Resumen El control biológico en caña de azúcar en Venezuela se ha logrado establecer como una medida única, con resultados óptimos del uso de parasitoides y entomopatógenos para el control de los taladradores del género Diatraea, y, posteriormente, de la candelilla Aeneolamia spp., ambas plagas claves del cultivo. Para los taladradores, se logró después de liberaciones sistemáticas de la mosca amazónica Lydella minense desde un inicio, a mediados del siglo XX y luego con el establecimiento de la avispa bracónida Cotesia flavipes, a partir de 1987. Para el control biológico de Aeneolamia varia, plaga que implicaba la aplicación frecuente de insecticidas, después de la introducción del uso del hongo entomopatógeno Metarhizium anisopliae desde 1977, se ha establecido como medida eficiente para el control de sus adultos, aunque las ninfas eran menos controladas. Este aspecto se resolvió mediante la introducción del nematodo entomopatógeno Heterorhabditis bacteriophora en 2004, el cual se viene utilizando en forma general, y prácticamente ya se tienen las medidas biológicas para evitar la aplicación de insecticidas. Como ejemplo se tiene a la Azucarera Río Turbio, aunque en las últimas zafras ha tenido una reducción en sus áreas, por diferentes factores. Durante los últimos 58 años, la infestación de Diatraea muestra una reducción de 15 a 3.18 %, con lo cual se salvan 170 552.62 toneladas métricas (TM), con un valor de US$ 49 196 045 y un costo del control biológico de US$ 1 187 189 lo que se traduce en un beneficio-costo de US$ 41.44. Antes del establecimiento del control biológico, Aeneolamia se controlaba exclusivamente con insecticidas químicos, aplicados al follaje para los adultos y al suelo para las ninfas. Simultáneamente desde entonces, se ha utilizado el hongo M. anisopliae producido por empresas nacionales, hasta cubrir 87 000 ha, entre 1986 y 1990. También, desde 2004 se inició la utilización de nematodos entomopatógenos para el control de ninfas de Aeneolamia.

Abstract The biological control in sugarcane in Venezuela has been established as a unique measure, having optimal results of the use of parasitoids and entomopathogens for the control of borers of the genus Diatraea, and later of the candelilla Aeneolamia spp., both key pests of the crop. For borers it was achieved after systematic releases of the Amazonian fly Lydella minense from the middle of the last century and then with the establishment of the braconid wasp Cotesia flavipes from 1987. For the biological control of Aenolamia varia, pest that involved the frequent application of insecticides, after the introduction of the use of the entomopathogenic fungus Metarhizium anisopliae since 1977, has been established as an efficient measure for the control of its adults, however, the nymphs were less controlled. This aspect was resolved through the introduction of the entomopathogenic nematode Heterorhabditis bacteriophora in 2004, which has been widely used as a biological measure to avoid the application of insecticides. As an example, we have the Azucarera Río Turbio, although in the last harvests it has had a reduction in its areas due to different factors. During the last 58 years, Diatraea infestation shows a reduction of 15 to 3.18 %, which saves 170 552.62 metric tons with a value of US$ 49 196 045 and a cost of biological control of US$ 1 187 189 which translates into a benefit-cost of US$ 41.44. Before the establishment of biological control, Aeneolamia was controlled exclusively with chemical insecticides applied to foliage for adults and soil for nymphs. Simultaneously since then, the entomopathogenic fungus M. anisopliae produced by national companies has been used and this entomopathogen was used on 87 000 hectares between 1986 and 1990. Also since 2004, the methodology of using entomopathogenic nematodes to control Aeneolamia nymphs began.

ABSTRACT The horizontal transmission of the entomopathogenic fungus Beauveria bassiana Unioeste 76 strain among adults of Euschistus heros was evaluated in the laboratory and greenhouse using sporulated cadavers and live insects contaminated as a source of inoculate. Adult insects in the laboratory bioassay either received treatments with dry conidia or were sprayed with an oily dispersion formulation, and the treatments were comprised of 10, 30, or 50% of contaminated insects. For the greenhouse bioassay, nylon cages were prepared, with soybean plants grown in pots. A total of 20 healthy brown stink bugs were released along with one or three sporulated cadavers of brown stink bugs, attached to the plant stem. In the bioassay with live insects, 20 stink bugs were released, and the treatments were comprised of 10, 30, or 50% contaminated insects (2:18, 6:14, and 10:10, fungus-treated insects : healthy insects). In all experiments, the control group was comprised only of healthy insects. Dead stink bugs were daily disinfested and transferred to a wet chamber for confirmation of death by fungus. Confirmed mortality was observed, which shows the occurrence of horizontal transmission. Mortality was higher when more contaminated insects were used in the population, in most treatments. The occurrence of transmission in the laboratory and the greenhouse emphasizes the potential of the Unioeste 76 strain as mortality agent and as an agent for maintaining the inoculum potential for the brown stink bug. 7 inoculate formulation 10 30 50 prepared pots 2 stem 218, 218 18, 18 (2:18 614, 614 6 14, 14 6:14 1010, 1010 10:10 fungustreated treated . insects) experiments observed population bug 1 3 5 21 (2:1 61 6:1 101 10:1 (2: 6: 10: (2 (

Resumo Nematoides entomopatogênicos (NEP) das famílias Heterorhabditidae e Steinernematidae são amplamente utilizados no controle de insetos-pragas. No Brasil, os estudos relacionados a caracterização e identificação destes entomopatógenos são recentes e escassos. Nesse sentido, o objetivo deste estudo foi isolar NEP de amostras de solos coletadas em diferentes áreas no estado do Paraná, Brasil. Um isolado Heterorhabditis (UEL 08) detectado em amostra de solo em área de pastagem cultivada com braquiária, localizada em Londrina, Paraná, Brasil (23º34´311´´S, 050º58´298´´W), utilizando o método de “inseto-isca” com lagartas de Galleria mellonella. Para a identificação foram realizados estudos de morfometria e identificação molecular a partir da amplificação da região ITS. Algumas diferenças foram encontradas em termos de morfometria em comparação com a descrição original, entretanto, os dados moleculares demonstraram que a sequência obtida para o isolado UEL 08 é idêntica à sequência de referência de H. amazonensis (DQ665222), com a qual apresentou 100% de similaridade. Os estudos das características biológicas de H. amazonensis (UEL 08) revelaram que o isolado tem dois ciclos de vida distintos, um curto (216 h) e outro longo (288 h), sendo que ocorrem duas gerações em ambos os ciclos. O isolado UEL 08 H. amazonensis foi patogénico e virulento sobre os três hospedeiros avaliados. Notadamente, as larvas de G. mellonella e Alphitobius diaperinus foram consideradas mais susceptíveis do que os adultos do percevejo Dichelops (Diacereus) melacanthus, com percentagens de mortalidade de 100%, 85% e 46% de mortalidade, respectivamente. O ensaio de produção in vivo revelou um rendimento médio diário de 3,4 × 103 juvenis infectantes/g de larva hospedeira.

Abstract Entomopathogenic nematodes (EPNs) from Heterorhabditidae and Steinernematidae families are extensively used to control insect pests. In Brazil, however, relatively few studies have identified and characterized these entomopathogens. The objective of this study was to identify and characterize an EPN isolate obtained from soil samples collected in the state of Paraná, Brazil. An isolate (UEL 08) of Heterorhabditis was detected in a soil sample collected from a pasture area cultivated with Brachiaria grass in Londrina, state of Paraná, Brazil (23°34ʹ311ʹʹS, 050°58ʹ298ʹʹW), using the insect-baiting technique with Galleria mellonella larvae as hosts. The nematode was identified through morphometric studies and molecular analyses based on amplification of the rDNA ITS region. Although we identified certain morphometric differences compared with the original description, the molecular data indicated that the ITS sequence obtained for the UEL 08 isolate is identical to the reference sequence of H. amazonensis (DQ665222) and presented 100% similarity. Thus, the findings of our morphological and molecular studies confirmed that the isolated nematode is H. amazonensis, which is the first time this species has been registered in Paraná. Study of the biological characteristics of H. amazonensis (UEL 08) revealed that the isolate has two distinct life cycles – one short (216 h) and the other long (288 h) – and produces two generations in both cycles. We observed that H. amazonensis (UEL 8) was pathogenic and virulent to the three evaluated hosts, although with different virulence against these hosts. The larvae of G. mellonella and Alphitobius diaperinus were more susceptible than adult Dichelops (Diacereus) melacanthus, with 100%, 85%, and 46% mortality, respectively. Furthermore, an in vivo production assay revealed a mean daily yield of 3.4 × 103 infective juveniles/g host larvae.

Formulation of entomopathogens refers to the mixing of various inert ingredients, like clays and mineral oils, with the active ingredients which are the entomopathogens. Successful formulation enhances the survival of the entomopathogen and eases their transportation, storage, preparation and application. The aim of this study was to develop a formulation to maintain the longevity and pathogenicity of the mass-produced conidia of local Metarhizium pinghaense and M. robertsii, for above-ground future commercial field application against Pseudococcus viburni. The objectives were to develop a cost-effective protocol for formulation of infective propagules and to test their effectiveness under laboratory conditions. The conidia of both isolates were formulated using four different oils (liquid paraffin, coconut, canola and olive oils) as liquid carriers, and diatomaceous earth as a mineral carrier. Conidial viability and pathogenicity were assessed over a period of eight weeks. Conidia formulated in oil carriers maintained a high conidial viability and survival rate of > 95% over a period of eight weeks for both isolates, relative to when formulated in mineral carriers, or when stored as dry conidial powder. The conidia in all the oil formulations were also observed to induce high mortality, ranging between 60% and 90% for M. pinghaense, and between 70% and 90% for M. robertsii, when used against P. viburni. The ability of conidia of both isolates to maintain viability and pathogenicity, following storage in the oil formulations, increased the likelihood of the local isolates being successfully integrated as biological control agents for management of P. viburni under field conditions.

Resumen El género Beauveria comprende un grupo cosmopolita teleomórfico y anamórfico de hongos entomopatógenos. El objetivo de esta investigación fue caracterizar un nuevo aislado de Beauveria sp. en comparación con la cepa comercial cubana LBB-1. El aislamiento se obtuvo de diez individuos de Atta insularis y diez adultos del barrenador del café (Hypotenemus hampei), respectivamente. Se cultivó en medio sólido SDAY enriquecido con 1% de extracto de levadura. Se determinó la eficacia biológica del aislado sobre C. formicarius. El ADN se aisló usando el método CTAB y se amplificó la región de espaciador interno transcrito usando los cebadores ITS1 e ITS4. La secuencia consenso obtenida se depositó en GenBank (MK246940) y se utilizó para análisis filogenético. El aislado de Beauveria sp. mostró diferencias en sus características morfológicas; sin embargo, mostró una baja variabilidad genética con respecto a los aislamientos mundiales de B. bassiana. En cuanto a su eficacia biológica, el aislado demostró ser virulento sobre los adultos fitófagos, alcanzando el 50% de mortalidad. Se identificaron cuatro clados y la topología politómica recuperada para el clado 1 indica una alta similitud genética (99-96%) con Beauveria bassiana de varios países. La caracterización morfológica y molecular permitió identificar con precisión la especie del nuevo aislado.

Summary The genus Beauveria comprises a cosmopolitan teleomorphic and anamorphic group of entomopathogenic fungi. The aim of this study was to characterize a new isolate of Beauveria sp. and compare it to the LBB-1 commercial Cuban strain. The isolate was collected from ten individuals of the ant Atta insularis and ten adults of the coffee borer (Hypotenemus hampei). It was cultivated on solid SDAY medium enriched with 1% yeast extract. The biological effectiveness of the isolate was determined on C. formicarius. DNA was isolated using the CTAB method and the internal transcribed spacer region was amplified using the primers ITS1 and ITS 4. The consensus sequence obtained was deposited in GenBank (MK246940) and used for phylogenetic analysis. The isolate of Beauveria sp. showed differences in morphological characteristics; however, it showed low genetic variability regarding worldwide isolates of B. bassiana. Concerning its biological effectiveness, the isolate demonstrated to be virulent on the phytophagous adults, producing 50% mortality. Four clades were identified and the polytomous topology recovered for clade 1 indicates high genetic similarity (99-96%) with Beauveria bassiana from several countries. The morphological and molecular characterization made it possible to accurately identify the new isolate as Beauveria bassiana.

ABSTRACT Lime sulfur is one of the few products indicated to control Brevipalpus yothersi in Brazilian organic citrus orchards. Other strategies, such as the use of entomopathogenic fungi should be evaluated, and Lecanicillium muscarium is one of the basic choices for pest management. Knowledge of the interactions between lime sulfur and this entomopathogen is critical for developing control strategies. With this goal, it was conducted the toxicological characterization of lime sulfur to B. yothersi and the compatibility evaluation with L. muscarium. Finally, the effects of L. muscarium and lime sulfur mixtures on B. yothersi control were evaluated. Product evaluation for B. yothersi was done through direct and residual contact bioassay, and different concentrations of lime sulfur mixed in potato dextrose agar culture medium were used to evaluate compatibility with L. muscarium. Lime sulfur was effective against adults of B. yothersi and caused eggs unviability of up to 71.0%, at a dose of 80 L per 2,000 L of H2O. The lethal concentration (LC50 and LC99) of lime sulfur estimated for mite adults were 246.62 and 858.5 μg of sulfur per mL of H2O (ppm a.i.). Lime sulfur concentrations of 180 to 560 ppm a.i. showed promise for use in combination with L. muscarium. However, concentrations of 1,000 and 5,600 ppm significantly reduced colony size and the number of spores/colony. The mixture of 100 and 180 ppm a.i. of lime sulfur with L. muscarium (108 conidia·mL–1) was not able to reduce the lethal time of entomopathogen on B. yothersi.

Resumo Bacillus thuringiensis é o entomopatógeno mais utilizado no controle do Aedes aegypti, vetor de diferentes agentes etiológicos que causam infecções graves em humanos. Diversos estudos têm como objetivo isolar cepas dessa bactéria de diferentes ambientes, com a perspectiva de selecionar isolados com atividade larvicida para mosquitos. Visando a ação inseticida de B. thuringiensis, o presente estudo teve como objetivo prospectar B. thuringiensis de solos de restinga e mangue do estado do Maranhão, Brasil, com potencial tóxico para uso no controle biológico de Ae. aegypti. Bioensaios foram realizados para determinar a atividade entomopatogênica do bacilo contra Ae. aegypti e as concentrações letais (CL50 e CL90) foram estimadas após os testes. As técnicas de Reação em Cadeia da Polimerase e SDS-PAGE foram realizadas para verificar o conteúdo de genes e proteínas dos isolados, respectivamente. Os solos dos ecossistemas de mangue e restinga apresentaram potencial para obtenção de B. thuringiensis. O isolado BtMA-750, obtido a partir da amostra de solo da restinga, foi interessantemente distinguido por sua alta toxicidade para Ae. aegypti. Este isolado, além de apresentar proteínas com massa molecular semelhante às toxinas Cry e Cyt, apresentou também diversos genes díptero-específicos cry e cyt, demonstrando que tem alto potencial para ser usado no controle biológico de Ae. aegypti.

Abstract Bacillus thuringiensis is the most commonly used entomopathogen in the control of Aedes aegypti, which is a vector for different etiological agents that cause serious infections in humans. Several studies aim to isolate strains of this bacterium from different environments, with the perspective of selecting isolates with larvicidal activity for mosquitoes. Aiming at the insecticidal action of B. thuringiensis, the present study aimed to prospect B. thuringiensis of restinga and mangrove soils from the state of Maranhão, Brazil, with toxic potential for use in the biological control of Ae. aegypti. Bioassays were performed to determine the entomopathogenic activity of the bacilli against Ae. aegypti and lethal concentrations (LC50 and CL90) were estimated after the tests. Polymerase Chain Reaction and SDS-PAGE techniques were performed to verify the gene and protein content of the isolates, respectively. The soil of the mangrove and restinga ecosystems showed potential for obtaining B. thuringiensis. This isolate, in addition to having proteins with molecular mass similar to the toxins Cry and Cyt, also presented several diptera-specific genes cry and cyt, demonstrating that it has high potential to be used in the biological control of Ae. aegypti.

Resumo O controle biológico é uma tendência mundial e tem se destacado como uma estratégia eficaz e compatível para uso em programas de Manejo Integrado de Pragas. No Brasil, o controle desses organismos, envolvendo agentes biológicos, tem sido utilizado em diversos sistemas agrícolas, inclusive, na floricultura. Neste artigo, o controle biológico é abordado sob o ponto de vista natural, por meio da interação entre os organismos na natureza, e aplicado, como estratégia para o manejo de pragas. O conhecimento sobre a dependência e reciprocidade existentes entre as populações na natureza é fundamental para a conscientização de que esse método de controle consiste na exploração de um serviço ecológico natural. Tendo-se em vista que, para o controle biológico de uma praga, a densidade populacional dos inimigos naturais deve ser aumentada, relatamos sobre as duas principais formas para se atingir esse incremento: o controle biológico conservativo e controle biológico aumentativo. A primeira, por meio da modificação do ambiente de cultivo de modo a oferecer uma estrutura vegetacional que favoreça a manutenção e/ou atração dos inimigos naturais, e a segunda, por meio da criação massal de entomófagos e entomopatógenos com características que possam garantir sua função como agentes de controle por ocasião da sua liberação nos cultivos. Entre tais agentes, enfatizamos aqueles que são produzidos e comercializados no Brasil para aplicação em plantas ornamentais. Por fim, relatamos sobre as tecnologias que têm sido mais recentemente empregadas para incrementar o uso adequado e a eficiência desses agentes biocontroladores.

Abstract Biological control is a worldwide trend and has been prominent as an effective and compatible strategy for use in Integrated Pest Management programs. In Brazil, the control of these organisms using biological agents has been used in several agricultural systems, including floriculture. This paper approaches biological control from the perspective of the interaction between organisms in nature, applied as a pest management strategy. Knowledge about the dependence and reciprocity among populations in a natural environment is fundamental to understanding that this control method is nothing more than an exploiting of a natural ecological service. Considering that, for the biological control of a pest we must increase the population density of natural enemies, we report on the two main ways to achieve this increase: conservative biological control and augmentative biological control. The first is done by modifying the environment’s vegetation structure to favor natural enemies’ maintenance or their attraction to the crops. The second is done by mass rearing entomophagous and entomopathogen species with attributes that can ensure their function as control agents when released in crops. Among such agents, we emphasize those produced and marketed in Brazil for application in ornamental plants. Finally, we report on the technologies most recently employed to increase the appropriate use and the efficiency of these biological control agents.

Resumen Introducción: Los hongos entomopatógenos se han utilizado como agentes de control biológico de plagas de insectos y han demostrado una capacidad prometedora para colonizar una amplia gama de especies de plantas como endófitos. Objetivo: Los objetivos de este estudio fueron: evaluar las técnicas de inoculación más efectivas para B. bassiana en plantas de pimiento; evaluar la presencia de endófitos naturales y su interacción con B. bassiana y evaluar el efecto de B. bassiana sobre la preferencia de alimentación de la hoja de pimiento y el crecimiento de la plaga de áfidos M. persicae. Métodos: Se evaluaron tres técnicas de inoculación: aspersión foliar, inmersión de semillas e inmersión de raíces. Las plantas de pimiento se inocularon con una suspensión de conidias ajustada a 1x108 conidias/ml. La presencia de B. bassiana en las plántulas se evaluó a los 7, 15 y 28 días después de la inoculación. Resultados: Se encontraron diferencias significativas entre las técnicas de inoculación. La aspersión foliar mostró la frecuencia más alta. Los endófitos naturales presentaron frecuencias más altas en los tratamientos en los que B. bassiana tuvo una frecuencia baja, esto podría indicar la presencia de actividad antifúngica que interfiere con el establecimiento del entomopatógeno en los tejidos de pimiento. No se encontraron diferencias significativas en la mortalidad, reproducción y preferencia alimentaria de M. persicae. Conclusiones: Se demostró que la cepa B. bassiana LPSC 1067 se estableció efectivamente como endófito en los tejidos de pimiento. La técnica de inoculación más efectiva fue la aspersión foliar. No se encontraron diferencias significativas con respecto a la mortalidad, reproducción y preferencia alimentaria de M. persicae. Sin embargo, la inoculación con B. bassiana mostró una tendencia a aumentar la mortalidad de los adultos y a disminuir la descendencia (ninfas). Se necesitan estudios futuros que evalúen los efectos indirectos de los entomopatógenos endofíticos tanto en plantas como en insectos herbívoros para explorar más a fondo su posible uso en el manejo de insectos plaga.

Abstract Introduction: Entomopathogenic fungi have been used as biological control agents of insect pests and have demonstrated a promising ability to colonize as endophytes a wide range of plant species. Objective: The aims of this study were to evaluate, in pepper plants, the most effective Beauveria bassiana inoculation techniques. In order to evaluate the presence of natural endophytes and its interaction with B. bassiana. Also to assess the effect of B. bassiana on the aphid Myzus persicae pepper leaf feeding preference and growth. Methods: Three inoculation techniques were evaluated: leaf spraying, seed immersion and root dipping. The pepper plants were inoculated with a conidia suspension adjusted to 1×108 conidia/ml. The presence of B. bassiana in the seedlings was evaluated at 7, 15 and 28 days after inoculation. Results: We found significant statistical differences between inoculation techniques. Foliar spraying showed the highest B. bassiana colonizationfrequency. The natural endophytes presented higher presence in the treatments in which B. bassiana had a low presence. This might indicate the occurrence of antifungal activity that interferes with the establishment of the entomopathogen in pepper tissues. We did not find statistical differences between inoculated and control pepper leaves with regard to the mortality and reproduction of the aphid pest M. persicae. However, the inoculation with B. bassiana showed a tendency to increase the mortality of adults and to decrease the offspring (nymphs). We did not find significant differences in the feeding preference between inoculated and control leaves, although there was a higher number of individuals on control leaves compared to those inoculated. Conclusions: The strain B. bassiana LPSC 1067 was established effectively as an endophyte in the pepper tissues. The most effective inoculation technique was leaf spraying. We did not find differences of statistical significance with regard to the mortality and reproduction of the aphid pest M. persicae. However, the inoculation with B. bassiana showed a tendency to increase the mortality of adults and to decrease the offspring (nymphs). Future studies evaluating the indirect effects of endophytic entomopathogens on both plants and herbivorous insects are needed to further explore their potential use in the management of insect pests.

Abstract Leptolegnia chapmanii is pathogenic to mosquito larvae. The compatibility betweenL. chapmanii and two insect growth regulators (IGR), diflubenzuron and neem oil, was evaluated. L. chapmanii was grown on culture media containing different concentrations of eachIGR. The mycelial growth was significantly reduced with the highest concentrations of IGR(F6,98= 268, p < 0.05). However, the production of zoospores (F6,56= 0.93, p > 0.05) and the lar-val mortality of Aedes aegypti (F6,56= 0.95, p > 0.05) were not significantly different amongtreatments. Furthermore, the percentage of adult emergence in the presence of different con-centrations of diflubenzuron or a neem formulation was determined, and the pathogenic activityof zoospores was evaluated at the concentrations that inhibit the emergence at 30, 50 and 90%.The pathogenicity of zoospores was not significantly different among treatments (F6,14= 0.54,p > 0.05), and the larval mortalities were above 90% in all cases.

Resumen Leptolegnia chapmanii es un patógeno de larvas de mosquitos. Se evaluó la com-patibilidad entre L. chapmanii y dos reguladores del crecimiento de insectos (IGR, por sus siglasen inglés), diflubenzurón y aceite de neem. L. chapmanii creció en medios de cultivo con diferentes concentraciones de estos IGR. El crecimiento micelial fue menor con las concentraciones más altas (F6,98= 268; p < 0,05). No hubo diferencias significativas en la producción de zoosporas (F6,56= 0,93; p > 0,05) ni en la mortalidad de las larvas de Aedes aegypti (F6,56= 0,95; p > 0,05).Se determinó el porcentaje de emergencia de adultos en presencia de diferentes concentra-ciones de diflubenzurón o de un formulado a base del aceite de neem. También se evaluó lapatogenicidad de las zoosporas a las concentraciones de los IGR que inhibieron la emergenciadel 30, 50 y 90% de adultos. No hubo diferencias significativas en la actividad de las zoosporasal comparar los tratamientos (F6,14= 054; p > 0,05). La mortalidad de las larvas fue superior al90%.

Resumen Monalonion velezangeli (Hemiptera: Miridae) es una plaga polífaga que en Colombia ataca ornamentales, arvenses, maderables y frutales incluyendo café. Se evaluó la patogenicidad de Beauveria bassiana cepa B a 1 x 107 conidias/ml por inmersión sobre M. velezangeli. La cepa B causó una mortalidad de 84 % sobre el insecto. Se infestaron plantas de Cissus verticillata con M. velezangeli y se aplicó el hongo a una concentración de 4 x 1010 conidias/L. La mortalidad promedio de los insectos, bajo condiciones de laboratorio en el tratamiento con el hongo fue de 84 %. En condiciones de campo, en el departamento del Huila, en un lote de café infestado con M. velezangeli, se aplicaron los tratamientos: cepa B a una concentración de 4 x 1010 conidias/L, insecticida Clorpirifos a 4 cc/L y agua. En un primer ensayo se escogieron grupos de nueve árboles (20 repeticiones). Tanto el insecticida como el hongo causaron mortalidad en los insectos con disminución entre 80 y 90 % de los porcentajes de daños nuevos en los árboles, mientras que en los testigos la disminución del daño fue de 40 %. En un segundo ensayo, se seleccionaron parcelas de 450 árboles en donde se aplicaron los mismos tratamientos. El porcentaje de árboles afectados en el testigo varió entre 19 y 11 %. El lote con aplicación de insecticida, inició con una infestación del 29 % y, al final del experimento, la infestación estaba cercana al 20 % y el mayor daño fue observado en este lote. En el lote donde se aplicó B. bassiana, la infestación inicial fue de 30 % y al final del experimento mostró el menor nivel de infestación, cercano al 5 %. La cepa B de B. bassiana representa una alternativa viable para el control de M. velezangeli en plantaciones de café.

Abstract Monalonion velezangeli (Hemiptera: Miridae) is a polyphagous pest found in Colombia which attacks ornamentals, weeds, timber and fruit trees including coffee. The pathogenicity of Beauveria bassiana strain B at 1 x 107 conidia/ml was evaluated on Monalonion velezangeli by immersion. The fungus caused 84 % mortality on the insect. Cissus verticillata plants were infested with M. velezangeli and applications of the fungus at 4 x 1010 conidium/L were done. Average insects mortality treated with the fungus was 84 %. Under field conditions, in the department of Huila, in a coffee plot infested with M. velezangeli the following treatment were applied: strain B at 4 x 1010 conidium/L, insecticide Chlorpyrifos at 4 cc/L and water. In a first assay, groups of nine trees (20 repetitions) were chosen. Both the insecticides and the fungus caused insect mortality with an 80 to 90 % decrease in the percentage of trees with new damage. Meanwhile the decrease in the control was 40 %. In a second assay, three plots of 450 trees were selected and the same treatments were applied. Percentage of trees affected in the control ranged between 19 and 11 %. The plot in which the insecticide was applied, started with an infestation of 29 %, and at the end of the assay the infestation was 20 % and the highest damage was observed in this plot. In the plot where B. bassiana was applied the initial infestation was 30 % and at the end of the assay, this plot showed the lowest infestation level near to 5 %. The B. bassiana strain B becomes a feasible alternative for M. velezangeli pest control on coffee plantations.

ABSTRACT: The citrus scale insect Praelongorthezia praelonga (Douglas), a major pest of citrus and other economically important crops, has only two commonly documented natural enemies: an entomopathogenic strain of the fungus Colletotrichum nymphaeae (Pass.) Aa and several parasitoids. The entomopathogenic strain of C. nymphaeae, formerly recognized under the synonym C. gloeosporioides f. sp. ortheziidae, is under development for commercial application as a biological control agent in citrus in Brazil-the top exporter of citrus globally. The synonomy of C. gloeosporioides f. sp. ortheziidae with C. nymphaeae remains based on limited DNA sequence data and without morphological study. To qualify for future approval as a biological control agent by federal agencies in Brazil and the European Union, the circumscription of a microorganism must be explicit and without ambiguities. Herein, through morphological study and phylogenetic analysis of five DNA regions we clarify the circumscription and affinity of entomopathogenic C. nymphaeae and describe it as a new variety.

Abstract Beauveria bassiana is an entomopathogenic fungus that produces secondary metabolites as virulence factors during the pathogenic process in insects. In this study, eleven strains of B. bassiana were analyzed to determine the presence of the bbBeas and bbBsls genes that produce beauvericin and bassianolide non-ribosomal cyclooligomeric depsypeptides, respectively. The genes were characterized by PCR and DNA sequencing. The presence of beauvericin and bassianolide from mycelial extracts were determined by mass spectrometry. The results indicated that with the exception of strain 2857, they all possess both genes and synthesized bassianolide. Beauvericin was not produced in any case. Mortality caused by conidia in Spodoptera exigua and S. frugiperda neonate larvae was variable. Strains 1512, 2253 and 2857 caused significantly higher mortalities (44 % mortality on average). The bassianolide extract injected (1 mg/mL) caused 26 % mortality in S. exigua. The lack of beauvericin production, with the exception of strain 1149, may be due to the requirement of other culture conditions that stimulate the expression of the genes. So far, no correlation is observed between the pathogenic activity of the fungus, and the presence or absence of these secondary metabolites.

Resumen Beauveria bassiana es un hongo entomopatógeno que produce metabolitos secundarios como factores de virulencia durante el proceso patogénico en insectos. En este estudio se analizaron once cepas de B. bassiana para determinar la presencia de los genes bbBeas y bbBsls productores de los depsipéptidos ciclooligoméricos no ribosomales beauvericina y bassianolida, respectivamente. Los genes fueron caracterizados por PCR y secuenciación. La presencia de beauvericina y bassianolida en extractos de micelio se determinó por espectrometría de masas. Los resultados indicaron que con excepción de la cepa 2857, todas poseen los dos genes y sintetizan bassianolida. En ningún caso se produjo la beauvericina. La mortalidad por conidias en larvas de Spodoptera exigua y S. frugiperda fue variable, pero las cepas 1512, 2253 y 2857 causaron las mayores mortalidades (44 % en promedio). El extracto con bassianolida inyectado (1 mg/mL) causó 26 % de mortalidad en larvas de S. exigua. La falta de producción de beauvericina, con excepción de la cepa 1149, puede deberse al requerimiento de otro tipo de condiciones de cultivo que estimulen la expresión de los genes. Hasta ahora no se observa correlación entre la actividad patogénica del hongo y la presencia o ausencia de estos metabolitos secundarios.

ABSTRACT Context: Sunflower is the fourth source of vegetable oil in the world. The sunflower moth is its main pest in America. The control of this species is extremely difficult and, generally, the solutions proposed for its control involve the use of insecticides. Objective: To formulate an ecologically sustainable strategy for the control of the sunflower moth. Methods: The insect-host interaction and the damage caused were characterized. The best biological control agents were selected in terms of efficacy. Results: None of the tested cultivars was resistant to Homoeosoma electellum. The perfect timing to protect the sunflower against H. electellum might be estimated between 65 % of the life cycle of the cultivars until reaching the physiological maturity and 84 % until reaching the same phenological state. The application of entomopathogenic bio-preparations significantly reduces the affectation. Conclusions: Treatment with entomopathogenic bio-preparations during the delimited critical interval allows controlling the damage caused to sunflower.

RESUMEN Contexto: El girasol ocupa el cuarto lugar entre las fuentes de aceite vegetal. Su principal plaga en América es la polilla del girasol. Esta especie es extremadamente difícil de controlar y las soluciones propuestas implican el uso de insecticidas. Objetivo: Formular una estrategia ecológicamente sostenible para el control de la polilla del girasol. Métodos: Se caracterizó la interacción insecto-hospedante, los daños ocasionados y se seleccionaron los mejores agentes de control biológico en términos de eficacia. Resultados: Ninguno de los cultivares probados resultó tolerante a Homoeosoma electellum. Un intervalo seguro para proteger al girasol contra H. electellum puede estimarse entre el 65 % de la duración del ciclo de los cultivares hasta alcanzar madurez fisiológica y el 84 % hasta alcanzar el mismo fenoestado. La aplicación de biopreparados entomopatógenos Heterorhabditis indica (Cepa P2M), Beauveria bassiana (Cepa LbB-32) y Metarhizium anisopliae (Cepa LbM-11) reduce significativamente la afectación. Conclusión: El tratamiento con biopreparados entomopatógenos durante el intervalo crítico indicado anteriormente permite controlar los daños producidos por la polilla del girasol.

Resumen Introducción. En Latinoamérica, el picudo (Rhynchophorus palmarum L.) es plaga de importancia en palma de aceite y cocotero, porque se asocia con la transmisión de anillo rojo (AR) y la pudrición del cogollo (PC), enfermedades que causan altos costos de manejo y mortalidad de plantaciones; los entomopatógenos Metarhizium anisopliae y Beauveria bassiana (Hypocreales: Clavicipitaceae) representan una alternativa de control microbiológico. Objetivo. El objetivo de este trabajo fue determinar la patogenicidad de cepas promisorias de M. anisopliae y B. bassiana sobre adultos de R. palmarum, y evaluar la diseminación de las mejores cepas desde adultos machos del picudo inoculados hacia hembras sin inocular. Materiales y métodos. El estudio se desarrolló en dos fases en el Centro de Investigación La Libertad de la Corporación Colombiana de Investigación Agropecuaria (AGROSAVIA), Departamento del Meta, Colombia, mediante arreglos factoriales en diseños completamente al azar; en la primera, se evaluaron cuatro cepas de M. anisopliae, una de B. bassiana y un producto comercial a base de M. anisopliae; en la segunda, para determinar la autodiseminación del entomopatógeno, se evaluaron las dos cepas de mayor eficacia del primer experimento (CPMa1104 y Mt008), mediante dos métodos de aplicación a machos (inmersión y polvo). Resultados. La cepa CPMa1104 mostró la mayor eficacia (1x104 y 1x108 conidias ml-1), seguida de la Mt008 (1x108 conidias ml-1), con mortalidades de 78,6 % y 67,9 %, respectivamente. Se observó capacidad de diseminación por contacto de M. anisopliae cepas CPMa1104 y Mt008, desde adultos machos inoculados hacia hembras sin inocular. La mortalidad de las hembras alcanzó 100 % cuando los machos fueron inoculados con la cepa CPMa1104 aplicada en polvo. Conclusión. Se comprobó la diseminación de las cepas CPMa1104 y Mt008 de M. anisopliae por machos inoculados hacia hembras sin inocular, lo cual convierte el uso de este entomopatógeno en una alternativa promisoria de control.

Abstract Introduction. In Latin America, the weevil (Rhynchophorus palmarum L.) is an important pest of oil palm and coconut palm because it is associated with the transmission of red ring (RRD) and bud rot (BR); diseases that cause high management costs and mortality of plantations; the entomopathogens Metarhizium anisopliae and Beauveria bassiana (Hypocreales: Clavicipitaceae) represent an microbiological control alternative. Objective. The objective of this work was to determine the pathogenicity of promising strains of M. anisopliae and B. bassiana on adults of R. palmarum and to evaluate the spread of the best strains, from inoculated weevil male adults to not-inoculated females. Materials and methods. The study was carried out in two phases in the La Libertad Research Center of the Corporación Colombiana de Investigación Agropecuaria (AGROSAVIA), Department of Meta, Colombia through factorial arrangements in completely random designs; in the first one, four strains of M. anisopliae, one of B. bassiana, and one commercial product based on M. anisopliae were evaluated; in the second one, to determine the self-dissemination of the entomopathogen, the two highest effectiveness strains from the first experiment (CPMa1104 and Mt008) were evaluated by two methods of application to males (immersion and powder). Results. The CPMa1104 strain showed the highest efficacy (1x104 and 1x108 conidia ml-1), followed by Mt008 (1x108 conidia ml-1) with mortalities of 78.6 % and 67.9 %, respectively. Contact dissemination capacity of M. anisopliae strains CPMa1104 and Mt008 was observed from inoculated male adults to non-inoculate females. Female mortality reached 100 % when the males were inoculated with powdered CPMa1104. Conclusion. The dissemination of CPMa1104 and Mt008 strains of M. anisopliae by inoculated males to non-inoculated females was tested, which makes the use of this entomopathogen a promising control alternative.