Compatibilidad de un bioinsecticida a base de Beauveria bassiana con agroquímicos: efecto en la germinación y la actividad insecticida sobre Diatraea saccharalis

Los barrenadores del tallo del género de Diatraea spp. (Lepidoptera:Crambidae) han sido reconocidos por ser la plaga de mayor impacto en el cultivo de caña de azúcar, en el que causan importantes pérdidas económicas. Para su control, una cepa colombiana de Beauveria bassiana (Bals.) Vuill (aislamiento Bv062) que demostró alta virulencia frente a larvas de D. saccharalis (Fabricius), fue utilizada para el desarrollo de un bioinsecticida formulado como un concentrado emulsionable (CE). Con el fin de integrar este bioinsecticida en un esquema de manejo integrado de plagas, es necesario evaluar su compatibilidad con los agroquímicos utilizados comúnmente en el cultivo de caña de azúcar. Para tal fin se evaluó el efecto in vitro de 11 agroquímicos (fungicidas, insecticidas y herbicidas) sobre la germinación y actividad insecticida de conidios formulados. Inicialmente el CE y los agroquímicos fueron ajustados al doble de la concentración recomendada y mezclados en proporción 1:1, se incubaron durante dos horas, y se determinó el porcentaje de germinación. Los conidios expuestos a los agroquímicos fueron también purificados y utilizados para un bioen-sayo frente a larvas de segundo instar de D. saccharalis. El CE resultó ser compatible con los herbicidas e insecticidas evaluados, manteniendo una germinación y eficacia superior al 80%. Por el contrario, el CE fue incompatible con todos los fungicidas evaua-dos. Estos resultados permiten sugerir recomendaciones de aplicación del bioplaguicida a base de B. bassiana Bv062, junto con insecticidas y herbicidas empleados en el cultivo de caña, pero se recomienda no aplicarlo de manera simultánea con fungicidas como el benomil y del grupo de triazoles. Se recomienda validar estos resultados en condiciones de campo.

Sugarcane stem borers of the genus Diatraea spp. (Lepidoptera:Crambidae) have been recognized as the pest of the greatest impact on sugarcane crops causing significant economic losses. For its control, a colombian strain of Beauveria bassiana (Bals.) Vuill (code Bv062) that showed a high virulence against D. saccharalis (Fabricius) larvae was used for a biopesticide development, formulated as emulsifiable concentrate (EC). In order to include the EC into an integrated pest management (IPM) scheme, it is necessary to evaluate its compatibility with the agrochemicals commonly used in sugarcane crops. For this purpose, the in vitro effect of 11 agrochemicals (fungicides, insecticides and herbicides) on the germination and insecticidal activity of Bv062 formulated conidia was evaluated. Initially, the EC and agrochemicals were adjusted to double the recommended concentration and mixed in a 1:1 ratio. The mixes were incubated for two hours, and the germination percentage was calculated. The conidia exposed to the agrochemicals were also purified and used for a bioassay against second instar larvae of D. saccharalis. The EC was compatible with the herbicides and insecticides, maintaining germination and efficacy higher than 80%. In contrast, EC was incompatible with all the fungicides evaluated. These results allow us to suggest recommendations for the application of the biopesticide based on B. bassiana Bv062 with insecticides and herbicides used in sugarcane crops, but it is not recommended to apply it simultaneously with fungicides such as benomyl and the triazole group. It is recommended to validate these results under field conditions.

Biocontrol potential of Beauveria bassiana and Metarhizium anisopliae isolates from Turkey against Hyphantria cunea (Drury) (Lepidoptera: arctiidae) Larvae under laboratory and field conditions

Hyphantria cunea is one of the most important pest insects causing significant damage in many plant species. The present study aimed to evaluate the insecticidal activity of Turkey isolates of Beauveria bassiana and Metarhizium anisopliae against H. cunea larvae under laboratory and field conditions. B. bassiana isolates YK16, YK23 and YK26, and M. anisopliae isolates YK41 and YK45 were sprayed onto the larvae of H. cunea at the respective doses of 1x105 and 1x106 conidia mL-1 and monitored for seven days. Fungal isolates, bio-insecticide Bacillus thuringiensis and chemical insecticide diflubenzuron were used in field studies. Trials were carried out in five replications. All B. bassiana isolates caused 100% mortality on first instar larvae in laboratory trials. Mortality ratios ranged from 100 to 96% on second instar larvae. M. anisopliae isolates YK45 and YK41 caused 88 and 84%, and 81.33 and 77.11% mortalities for the first and second instar larvae, respectively. The mortality rates fluctuated between 91.78-72.89% for B. bassiana on third instar larvae in laboratory conditions. However, M. anisopliae isolates YK45 and YK41 caused 77.11 and 60.22% mortality on third instar larvae, respectively. In the field trials, B. bassiana YK23 displayed promising insecticidal activity with 80.60% mortality on second instar larvae of H. cunea. Other isolates as well caused mortalities ranging from 60.77 to 49.55%. The results revealed that some isolates of B. bassiana and M. anisopliae have potential to control H. cunea larvae. However, additional detailed studies need to be carried out to increase their effectiveness in field conditions.

Identification of entomopathogenic fungi Metarhizium anisopliae and Purpureocillium lilacinum from oil palm plantation soils in Universiti Putra Malaysia

Aims@#Insect pests are one of the major constraints of oil palm production in Malaysia. However, synthetic chemical insecticides are the most common method for its control, despite their negative effects on non-target organisms and the development of resistance. Therefore, the present study is aimed to identify novel indigenous isolates of entomopathogenic fungi (EPF) in oil palm soil as part of integrated pest management (IPM) of oil palm insect pests.@*Methodology and results@#The potential of EPF were isolated from the soil collected from the oil palm plantation in UPM using a mealworm beetle larva (Tenebrio molitor) as an insect bait. Seven Metarhizium anisopliae and two Purpureocillium lilacinum isolates were identified by morphological characterization (macroscopic and microscopic observation) and molecular identification using internal transcribed spacer (ITS) region (ITS region amplification).@*Conclusion, significance and impact of study@#To our knowledge, this is the first time P. lilacinum has been found in the soil of an oil palm plantation or any other host in Malaysia. Furthermore, both of the isolates M. anisopliae and P. lilacinum may be potentially considered as biological control candidates for major insect pests in oil palm.

Dual effects of entomopathogenic fungi on control of the pest Lobesia botrana and the pathogenic fungus Eutypella microtheca on grapevine

BACKGROUND: Entomopathogenic fungi (EPF) are the natural enemies of insect pests. Nevertheless, research on the use of EPF for simultaneous prevention of pest and disease agents on the same crop is limited. In this study, we explored the potential dual effects of three strains of the EPF Metarhizium anisopliae on the control of detrimental agents of Vitis vinifera L., including different developmental stages (larvae, pupae, and adult) of the insect pest Lobesia botrana and the phytopathogenic fungus Eutypella microtheca. METHODS: Laboratory pathogenicity trials were performed to examine the effects of the three M. anisopliae strains on the mortality rate of L. botrana. In addition, field trials were conducted to assess the biocontrol potential of one selected M. anisopliae strain on the larval stage of L. botrana. Moreover, inhibitory effects of the three EPF strains on E. microtheca growth were examined in vitro. RESULTS: All the M. anisopliae strains were highly effective, killing all stages of L. botrana as well as inhibiting the growth of E. microtheca. The in vitro mortality of larvae treated with the strains was over 75%, whereas that of treated pupae and adults was over 85%. The three EPF strains showed similar efficacy against larvae and adult stages; never-theless, pupal mortality was observed to be strain dependent. Mortality of L. botrana larvae ranged from 64 to 91% at field conditions. Inhibition of E. microtheca growth reached 50% in comparison to the control. CONCLUSIONS: Our study showed that M. anisopliae strains were highly effective in ensuring control of two different detrimental agents of V. vinifera L., providing new evidence to support the dual effects of entomopathogenic fungi.

Conidia production of Isaria fumosorosea (Hypocreales: Cordycipitaceae) in organic substrates through two propagation techniques / Produção de conídios de Isaria fumosorosea (Hypocreales: Cordycipitaceae) en substratos orgânicos atraves de duas técnicas de propagação

In the last decades, the use of entomopathogenic fungi for the control of pest insects has increased globally, however a key step to achieve the successful application of fungal propagules as biocontrol agents depends of various factors, inoculum production being one of the essential stages for these microorganisms to be used in pest management programs. The objective of this study was to evaluate theproduction of conidia by solid fermentation and biphasic culture in different sporulation matrices for four native isolates and a strain of Isaria fumosorosea that have been shown potential for the control of various insects at the laboratory level. The experimental design was completely randomized, two propagation methods were used with six treatments and ten repetitions per treatment; the data were analyzed using an analysis of variance and the means were compared using the Tukey test (p≤ 0.05). In the production by solid fermentation the minimum value of 5.30 × 105 conidia g-1 was registered with the pericarp of peanut and the maximum of 2.35 × 107 conidia g-1 in the corn grain; in the biphasic culture the minimum of 7.60 × 105 conidia g-1 was observed in birdseed and the maximum of 2.07 × 107 conidia g-1 in rice. The differences were significant (p ≤0.05) in the production by method and by substrate, in solid fermentation 6.84 × 106 conidia g-1 and in biphasic culture 8.85 × 106 conidia g-1. In the production by substrate, the rice showed 1.75 × 107 conidia g-1 and the lesser canary seed concentration (7.80 × 105 conidia g-1). The average production per isolate and / or strain was of the order of 106 and significant difference was registered (p ≤ 0.05) among the fungi, the isolate HIB-9 showed concentration in average higher in the production (7.90 × 106 conidia g-1) and the isolated HIB-19 was the least effective, with only 1.08 × 106 conidia g-1. The results obtained show marked differences between the isolates with respect to their capacity to use the different substrates used as propagation media to obtain conidia of I. fumosorosea.

Nas últimas décadas, o uso de fungos entomopatogênicos para o controle de insetos-pragatem aumentado globalmente, porém um passo fundamental para o sucesso da aplicação de propágulos fúngicos como agentes de biocontrole depende de varios fatores, sendo a produção de inóculo um dos estágios essenciais para esses insetos microorganismos a serem usados em programas de controle de pragas. O objetivo deste trabalho foi avaliar a produção de conídios por meio de fermentação sólida e cultura bifásica emdiferentes matrizes de esporulação para quatro isolados nativos e um alinhagem de Isaria fumosorosea que tem demonstrado potencial para o controle de varios insetos em nível laboratorial. O delineamento experimental foi inteiramente casualizado, foram utilizados dois métodos de propagação com seis tratamentos e dezrepetições por tratamento; os dados foraman alisados por meio de análise de variância e as médias comparadas pelo teste de Tukey (p ≤ 0.05). Naprodução por fermentação sólida foi registrado o valor mínimo de 5.30 × 105 conídios g-1 com o pericarpo de amendoim e o máximo de 2.35 × 107 conídios g-1 no grão de milho; na cultura bifásica, o mínimo de 7.60 × 105 conídios g-1 foi observado emsementes de aves e o máximo de 2.07 × 107conídios g-1em arroz. As diferençasforam significativas (p ≤ 0.05) na produção pelo método e pelo substrato, na fermentação sólida 6.84 × 106 conídios g-1 e na cultura bifásica 8.85 × 106 conídios g-1. Naprodução por substrato, o arroz apresentou 1.75 × 107conídios g-1 e a menor concentração de sementes de canário (7.80 × 105conídios g-1). A produção média por isolado e / ou cepa foi da ordem de 106 e registrou-se diferença significativa (p ≤ 0.05) entre os fungos, o isolado HIB-9 apresentou concentração em média maior na produção (7.90 × 106conídios g-1) e o isolado HIB-19 foi o menos efetivo, com apenas 1.08 × 106 conídios g-1. Os resultados obtidos mostram diferenças marcantes entre os isolados quanto à capacidade de utilização dos diferentes substratos utilizados como meio de propagação para obtenção de conídios de I. fumosorosea.

AMAZONIAN ISOLATES OF Metarhizium ARE EFFECTIVE FOR KILLING Bactrocera carambolae (DIPTERA: TEPHRITIDAE) / Aislados Amazónicos de Metarhizium son efectivos para matar Bactrocera carambolae (Diptera: Tephritidae)

ABSTRACT Bactrocera carambolae Drew & Hancock is a quarantine pest present in Brazil and is the main phytosanitary barrier for the export of fresh fruits from that country. In this work, we evaluated the effect of Amazonian isolates of Metarhizium on kill larvae, pupae, and adults of B. carambolae in soil applications. The mortality of larvae and pupae in sterile soil treated with Metarhizium anisopliae was 70 %. In addition, 100 % of the adults that emerged from this substrate died up to five days later. This isolate caused the mortality of larvae and pupae in non-sterile soil, but its effect was more evident after adult emergence (70 % mortality up to ten days after emergence). Metarhizium robertsii was effective in killing larvae and pupae of the pest, especially in non-sterile soil, and caused a 60 % reduction in adult survival 50 days after emergence. These results indicated the possible use of these isolates for the control of B. carambolae, suggesting the possibility of using this biocontrol mode as another alternative in controlling this quarantine pest.

RESUMEN Bactrocera carambolae Drew & Hancock es una plaga cuarentenaria presente en Brasil y es la principal barrera fitosanitaria para la exportación de frutas frescas de este país. En este trabajo, evaluamos el efecto de los aislados amazónicos de Metarhizium para matar larvas, pupas y adultos de B. carambolae en aplicaciones de suelo. La mortalidad de larvas y pupas en suelo estéril tratado con Metarhizium anisopliae fue del 70 %. Además, el 100 % de los adultos que emergieron de este sustrato murieron hasta cinco días después. Este aislado causó la mortalidad de larvas y pupas en suelo no estéril, pero su efecto fue más evidente después de la emergencia del adulto (70 % de mortalidad hasta diez días después de la emergencia). Metarhizium robertsii fue eficaz para matar larvas y pupas de la plaga, especialmente en suelos no estériles, y causó una reducción del 60 % en la supervivencia de los adultos 50 días después de la emergencia. Estos resultados indican que es posible el uso de estos aislados para el control de B. carambolae, lo que sugiere la posibilidad de utilizar este modo de control biológico como otra alternativa para controlar esta plaga cuarentenaria.

Spores of Beauveria bassiana and Trichoderma lignorum as a bioinsecticide for the control of Atta cephalotes

Background: The leafcutter ant (Atta cephalotes) is associated with losses in the agricultural sector, due to its defoliating activity; for its control, biological, mechanical and chemical methods have been developed, the latter associated with adverse effects on human and environmental health. This research validated in the field for the control of the leafcutter ant (A. cephalotes) using a mixture of Beauveria bassiana and Trichoderma lignorum spores. Methods: The effectiveness from the combination of spores of B. bassiana and T lignorum with an initial concentration of 2 × 109 spores/ml, in the following proportions of B. bassiana and T. lignorum, A (1:1), of each fungus. It was evaluated within the university campus, comparing it with two commercial formulations, Mycotrol (B. bassiana) and Mycobac (T. lignorum). Additionally, this formulation was evaluated in 49 nests distributed 16 in 14 locations in Colombia. The formulation application was carried out by direct application, using a pump at a speed of 10 ml/m2. The effectiveness was estimated from the reduction of the flow of ants, evaluating the statistically significant differences using the ANOVA and Tukey-test. Results: Effective control of 90% of the nests was observed in the field phase in 60 days, except in nests with areas > 50 m2 that were located in regions with high rainfall (annual average precipitation above 7000 mm), such as Buenaventura. Conclusions: In this work, it was demonstrated that the combination of B. bassiana and T. lignorum spores represent a viable alternative for the control of the leafcutter ant, in which the effectiveness is related to several factors, including the size of the nest and the rainfall in the area.

Tobacco Growth Promotion by the Entomopathogenic Fungus, Isaria javanica pf185

Isaria javanica pf185 is an important entomopathogenic fungus with potential for use as an agricultural biocontrol agent. However, the effect of I. javanica pf185 on plant growth is unknown. Enhanced tobacco growth was observed when tobacco roots were exposed to spores, cultures, and fungal cell-free culture supernatants of this fungus. Tobacco seedlings were also exposed to the volatiles of I. javanica pf185 in vitro using I-plates in which the plant and fungus were growing in separate compartments connected only by air space. The length and weight of seedlings, content of leaf chlorophyll, and number of root branches were significantly increased by the fungal volatiles. Heptane, 3-hexanone, 2,4-dimethylhexane, and 2-nonanone were detected, by solid-phase micro-extraction and gas chromatography-mass spectrophotometry, as the key volatile compounds produced by I. javanica pf185. These findings illustrate that I. javanica pf185 can be used to promote plant growth, and also as a biocontrol agent of insect and plant diseases. Further studies are necessary to elucidate the mechanisms by which I. javanica pf185 promotes plant growth.

Potential of endophytic fungi as biocontrol agents of Duponchelia fovealis (Zeller) (Lepidoptera: Crambidae) / Fungos endofíticos para o biocontrole de Duponchelia fovealis (Zeller) (Lepidoptera: Crambidae)

Abstract This study reports the first assessment of endophytic fungi isolated from strawberry leaves and selection of isolates for the control of Duponchelia fovealis, a new pest of strawberries. A total of 400 strawberry leaves of the cultivar 'Albion' were collected in four commercial farms. Leaves were disinfected, cut in fragments, and placed on Petri dishes containing potato dextrose agar media with tetracycline and incubated for 30 days. Following this time, 517 fungal colonies were isolated, and thirteen genera were identified: Cladosporium, Aspergillus, Nigrospora, Fusarium, Trichoderma, Chaetomium, Alternaria, Paecilomyces, Penicillium, Ulocladium, Bipolaris, Diaporthe, and Phoma. Eight isolates belonging to the genera Aspergillus, Diaporthe, Paecilomyces, and Cladosporium were selected for pathogenicity bioassays against third instar larvae of D. fovealis. Isolates of Paecilomyces induced the highest mortality rates.

Resumo Este trabalho apresenta a primeira contribuição no isolamento de fungos endofíticos de folhas de morangueiro e na seleção de isolados para controle de Duponchelia fovealis, uma nova praga do morangueiro. Foram coletadas 400 folhas da cultivar 'Albion' em quatro lavouras comerciais de morangueiro. As folhas foram desinfetadas, cortadas em fragmentos e depositadas em placas de Petri contendo Ágar Batata Dextrose como meio, em conjunto com tetraciclina e incubados durante 30 dias. Um total de 517 colônias fúngicas e treze gêneros foram isolados: Cladosporium, Aspergillus, Nigrospora, Fusarium, Trichoderma, Chaetomium, Alternaria, Paecilomyces, Penicillium, Ulocladium, Bipolaris, Diaporthe e Phoma. Oito isolados pertencentes aos gêneros Aspergillus, Diaporthe, Paecilomyces e Cladosporium foram selecionados para os bioensaios de virulência contra larvas de 3º instar de D. fovealis. Isolados de Paecilomyces causaram as maiores taxas de mortalidade.

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.

Effect of entomopathogens on Africanized Apis mellifera L. (Hymenoptera: Apidae)

ABSTRACT This study aimed to evaluate the effect of commercially used entomopathogens on Africanized Apis mellifera L. (Hymenoptera: Apidae). Four bioassays were performed: 1) pulverized entomopathogens on A. mellifera; 2) entomopathogens sprayed on a smooth surface; 3) entomopathogens sprayed on soy leaves; and 4) entomopathogens mixed with candy paste (sugar syrup). Five treatments were prepared: sterile distilled water (control), distilled water sterilized with Tween® 80 (0.01%), and the commercial entomopathogens Metarhizium anisopliae E9 (1.0 × 109 conidia mL−1), Beauveria bassiana PL63 (1.0 × 108 conidia mL−1) and Bacillus thuringiensis var. kurstaki HD-1 (3.0 × 108 spores mL−1). Each treatment consisted of five repetitions, with 20 workers per repetition, which were stored in a plastic box and, later, in a biological oxygen demand (B.O.D.) incubator (27 ± 2 ºC, RH of 60% ± 10%, 12-h photophase). The mortality of the workers was evaluated from 1 h to 240 h, and the data were analyzed using Bayesian inference. The workers killed by the ingestion of candy paste contaminated with the pathogens (products) were randomly separated and selected for the removal of the midgut. Each midgut was fixed in Bouin's solution and prepared for histology. B. bassiana was verified to reduce the survival of A. mellifera workers in all bioassays. Moreover, M. anisopliae reduced the survival of A. mellifera workers directly sprayed, on a smooth surface and mixed with candy. B. thuringiensis reduced A. mellifera survival on a smooth surface and mixed with candy paste. However, its effects were lower than that observed by B. bassiana. The treatments with the biological products did not induce morphometric alterations in the midgut of A. mellifera.

First record of Ophiocordyceps dipterigena (Ascomycota: Hypocreales: Ophiocordycipitaceae) infecting adults of Melanagromyza sojae (Diptera: Agromyzidae) in Brazil / Primeiro registro de Ophiocordyceps dipterigena (Ascomycota: Hypocreales: Ophiocordycipitaceae) infectando adultos de Melanagromyza sojae (Diptera: Agromyzidae) no Brasil

ABSTRACT: This note is the first report on the infection of adult stage of Melanagromyza sojae Zehntner, 1900 (Diptera: Agromyzidae) by the entomopathogenic fungus Ophiocordyceps dipterigena (Hypocreales: Ophiocordycipitaceae) (Berk. & Broome) G. H. Sung, J.M. Sung, Hywel-Jones & Spatafora, in subtropical Brazil. Entomopathogenic fungi, which attack dipteran insects, are of great scientific and economic importance mainly due to their potential as biological control agents of insect pests. Our findings bring new perspectives on the geographical distribution and host range of Ophiocordyceps dipterigena. We emphasize the need of further studies and research on new biological control agents of agricultural pests such as the soybean stem miner fly.

RESUMO: Esta nota é o primeiro registro da infecção no estágio adulto de Melanagromyza sojae Zehntner, 1900 (Diptera: Agromyzidae) pelo fungo entomopatogênico Ophiocordyceps dipterigena (Hypocreales: Ophiocordycipitaceae) (Berk & Broome) G. H. Sung, J.M. Sung, Hywel-Jones & Spatafora, no Brasil subtropical. Fungos entomopatogênicos, que atacam dípteros, são de importância científica e econômica principalmente devido ao potencial como agentes de controle biológico de pragas de insetos. Este trabalho traz novas perspectivas sobre a distribuição geográfica e variação de hospedeiro de Ophiocordyceps dipterigena. Destacamos a necessidade de ampliar o conhecimento sobre novos agentes de controle biológico de pragas agrícolas como a mosca minadora do caule da soja.

Enzymatic comparison and mortality of Beauveria bassiana against cabbage caterpillar Pieris brassicae LINN

Abstract Beauveria bassiana, an entomopathogenic fungus, is the alternative biocontrol agent exploited against major economic crop pests. Pieris brassicae L. is an emerging pest of the Brassicaceae family. Therefore, in the present study, fungal isolates of Beauveria bassiana, viz. MTCC 2028, MTCC 4495, MTCC 6291, and NBAII-11, were evaluated for their virulence against third instar larvae of P. brassicae. Among all these fungal isolates, maximum mortality (86.66%) was recorded in B. bassiana MTCC 4495 at higher concentration of spores (109 conidia/ml), and the minimum mortality (30.00%) was recorded in B. bassiana MTCC 6291 at a lower concentration (107 conidia/ml) after ten days of treatment. The extracellular cuticle-degrading enzyme activities of fungal isolates were measured. Variability was observed both in the pattern of enzyme secretion and the level of enzyme activities among various fungal isolates. B. bassiana MTCC 4495 recorded the maximum mean chitinase (0.51 U/ml), protease (1.12 U/ml), and lipase activities (1.36 U/ml). The minimum mean chitinase and protease activities (0.37 and 0.91 U/ml, respectively) were recorded in B. bassiana MTCC 6291. The minimum mean lipase activity (1.04 U/ml) was recorded in B. bassiana NBAII-11. Our studies revealed B. bassiana MTCC 4495 as the most pathogenic isolate against P. brassicae, which also recorded maximum extracellular enzyme activities, suggesting the possible roles of extracellular enzymes in the pathogenicity of B. bassiana against P. brassicae.

Pathogenicity of Metarhizium anisopliae isolates on Nezara viridula and Dichelops melacanthus in wheat crop / Patogenicidade de isolados de Metarhizium anisopliae (Metsch) sobre Nezara viridula e Dichelops melacanthus (Hemiptera: Pentatomidae) na cultura do trigo

Bugs such as Nezara viridula (Linnaeus) and Dichelops melacanthus (Dallas) are considered the main insect pests of wheat crop in Brazil. The use of the entomopathogenic fungus Metarhizium anisopliae (Metschnikoff) may be an alternative for the management of these insects in the crop. The objective of this work was to verify the pathogenicity of different isolates of M. anisopliae on adults of N. ­viridula and D. melacanthus under laboratory and greenhouse conditions. In the laboratory, isolates 05RA, 11RA, 08RA and 02RA were obtained from N. viridula and D. melacanthus infested with M. anisopliae. Also, a high pathogenicity (100% of mortality) of both species was recorded in a bioassay of the topical application 8 Days After Application (DAA). However, compared to the other isolates, the 08RA isolate showed the highest pathogenicity in a shorter time interval for N. ­viridula (Mean Time "MT" = 2.8 days) and D. melacanthus (MT = 4.0 days). Under greenhouse conditions, the 08RA isolate provided a mortality of 44.9% (N. viridula) and 35.7% (D. melacanthus) in the same evaluation period. However, at 14 DAA, the mortality was 100% for both species, with the MT values of N. ­viridula and D. ­melacanthus being obtained at 8 days and 10 days, respectively. The fungus M. anisopliae is a promising alternative for the control of adult N. viridula and D. melacanthus in wheat crop.(AU)

Os percevejos Nezaraviridula (Linnaeus) e o Dichelops melacanthus (Dallas) são considerados os principais insetos-pragas na cultura do trigo no Brasil. A utilização do fungo entomopatogênico Metarhiziumanisopliae (Metschnikoff) pode ser uma alternativa para o manejo destes insetos na cultura. O objetivo do trabalho foi verificar a patogenicidade de diferentes isolados de M.anisopliae sobre adultos de N. viridula ou D. ­melacanthus em condições de laboratório e casa de vegetação. Em laboratório, os isolados 05RA, 11RA, 08RA e 02RA provenientes de adultos de N. ­viridula ou D. melacanthus, infestados a campo pelo fungo M. anisopliae, apresentaram elevada patogenicidade (100% de mortalidade) de ambas as espécies em bioensaio de aplicação tópica 8 dias após aplicação (DAA). Entretanto, o isolado 08RA apresentou a maior patogenicidade em um menor intervalo de tempo para N. ­viridula (Tempo Médio "TM" = 2,8 dias) e D. melacanthus (TM = 4,0 dias) em relação aos demais isolados. Em casa de vegetação, o isolado 08RA proporcionou mortalidade de 44,9% (N. ­viridula) e 35,7% (D. ­melacanthus) no mesmo período de avaliação. Contudo, aos 14 DAA, a mortalidade foi de 100% para ambas as espécies, com valores de TM de 8 dias e 10 dias para N. viridula e D. ­melacanthus, respectivamente. O fungo M. anisopliae constitui uma alternativa promissora de controle de adultos de N. ­viridula ou D. ­melacanthus na cultura do trigo.(AU)

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.

White grub (Coleoptera: Melolonthidae) mortality induced by Ophiocordyceps melolonthae / Mortalidade em corós (Coleoptera: Melolonthidae) induzida por Ophiocordyceps melolonthae

ABSTRACT: The occurrence of white grub roots in soybean crops in the South of Brazil has gradually increased. However, there is not information on the biological control of grubs by entomopathogenic fungi. This study aimed to induce infection by Ophiocordyceps melolonthae and analyze longevity in Cyclocephala modesta and Dyscinetus gagates larvae (Coleoptera: Melolonthidae). In the laboratory, Cyclocephala modesta and Dyscinetus gagates had a mortality rate of 85% and 75%, respectively.

RESUMO: A ocorrência de corós-praga de raízes em lavoura de soja, no Sul do Brasil, tem gradualmente aumentado. Entretanto, não há informação sobre controle biológico de corós por fungos entomopatogênicos. Este estudo tem o objetivo de induzir a infecção por Ophiocordyceps melolonthae e analisar a longevidade em larvas de Cyclocephala modesta e Dyscinetus gagates (Coleoptera: Melolonthidae). Em laboratório, Cyclocephala modesta e Dyscinetus gagates apresentaram uma taxa de mortalidade de 85% e 75%, respectivamente.

Potential of endophytic fungi as biocontrol agents of Duponchelia fovealis (Zeller) (Lepidoptera:Crambidae)

Abstract This study reports the first assessment of endophytic fungi isolated from strawberry leaves and selection of isolates for the control of Duponchelia fovealis, a new pest of strawberries. A total of 400 strawberry leaves of the cultivar Albion were collected in four commercial farms. Leaves were disinfected, cut in fragments, and placed on Petri dishes containing potato dextrose agar media with tetracycline and incubated for 30 days. Following this time, 517 fungal colonies were isolated, and thirteen genera were identified: Cladosporium, Aspergillus, Nigrospora, Fusarium, Trichoderma, Chaetomium, Alternaria, Paecilomyces, Penicillium, Ulocladium, Bipolaris, Diaporthe, and Phoma. Eight isolates belonging to the genera Aspergillus, Diaporthe, Paecilomyces, and Cladosporium were selected for pathogenicity bioassays against third instar larvae of D. fovealis. Isolates of Paecilomyces induced the highest mortality rates.

Resumo Este trabalho apresenta a primeira contribuição no isolamento de fungos endofíticos de folhas de morangueiro e na seleção de isolados para controle de Duponchelia fovealis, uma nova praga do morangueiro. Foram coletadas 400 folhas da cultivar 'Albion' em quatro lavouras comerciais de morangueiro. As folhas foram desinfetadas, cortadas em fragmentos e depositadas em placas de Petri contendo Ágar Batata Dextrose como meio, em conjunto com tetraciclina e incubados durante 30 dias. Um total de 517 colônias fúngicas e treze gêneros foram isolados: Cladosporium, Aspergillus, Nigrospora, Fusarium, Trichoderma, Chaetomium, Alternaria, Paecilomyces, Penicillium, Ulocladium, Bipolaris, Diaporthe e Phoma. Oito isolados pertencentes aos gêneros Aspergillus, Diaporthe, Paecilomyces e Cladosporium foram selecionados para os bioensaios de virulência contra larvas de 3º instar de D. fovealis. Isolados de Paecilomyces causaram as maiores taxas de mortalidade.

Infectivity of housefly, Musca domestica (Diptera: Muscidae) to different entomopathogenic fungi

Abstract The housefly Musca domestica is a worldwide insect pest that acts as a vector for many pathogenic diseases in both people and animals. The present study was conducted to evaluate the virulence of different local isolates of Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea on M. domestica using two bioassay techniques: (1) adult immersion and (2) a bait method applied to both larvae and adults. The results showed evidence of a broad range of responses by both stages (larvae and adults) to the tested isolates of B. bassiana, M. anisopliae and I. fumosorosea. These responses were concentration-dependent, with mortality percentages ranging from 53.00% to 96.00%. Because it resulted in lower LC50 values and a shorter lethal time, B. bassiana (Bb-01) proved to be the most virulent isolate against both housefly larvae and adults. Sublethal doses of the tested isolates were also assessed to evaluate their effect on M. domestica fecundity and longevity. The fungal infections reduced housefly survival regardless of their sex and also decreased egg production in females.

Selection and characterization of Beauveria spp. isolates to control the broad mite Polyphagotarsonemus latus (Banks, 1904) (Acari: Tarsonemidae) / Seleção e caracterização de isolados do fungo Beauveria spp. visando ao controle do ácaro branco Polyphagotarsonemus latus (Banks, 1904) (Acari: Tarsonemidae)

Abstract This study was performed under laboratory conditions to identify isolates of the fungus Beauveria spp. that can control Polyphagotarsonemus latus in the greenhouse and field. Thirty Beauveria spp. isolates were tested by spraying 1 mL conidia (1 × 108 conidia/mL) on pepper leaf discs containing 15 mites. Evaluations were performed on the 3rd and 6th day post application by counting the number of dead mites. Vegetative growth and conidial production were measured from the selected isolates, and bioassays were conducted in the greenhouse on bean seedlings in plastic pots. The isolate Unioeste 53 was selected, and a conidial suspension (1 × 108 conidia/mL) was applied with a backpack sprayer. The evaluation consisted of pre- and post-treatment counts of the number of live mites on ten leaflets in both the plots treated with the fungus and control plots, and the same procedure was followed for the field experiment. In the laboratory, the Unioeste 53 isolate resulted in total and confirmed mortality rates of 70% and 57.7%, respectively. In the greenhouse, the population decreased by 76.71% by the 16th day after application. In the field, the population decreased by 66% by the 12th day after application, demonstrating the potential of this fungus for mite management.

Resumo Este trabalho teve por objetivo selecionar isolados do fungo Beauveria bassiana em condições de laboratório com potencial de uso em casa de vegetação e campo no controle do ácaro branco. Foi realizada uma seleção com 30 isolados de Beauveria spp. através de pulverização direta de conídios (1 × 108 conídios/mL) sobre discos foliares de pimenta contendo 15 ácaros. As avaliações foram realizadas no terceiro e sexto dia contando-se o número de mortos, e confirmação do patógeno em câmara úmida. Parâmetros de crescimento vegetativo e produção de conídios foram avaliados. No bioensaio em casa de vegetação foram preparados vasos com plantas de feijão e fez-se a aplicação do Unioeste 53 (1 × 108 conídios/mL), utilizando pulverizador costal, a avaliação constou da contagem prévia e posterior à aplicação do número de ácaros vivos em 10 folíolos, tanto nas parcelas destinadas ao tratamento com fungo, quanto na testemunha. A fase de campo seguiu os mesmos padrões, porém, com área experimental total de 225 m2, com oito parcelas de 10,24 m2, sendo 4 testemunhas e 4 onde foi aplicado o isolado Unioeste 53 seguindo metodologia de aplicação e avaliação já descritas para casa de vegetação. Em laboratório o isolado Unioeste 53 causou mortalidade total de 70% e 57,7% de mortalidade confirmada. Em casa de vegetação, apresentou redução da população de 76,71% 16 dias após aplicação, já em campo, a redução da população foi de 66% após 12 dias da aplicação, demonstrando o potencial do ácaro pelo fungo.

Estabilidade de conídios de Beauveria bassiana (Bals) Vuill acondicionados em diferentes embalagens / Stability of Beauveria bassiana (Bals) Vuill conidia stored in different packagings

Um dos fatores limitantes à utilização em maior quantidade de fungos entomopatogênicos produzidos é a dificuldade na manutenção da viabilidade dos conídios por longos períodos de armazenamento, o que torna importante a necessidade de desenvolvimento de formulações e embalagens que aumentem a vida de prateleira desses micro-organismos. Diante disso, o objetivo deste trabalho foi identificar embalagens que pudessem manter a viabilidade dos conídios por longos períodos de armazenamento em diferentes temperaturas. O trabalho foi desenvolvido no Laboratório de Controle Microbiano de Insetos da Universidade Estadual de Londrina (UEL). Foi utilizada uma mistura granulada (conídios e arroz parboilizado) de B. bassiana (CG 432) acondicionada em 3 tipos de embalagens: Coex, poliéster metalizado e polietileno, que foram armazenadas durante 180 dias em refrigerador (6 ± 2°C) e câmara climatizada (25 ± 1°C). Os parâmetros utilizados para avaliar a influência das embalagens sobre a viabilidade dos conídios foram a porcentagem de germinação e o conteúdo de água. Os conídios acondicionados na embalagem tipo Coex foram os que apresentaram maior porcentagem de germinação em refrigerador e à temperatura ambiente durante todo o período de armazenamento, sendo também a embalagem que proporcionou menor ganho de água pelos conídios durante o armazenamento. Independentemente da embalagem, os conídios armazenados em refrigerador apresentaram maior porcentagem de germinação em relação aos armazenados à temperatura ambiente (25 ± 1°C).(AU)

One of the limiting factors in the use of more substantial amounts of entomopathogenic fungi is the difficulty in maintaining conidial viability for long periods in storage. The need for developing formulations and packaging that would increase the shelf life of these microorganisms is therefore important. Considering this, this study aimed to identify packaging types that would maintain conidial viability for long storage periods at different temperatures. A granular B. bassiana (CG 432) mixture (conidia and parboiled rice) was used, stored in three different packaging types: Coex, metallized polyester, and polyethylene, which were stored in refrigerator (6 ± 2°C) and in an incubator (25 ± 1°C) for 180 days. Germination percentage and water content were the parameters used to evaluate packaging influence on conidial viability. The conidia stored in Coex packaging showed the highest germination percentage in refrigerator and at room temperature during the entire storage period. This packaging also provided the lowest water acquisition by the conidia during storage. Regardless of packaging, the conidia stored in refrigerator showed the highest germination percentage in relation to those stored at room temperature (25 ± 1°C).(AU)