógenos que deterioran los principales componentes estructurales de la madera, como celulosa, hemicelulosa y lignina. El objetivo de nuestro trabajo fue caracterizar la actividad antifúngica y la producción de diversas aminas de Arthrobacter agilis UMCV2 con acción antagónica sobre hongos xilófagos. Para ello, se aislaron 4 organismos fúngicos (designados en conjunto UMTM) a partir de madera en descomposición en un bosque de pino encino de la comunidad de Cuanajo, Michoacán, México. Dos de ellos presentaron una clara actividad enzimática de celulasas, xilanasas y enzimas accesorias óxido-reductoras, y fueron identificados como pertenecientes a 2 géneros agresivos para la madera: Hypocrea (aislado UMTM3) y Fusarium (aislado UMTM13). In vitro, las aminas evaluadas mostraron tener efecto inhibitorio sobre el crecimiento de los UMTM y la dimetilhexadecilamina; uno de estos compuestos mostró un fuerte potencial para ser utilizado como tratamiento preventivo contra el ataque de hongos destructores de madera.

The kingdom Fungi is represented by a large number of organisms, including pathogens that deteriorate the main structural components of wood, such as cellulose, hemicellulose and lignin. The aim of our work was to characterize the antifungal activity in Arthrobacter agilis UMCV2 and diverse amines against wood-decaying fungi. Four fungal organisms (designated as UMTM) were isolated from decaying wood samples obtained from a forest in Cuanajo-Michoacán, México. Two of them showed a clear enzymatic activity of cellulases, xylanases and oxido-reducing enzymes and were identified as Hypocrea (UMTM3 isolate) and Fusarium (UMTM13 isolate). In vitro, the amines showed inhibitory effect against UMTM growth and one of the amines, dimethylhexadecylamine (DMA16), exhibited strong potential as wood preventive treatment, against the attack of decaying fungi.

A novel pullulanase from a fungus Hypocrea jecorina QM9414: production and biochemical characterization.

Pullulanase production from a fungus Hypocrea jecorina QM9414 that produces native extracellular hydrolases having industrial applications was carried out in a shaking flask culture containing 0.5% amylopectin at a pH of 6.50 at 30°C. The enzyme was purified 11-fold by ammonium sulfate fractionation, anion-exchange and gel-filtration chromatographies with a yield of 10.12% and a specific activity of 1.36 ± 0.14 U/mg protein. The molecular mass of pullulanase was estimated to be 130.56 kDa by PAGE and SDS-PAGE, indicating that the native enzyme was a monomer. The optimum pH and temperature for purified enzyme was 6.5 and between 35°-65°C, respectively. The Km values for amylopectin, starch and pullulan as substrates were 10.7, 15.5 and 38.4 mg/mL, respectively. The Vmax values were found to be 3.32, 3.32 and 3.82 ΔA/min for amylopectin, starch and pullulan, respectively. The enzyme was stable at 40-70°C for 30 min, but lost about 33% of its activity at 80°C and about 43% of activity at 90°C and 100°C for the same incubation period. Pullulanase activity was stimulated by CoCl2, NiCl2, KI, NaCl, MgCl2, and LiSO4. The enzyme was slightly inhibited by urea, CaCl2 and b-mercaptoethanol. The enyzmatic characteristics, substrate specificity and the products of hydrolysis indicated that the enzyme was similar to those of type II pullulanases.

Anticancer and antimicrobial activity of mangrove derived fungi Hypocrea lixii VB1 / 中国天然药物

AIM@#Mangrove is one of the oldest living tree species and its leaves are among the most extensively studied botanicals in use today. Scientific research throughout the world has found evidence to support the fact that its foliar extracts have great potential against human microbial pathogens. This study highlights the isolation of foliar fungi from Rhizophora mucronata, Avicenna officialis and Avicenna marina.@*METHOD@#It was isolated in Sabouroud's Dextrose Agar and mass cultivation was done in Sabouroud's Dextrose broth.@*RESULTS@#The ethyl acetate extract showed maximum antibacterial activity which inturn checked for different concentration against bacterial pathogens and anticancer activity for Hep2 and MCF7 cell line in vitro. The DNA was isolated from the fungi and the ITS region of 5.8 s RNA was sequenced and assigned to new species as they are separated from the type strains phylogenetic neighbors by sequence similarities.@*CONCLUSION@#This preliminary screening of fungal endophytes revealed their potential to yield potent bioactive compounds for drug discovery programmes.

Identification and Characterization of Gliocladium viride Isolated from Mushroom Fly Infested Oak Log Beds Used for Shiitake Cultivation

A green mold species that has not previously been reported in Korea was isolated from oak log beds used for shiitake (Lentinula edodes) cultivation that were infested by mushroom flies. In this study, we identify the mold species as Gliocladium viride (an anamorph of Hypocrea lutea) and describe its mycological properties. The fungus was cottony on both potato dextrose agar (PDA) and Czapek yeast extract agar (CYA), but was colored white on PDA and became yellowish green and brown on CYA. Mycelial growth on PDA attained a diameter of 73 mm at 30degrees C after 5 days. The fungus grew faster on malt extract agar (> 80 mm, 5 days at 25degrees C) compared to CYA and PDA (< 68 mm, 5 days at 25degrees C). Penicillate conidiophores of the fungus are hyaline, smooth walled, branching above typically in four stages, and 120~240 microm in length. Club-shaped or slender phialides are formed on the metulae. Conidia of the fungus were ovate and elliptic, yellowish brown and green, and 2.5~3.0 microm x 1.8~2.3 microm in size. Typically, slimy conidia are formed in a mass and colored brown to dark green to almost black. The internal transcribed spacer rDNA and translation elongation factor 1 alpha gene sequences of the fungus isolated here show 99% identity with previously identified G. viride strains.

Fungal genus Hypocrea/Trichoderma: from barcodes to biodiversity / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Hypocrea/Trichoderma is a genus of soil-borne or wood-decaying fungi containing members important to mankind as producers of industrial enzymes and biocontrol agents against plant pathogens, but also as opportunistic pathogens of immunocompromised humans and animals, while others can cause damage to cultivated mushroom. With the recent advent of a reliable, BarCode-aided identification system for all known taxa of Trichoderma and Hypocrea, it became now possible to study some of the biological fundamentals of the diversity in this fungal genus in more detail. In this article, we will therefore review recent progress in (1) the understanding of the geographic distribution of individual taxa; (2) mechanisms of speciation leading to development of mushroom diseases and facultative human mycoses; and (3) the possible correlation of specific traits of secondary metabolism and molecular phylogeny.

Fungi Isolated from Pine Wood Nematode, Its Vector Japanese Pine Sawyer, and the Nematode-Infected Japanese Black Pine Wood in Korea

Pine tree death caused by pine wood nematode (PWN) involves phoretic relationships between PWN and its vector Japanese pine sawyer beetle (JPS). In an effort to understand the diversity of fungi involved in PWN life cycle, a total of 176 fungal isolates were collected from PWNs, adults and larvae of JPS, PWN-diseased Japanese black pine that was cut down in 2005 at Jinju, Korea. Based on microscopic observation and colony morphology, and sequence analysis of the ITS rDNA, the fungal isolates were identified at the level of genus. Three genera including Mucor, Ophiostoma, and Penicillium were identified from PWN. Two genera of Ophiostoma and Penicillium were discovered from JPS larvae. From JPS adult beetles, nine genera of Aspergillus, Gibberella, Hypocrea, Irpex, Leptosphaeria, Ophiostoma, Penicillium, and Plectosphaerella and unknown basidiomycetes were found. Ten genera from PWN-infected wood were confirmed as Bionectria, Botrytis, Camarops, Fusarium, Hypocrea, Nectrtia, Mucor, Ophiostoma, Penicillium, and Trichoderma. Penicillium and Ophiostoma were commonly distributed on PWN and its vector and host. This is first report of the fungi associated with PWN and its vector and host in Korea.

Species concepts and biodiversity in Trichoderma and Hypocrea: from aggregate species to species clusters? / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Trichoderma/Hypocrea is a genus of soil-borne or wood-decaying fungi containing members important to mankind as producers of industrial enzymes and biocontrol agents against plant pathogens, but also as opportunistic pathogens of immunocompromised humans. Species identification, while essential in view of the controversial properties of taxa of this genus, has been problematic by traditional methods. Here we will present a critical survey of the various identification methods in use. In addition, we will present an update on the taxonomy and phylogeny of the 88 taxa (which occur as 14 holomorphs, 49 teleomorphs and 25 anamorphs in nature) of Trichoderma/Hypocrea that have been confirmed by a combination of morphological, physiological and genetic approaches.

Biocontrol Effect of Gliocladium virens G1 and Soil Amendment on Astragal Stem Rot Caused by Rhizoctonia solani

In order to find an environment-friendly method to suppress astragal stem rot caused by the isolates of Rhizoctonia solani AG 1 and AG 4, we tested an antagonistic fungus Gliocladium virens G1 was evaluated as a biocontrol agent and estimated inorganic compounds and organic materials were tested for their effect of the disease suppression. G. virens G1 effectively inhibited mycelial growth in a dual culture and caused mycelial lysis in the culture filtrate. No adverse effect was observed when examined for seed germination and seedling growth. Promoted seedling growth was observed with the seed treatment. Seeds of astragal plant were germinated higher in the sterile soil than the natural soil. Of 14 inorganics tested, alum, aluminum sulfate and calcium oxide significantly suppressed the mycelial growth and sclerotial germination. Milled pine bark and oak sawdust also suppressed the mycelial growth. Soil amended with 1% of G. virens G1 composted with pine bark (w/v) significantly controlled astragal stem rot in the glasshouse experiments.