One new glycoside naphthopyranone from the Yiling cave-derived Metarhizium anisopliae NHC-M3-2 / 药学学报

Seven compounds were isolated from fermentation extract of cave-derived Metarhizium anisopliae NHC-M3-2 by silica gel, semi-preparative HPLC and other chromatographic methods. Their structures were elucidated by UV, IR, MS and NMR methods as 2,3-dehydroindigotide G (1), (-)-regiolone (2), naphtho-γ-pyrone (3), indigotide G (4), indigotide B (5) destruxin A (6) and destruxin B (7). Compound 1 is a new glycoside naphthopyranone compound. The anti-hepatitis B virus (HBV) activity of these compounds was evaluated. The EC50 and CC50 of compound 3 against HBV were 4.5 μmol·L-1 and 92.3 μmol·L-1, respectively. This is the first report of the antiviral activity of compound 3.

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.

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)

Selectivity of different biological products to the egg parasitoid Telenomus remus (Hymenoptera: Platygastridae)

ABSTRACT The selectivity of five entomopathogens and a chemical insecticide (positive control) to pupae and adults of the egg parasitoid Telenomus remus (Nixon) was evaluated in the laboratory under controlled environmental conditions according to protocols established by the International Organization for Biological Control. Baculovirus anticarsia (Baculovirus AEE®), Bacillus thuringiensis var. kurstaki (Thuricide®), Bacillus thuringiensis var. aizawai (Agree®), Beauveria bassiana (Boveril®), Metarhizium anisopliae (Metarril®) and Trichoderma harzianum (Trichodermil®) are harmless to T. remus pupae, and adults. Thus, our results suggest that the insect control strategies applied here are compatible since entomopathogens were classified as harmless to T. remus in most examined cases and therefore facilitate a joint application to control different pests. Bacillus thuringiensis var. kurstaki (Dipel®), despite being classified as slightly harmful in some of the evaluations, can still be considered compatible for use together with T. remus, especially when compared with chemical insecticides such as chlorpyrifos that might be considered harmful to the parasitoid survival.

Infection of silkworm larvae by the entomopathogenic fungus Metarhizium anisopliae / Infecção de lagartas do bicho-da-seda pelo fungo entomopatogênico Metarhizium anisopliae

ABSTRACT: The isolate E9 of Metarhizium anisopliae was used in commercial hybrids of Bombyx mori larvae to evaluate its biological effect. Symptomatological analyses showed typical signs of fungal infection. Histopathology revealed the presence of large numbers of hemocytes in the hemocoel, and on the sixth dpi the bodies of the insects appeared to be colonised by the fungus. The isolate E9 is pathogenic to larvae B. mori and; therefore, death of the insects was caused by the colonization of fungus in the epidermal and mesodermal tissues.

RESUMO: O isolado E9 de Metarhizium anisopliae foi usado em larvas híbridas de Bombyx mori para avaliar seu efeito biológico. A sintomatologia revelou sinais típicos de infecção por fungos. Na histopatologia foi verificado um aumento no número de hemócitos, sendo que, no 6 dpi, todo o corpo do inseto se apresentou colonizado pelo fungo. O isolado E9 é patogênico para lagartas de B. mori, causando sua morte pela colonização dos tecidos de origem epidérmica e mesodérmica.

Virulence evaluation of entomopathogenic fungi to subterranean termites, Globitermes sulphureus (INSECTA: ISOPTERA)

Aims: Subterranean termites, Globitermes sulphureus is one of the significant pests for agricultural crops such as coconut and oil palm, and occasionally attacks building structure in Malaysia. Efforts to control subterranean termite infestations depended heavily on liquid termiticide applications. Natural pathogen of termites such as entomopathogenic fungi (EPF) is a promising alternative to chemical control. The objective of this study was to determine the most virulent EPF such as Metarhizium anisopliae, Beauveria bassiana and Paecilomyces sp. against the subterranean termites, G. sulphureus for the development of bio-insecticide for future use of termite control. Methodology and results: Subterranean termites were collected from mound-building termites found in Universiti Malaysia Terengganu (UMT). Cultivating, harvesting and counting of conidia of EPF were carried out to prepare the desired concentrations for screening test and bioassays, which were 1×103 , 1x105 , 1×107 and control. The pathogenicity testing was observed daily within 1 week under laboratory conditions. Screening showed that M. anisopliae was found to be the most virulent compared to B. bassiana and Paecilomyces sp., achieving 100% mortality within 3-4 days. M. anisopliae was then further tested on termites and it was found that concentration of 1×107 showed the lowest LT50 value, while LC50 of M. anisopliae in 1 day was 2.0151×106 . Conclusion, significance and impact of study: It could be concluded that M. anisopliae is the most virulent EPF against termites and the most effective concentration was ±107 followed by strains of B. bassiana and Paecilomyces sp. However, there is limited field evaluation of EPF against termites in Malaysia. Evaluation on the efficiency of M. anisopliae in the field should be conducted so that its efficacy could be proven and marketed.

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)

Biological Parameters of Euschistus heros (F.) (Heteroptera: Pentatomidae) and its Susceptibility to Entomopathogenic Fungi When Fed on Different Diets

ABSTRACT: The effects of six diets on biological parameters of Euschistus heros (F.) (Heteroptera: Pentatomidae) and its susceptibility to entomopathogenic fungi Beauveria bassiana (Bb) and Metarhizium anisopliae (Ma) were studied. The diets included a standard diet (bean pods+soybeans, peanuts and sunflower) and diets in which the bean pods were combined with only one type of grain, either soybean, peanut, sunflower, pea or sesame seeds. The best performance was that of insects fed on standard diet, which had the shortest development period and 49.5% of egg-to-adult viability. The insects fed the pea diet exhibited less than 30% viability and 11% of adults had wing deformations. The finite rate of increase values for the treatment groups were: standard: 1.091; soybean: 1.077; peanut: 1.071; sunflower: 1.076; pea: 1.058; and sesame: 1.065. The susceptibility to Ma (55%) was greater than Bb (40%) only for insects fed on the sesame diet. Considering the effects of diet on susceptibility to both fungal species comparatively among the diet-treatment groups, the stink bugs fed the restrictive diets were more susceptible compared with those fed the standard diet. We concluded that diet composition affects the biological parameters of E. heros and differentially affects their susceptibility to Bb and Ma fungi.

Evaluación de la eficacia de la CEPA MAF1309® de Metarhizium anisopliae en el control biológico de garrapatas adultas de Rhipicephalus microplus en Tunja, Colombia / Evaluation of efficacy of MaF1309® strain of Metarhizium anisopliae in biological control of adult ticks Rhipicephalus microplus in Tunja, Colombia

La garrapata del ganado Rhipicephalus microplus constituye el ectoparásito de mayor importancia en las ganaderías de regiones tropicales y subtropicales, al producir pérdidas directas por reducción de la producción de carne, leche y deterioro de las pieles así como a la transmisión de agentes patógenos para el ganado bovino tales como Babesia bovis, Babesia bigemina y Anaplasma marginale. El tratamiento químico ha sido efectivo para el control de las garrapatas; sin embargo,  su uso excesivo ha propiciado la selección de individuos resistentes y por tal motivo, se hace necesario el desarrollo de nuevas alternativas de control, tales como el uso de hongos entomopatógenos. El objetivo del presente estudio fue evaluar la eficacia in vitro de M. anisopliae para el control de la fase adulta de R. microplus, en Tunja, Colombia. Se usó la prueba de inmersión de adultas para evaluar la eficacia de la cepa MaF1309® de M. anisopliae a las siguientes concentraciones: 1x10(4), 1x10(6) y 1x10(8) conidias/mL. Se observó que en las garrapatas tratadas a una concentración de 1x10(8) conidias/mL se alcanzó el 100% de mortalidad a los 14 d postratamiento (PT), mientras que las concentraciones 1x10(6) y 1x10(4) conidias/mL alcanzaron el 100% de mortalidad a los días 16 y 20 PT, respectivamente. La mortalidad fue directamente proporcional a la concentración empleada del hongo, siendo las garrapatas tratadas con concentraciones mayores aquellas que alcanzaron una mortalidad en menor tiempo. Estos resultados in vitro son considerados el inicio de información de tipo local proyectados al diseño de programas de control para el departamento de Boyacá.

The livestock tick Rhipicephalus microplus is a high importance ectoparasite in cattle raised at tropical and sub-tropical regions producing direct losses since reduce meat and milk production and cause leather injury besides transmission of pathogenic agents such as Babesia bovis, Babesia bigemina y Anaplasma marginale. The chemical treatment has been effective to control tick  however the excessive use has selected resistant individuals; for that reason is important to develop new alternatives such as entomopathogen fungi to keep controlled those individuals. The objective of this research was to evaluate in vitro efficacy of M. anisopliae to control adult stage of R. microplus in Tunja, Colombia. The immersion test was used to evaluate efficacy of MaF1309® strain of M. anisopliae at concentrations of 1x10(4), 1x10(6) y 1x10(8) conidia/mL. The group of ticks treated with 1x10(8) conidia had 100% of mortality 14 post-treatment days (PT), otherwise 1x10(6) y 1x10(4) conidia/mL concentrations have 100% of mortality at 16 and 20 PT days, respectively. Mortality was directly proportional to fungus concentration used, the ticks treated with high concentrations reach high mortality in less time. Those in vitro results start in local information projected to design control programs for the department of Boyacá.

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

Liquid Bioformulation of Metarhizium Anisopliae is Effective for the Management of Cow Pea Mosaic Disease.

Liquid bioformulation of Metarhizium anisopliae amended with oils and adjuvants was prepared. Six oils viz., sunflower, safflower, soybean, mustard, arachnid and coconut oils at three different concentrations, 0.1, 0.5 and 1.0 per cent and three adjuvants viz., glycerol @ 5, 8 and 10 per cent; tween-80 @ 0.02, 0.04, 0.06 and 1.0 per cent and arachnid oil @ 1, 5 and 10 per cent were tested to see their effect on growth and development of M. anisopliae in the bioformulation. Liquid bioformulation of M. anisopliae amended with Glycerol (10.0%) + Sunflower oil (0.5%) was found significantly effective which showed 84.33 per cent and 94.93 per cent higher surface area covered and biomass production respectively than the control. Efficacy of the bioformulation was tested against cow pea aphid, Aphis craccivora in pot as well as field condition. Liquid formulation of M. anisopliae supplemented with Glycerol (10.0%) + Sunflower oil (0.5%) was found to be significantly effective causing aphid mortality of 80 per cent at 30 days after spraying with protection of secondary spread of cow pea mosaic disease up to 100 per cent and 96.03 per cent in pot and field condition respectively. Spraying liquid formulation of M. anisopliae amended with Glycerol (10.0%) + Sunflower oil (0.5%) at 15 days interval for twice proved to be the best treatment with highest yield of 24.03 t/ha with a cost benefit ratio of 1:7.95 as compared to control (11.20 t/ha).

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.

Biological insect control using Metarhizium anisopliae: morphological, molecular, and ecological aspects / Controle biológico de insetos utilizando Metarhizium anisopliae: aspectos morfológicos, moleculares e ecológicos

Microbial control of insects is based on the rational use of pathogens to maintain environmentally balanced pest population levels, and Metarhizium anisopliae has been the most studied and most utilized fungal species for that purpose. The natural genetic variability of entomopathogenic fungi is considered one of the principal advantages of microbial insect control. The inter- and intraspecific variability and the genetic diversity and population structures of Metarhizium and other entomopathogenic fungi have been examined using ITS-RFLP, ISSR, and ISSP molecular markers. The persistence of M. anisopliae in the soil and its possible effects on the structures of resident microbial communities must be considered when selecting isolates for biological insect control.

O controle microbiano consiste na utilização racional de patógenos, visando à manutenção da população de insetos em equilíbrio no ambiente. Metarhizium anisopliae é a espécie mais estudada e utilizada no controle biológico de insetos. A variabilidade genética dos fungos entomopatogênicos pode ser considerada uma das principais vantagens no controle microbiano de insetos e pode ser detectada por meio de marcadores moleculares, como ITS-RFLP, ISSR e ISSP. Esses marcadores são usados para a caracterização inter e intraespecífica de Metarhizium e outros fungos entomopatogênicos e poderão auxiliar na compreensão da diversidade genética e da estrutura das populações destes fungos. A persistência de M. anisopliae no solo e seu possível efeito na estrutura da comunidade microbiana deste solo são características importantes e pouco estudadas, que devem ser consideradas no processo de seleção de isolados para o controle biológico de insetos.

Changes in Thiol Content, Glutathione and Hydrogen Peroxide Levels as a Measure of Oxidative Stress in Periplaneta Americana Treated with Metarhizium Anisopliae Spp.

Periplaneta americana (American cockroach) treated with conidia of entomopathogenic fugal isolates of Metarhizium anisopliae revealed a decrease in the thiol content and an increment in the levels of oxidized glutathione as well as hydrogen peroxide (H2O2). T-SH levels decreased to 48% and 50% at 24 hours and 48 hours post treatment respectively at LC90 of M20 isolate. Low virulent isolate M48, on the other hand, recorded a decrease of 76% and 78% in the T-SH levels at 24 and 48 hours post treatment respectively with LC30 treatment. Remarkable increase of 200% in the levels of H2O2 recorded at LC90 of M20 and 102% increase at LC30 of M19 was remarkable and focuses on the extent of oxidative stress induced by the fungal infection. Dynamics in the levels of GSH, GSSG, H2O2 and thiols in the insects treated with different fungal isolates in a time and dose dependant manner reveals oxidative stress induced by the fungal infection and the information would facilitate to explore the antioxidant defense system in augmented resistance or susceptibility of cockroach against entomopathogenic fungal conidia.

In-Vitro Evaluation of the Entomopathogenic Fungal Isolates of Metarhizium Anisopliae for Compatibility with Pesticides, Fungicides and Botanicals.

Isolates of the entomopathogenic fungus Metarhizium anisopliae were tested for their compatibility with insecticides, fungicides and botanical pesticides, which are being used in the field, as a prerequisite for developing as mycopesticides and their use in IPM programmes. Three concentrations (0.1X, 0.5X and 1X) of each chemical were evaluated in the laboratory based on the recommended dose for field application by food poison technique. Variation in vegetative growth and sporulation of M. anisopliae appeared to be related to the chemical nature of the formulations, its concentration and the fungal isolates in study. M19 and M48 isolates showed compatibility with imidacloprid at 0.5X and 0.1X and with fungicide sulphur at all the concentrations tested. All the four botanicals tested were found to be compatible to all the four fungal isolates and neem gold displayed maximum tolerance, at all the concentrations. M19 displayed an enhancement in the vegetative growth with imidacloprid (2%) and HIT (2-18%). 2% increase in the spore output was also recorded by M19 with chloropyrifos and sulphur. M19 and M48 isolates demonstrated compatibility with pesticides, fungicides and botanicals as well as with a cockroach management pesticide, HIT. The two isolates of M. anisopliae tested emerged as prospecting candidates for use as mycopesticide component in the combined application with pesticides like imidachloprid and fungicide, sulphur as well as botanicals in the IPM programmes.

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.

Distribuição volumétrica de calda contendo Metarhizium anisopliae / Volumetric distribution of spray containing Metarhizium anisopliae

O controle microbiano de Mahanarva fimbriolata é de suma importância para o manejo dessa praga em cana-de-açúcar, porém carece de melhorias quanto à tecnologia de aplicação. O trabalho teve como objetivo definir um modelo de bico de pulverização adequado para o controle de M. fimbriolata, com base no padrão de distribuição de calda aspergida e espaçamento entre bicos. Utilizaram-se os bicos TF4, TTI e AIUB11004VS e caldas a base de fungo Metarhizium anisopliae sem adjuvante e com Agral a 2% e 4%. Avaliou-se o espaçamento entre bicos baseado na construção de curvas de deposição, considerando CV máximo de 10%; ângulo de abertura e vazão dos bicos. Os maiores espaçamentos foram de 85cm em calda com 2% de adjuvante para o bico TF4, 70cm para AIUB11004VS sem adição de adjuvante e 55cm para o bico TTI, independente da calda. Em relação ao ângulo de abertura, houve apenas diferença entre os bicos testados em uma mesma calda. A vazão foi maior para a calda com 4% de adjuvante para os bicos TF4 e TTI, sendo que AIUB11004VS apresentou menores vazões em relação aos outros modelos para calda com 2% e 4% de adjuvante. Conclui-se que o modelo AIUB11004VS é uma importante ferramenta operacional, visando o controle de M. fimbriolata, por apresentar menor consumo de calda e bons resultados de distância entre bicos.

The microbial control of Mahanarva fimbriolata is very important to management of this pest in sugarcane crops, however lacks improvement in spraying technology. The work aimed to evaluate a nozzle model adequate to control M. fimbriolata, based on pattern of spray distribution and the space between nozzles. TF4, TTI and AIUB11004VS nozzles containing the fungus Metarhizium anisopliae without adjuvant, with Agral 2% and 4% were used in the assay. We evaluated spacing between nozzles by constructing deposition curves considering maximum CV of 10%, spray angle and nozzle flow for different sprays. The larger nozzle spacings were 85cm for spray with 2% of adjuvant and TF4 nozzle, 70cm for AIUB11004VS without adjuvant addition and 55cm TTI nozzle, independent of spray used. Despite the spray angle, we observed difference between the nozzles to the same spray. The flow was larger for spray with adjuvant at 4%, to TF4 and TTI, while AIUB11004VS showed minor flows when compared to the others nozzles using 2% and 4% of adjuvant. By presenting good results of distance between nozzles and minor spray consumption, AIUB11004VS is a good tool for control of M. fimbriolata.