Aspectos biológicos de Orius insidiosus (Say, 1832) (Hemiptera: Anthocoridae) alimentado com larvas de Plutella xylostella (Linnaeus, 1758) (Lepidoptera: Plutellidae) criadas em brassicáceas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Behavior of Orius insidiosus preying on Aphis gossypii reared on transgenic and conventional cotton varieties

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Record of two species of Orius Wolff (Hemiptera, Anthocoridae) in Brazil

The genus Orius Wolff, 1811 comprises predatory species, with approximately 70 known species. Informations about the genus in Brazil are scarce. Therefore, the aim of this investigation was to identify Orius species present in four localities in the southeastern Brazil. Samples were taken from several plants, and the material screened in laboratory. The genitalia of both sexes were studied and illustrated. Two species were identified, Orius insidiosus (Say, 1832) and Orius thyestes Herring, 1966. O. insidiosus, the most common species, was collected in all of the localities sampled [Lavras (MG), Holambra, Pindorama and Campinas (SP)]. O. thyestes, registered for the first time in Brazil, occurred only in Lavras (MG) and Pindorama (SP). Some morphologic aspects of these two species are also presented.

Percevejos predadores (Orius spp.) (Hemiptera: Anthocoridade) e tripes (Thysanoptera): interação no mesmo habitat?

O gênero Orius Wolff é composto por espécies de percevejos predadores, principalmente de tripes, encontradas em muitos ecossistemas naturais e manejados. Entretanto, as interações que podem ocorrer entre esses predadores e suas presas no mesmo "habitat" não são bem conhecidas entre as espécies das regiões tropicais. Este estudo teve como objetivo registrar a interação de espécies de Orius e de tripes coletadas na mesma planta, ou seja, presentes no mesmo "habitat". As coletas foram feitas em várias plantas cultivadas, no campo e em casas de vegetação e em plantas invasoras. A forma e o grau de associação entre as espécies foram determinados, utilizando-se o coeficiente de Spearman (R), calculado com dados de presença/ausência das espécies (Orius e tripes) no mesmo "habitat". Orius insidiosus (Say) foi encontrado associado positivamente aos tripes Frankliniella sp., Neohydatothrips sp. e Haplothrips gowdeyi (Franklin) e negativamente associado a Frankliniella schultzei (Trybom) e Caliothrips phaseoli (Hood). Orius thyestes Herring e Orius sp1 ocorreram simultaneamente em 10 espécies de tripes sem, contudo, apresentar associação significativa, enquanto Orius perpunctatus (Reuter) esteve associado positivamente às espécies Frankliniella sp. e Neohydatothrips sp. e, negativamente, a Frankliniella gemina (Moulton).

Soil organisms associated to the weed suppressant Crotalaria juncea (fabaceae) and its importance as a refuge for natural enemies

Soil organisms play an important role in organic crops of Crotalaria juncea (Fabaceae) and are associated with the natural conservation of the environment. The present study was aimed to investigate the population of soil organisms in the organic culture of C. juncea, as well as its importance as a refuge for natural enemies. Dalbulus maidis (Hemiptera: Cicadellidae), Diabrotica sp. (Coleoptera: Chrysomelidae), Doru luteipes (Dermaptera: Forficulidae), Gryllus assimilis (Orthoptera: Gryllidae), Lagria villosa (Coleoptera: Lagriidae), Melanotus sp. (Coleoptera: Elateridae), Meloidogyne incognita (Tylenchida: Heteroderidae), Nephila clavipes (Araneae: Nephilidae), Orius insidiosus (Hemiptera: Anthocoridae), Pheidole sp. (Hymenoptera: Myrmicidae), Phyllophaga sp. (Coleoptera: Scarabeidae), Procornitermes sp. (Isoptera: Termitidae), Solenopsis sp. (Hymenoptera: Formicidae), and Utetheisa ornatrix (Lepidoptera: Arctiidae) were identified in C. juncea. The organisms that were found during a 3-month period in 144 trenches in C. juncea were pest species (84.47%) and natural enemies (15.53%) as well. Natural enemies had an average of 11.89 individuals per 1.08 m³ of soil cultivated with C. juncea. The abundance of organisms in the pod stage (5.49%) of C. juncea was lower than that in the vegetative (83.50%) and flowering (11.01%) stages. Crotalaria juncea plants can be used as part of a crop system for Integrated Pest Management.

PREDATION OF COTTON LEAFWORM 1ST INSTAR LARVAE, ALABAMA-ARGILLACEA [LEP, NOCTUIDAE]

Natural predation first instar larvae of the cotton leafworm (CLW) A. argillacea was studied in cotton fields in Jaboticabal, São Paulo State, Brazil, during 1986. The presence of naturally occurring arthropod predators showed a first instar larvae predation rate of 78.6 and 88.9% after 24 h and 48 h of exposure, respectively. A predator prey ratio of 1 : 1 (1 CLW key predator per 1 prey/plant) maintained a level of no more than 1 CLW small larvae per plant. The most evident arthropod predators in the studied fields were: beetles (Coleoptera: Coccinellidae), ants Pheidole sp. and Conomyrma sp.; Dermaptera Doru lineare (Eschs); Hemiptera Geocoris sp., and Orius insidiosus Say; and the spiders Theridion volubile, Chrysso pulcherrima, Misumenops sp., Chiracanthium sp., and Oxyopes salticus Hentz.

Fatores naturais e Telenomus remus Nixon (Hymenoptera: Scelionidae) na mortalidade de Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) em milho

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ação de Bacillus thuringiensis Berliner nas características biológicas de outrs inimigos naturais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ecologia química no controle biológico de fitófagos e respectivos predadores e parasitoides

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Guia para a identificação e monitoramento de pragas e seus inimigos naturais em morangueiro.

2015

Influência da temperatura na reprodução e longevidade do predador Orius thyestes Herring (Hemiptera, Anthocoridae)

O desenvolvimento e a fecundidade das espécies de Orius são bastante influenciados por uma série de fatores, como as condições ambientais, e em particular, pela temperatura. Esse trabalho teve como objetivo avaliar a reprodução e a longevidade de Orius thyestes Herring 1966 em diferentes temperaturas, tendo como alimento, ovos de Anagasta kuehniella (Zeller, 1879). O experimento foi conduzido em câmaras climatizadas com temperaturas de 16, 19, 22, 25, 28, 31 ± 1ºC, UR de 70±10% e fotofase de 12horas. Efeito deletério da temperatura em O. thyestes foi obtido a 16ºC, na qual apenas 40% das ninfas atingiram a fase adulta, e destes apenas 19% não apresentaram deformações morfológicas. O maior período de pré-oviposição foi observado a 19ºC (17,8 dias). Os maiores valores para a fecundidade média total foram registrados a 25 e 28ºC, com 109,2 e 128,2 ovos/fêmea, respectivamente, e o menor a 19ºC, com 22,8 ovos/fêmea. A 22 e 31ºC as fêmeas viveram mais que os machos, sendo que a 19ºC a longevidade foi maior, independente do sexo. As baixas temperaturas influenciaram a reprodução e longevidade de O. thyestes sugerindo que esta espécie poderá ter melhor performance reprodutiva em temperaturas mais elevadas, como aquelas de regiões tropicais e ou subtropicais.

Selectivity of diacylhydrazine insecticides to the predatory bug Orius laevigatus: In vivo and modeling/docking experiments

BACKGROUND: Knowledge of pesticide selectivity to natural enemies is necessary for a successful implementation of biological and chemical control methods in integrated pest management (IPM) programs. Diacylhydrazine (DAH)-based ecdysone agonists also known as molting-accelerating compounds (MACs) are considered a selective group of insecticides, and their compatibility with predatory Heteroptera, which are used as biological control agents, is known. However, their molecular mode of action has not been explored in beneficial insects such as Orius laevigatus (Fieber) (Hemiptera: Anthocoridae). RESULTS: In this project in vivo toxicity assays demonstrated that the DAH-based RH-5849, tebufenozide and methoxyfenozide have no toxic effect against O. laevigatus. The ligand-binding domain (LBD) of the ecdysone receptor (EcR) of O. laevigatus was sequenced and a homology protein model was constructed which confirmed a cavity structure with 12 ?-helixes, harboring the natural insect molting hormone 20-hydroxyecdysone. However, docking studies showed that a steric clash occurred for the DAH-based insecticides due to a restricted extent of the ligand-binding cavity of the EcR of O. laevigatus. CONCLUSIONS: The insect toxicity assays demonstrated that MACs are selective for O. laevigatus. The modeling/docking experiments are indications that these pesticides do not bind with the LBD-EcR of O. laevigatus and support that they show no biological effects in the predatory bug. These data help in explaining the compatible use of MACs together with predatory bugs in IPM programs. Keywords: Orius laevigatus, selectivity, diacylhydrazine insecticides, ecdysone receptor, homology modelling, docking studies.

Compatibilidad de Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) y Nesidiocoris tenuis (Reuter)(Hemiptera: Miridae), depredadores importantes en cultivos hortícolas protegidos, con nuevas barreras físicas selectivas y modernos plaguicidas

La nueva legislación en materia fitosanitaria se dirige hacia una Gestión Integrada de Plagas (GIP). Estos programas dan preferencia a aquellos métodos más respetuosos y sostenibles con el medio ambiente, siendo piezas claves en ellos el control biológico, el físico y otros de carácter no químico. Sin embargo, el uso de insecticidas selectivos es a veces necesario para el adecuado manejo de plagas en cultivos hortícolas. Por ello, el objetivo general de este estudio es aportar conocimientos para mejorar el control de plagas en cultivos hortícolas, mediante la integración de tres estrategias de lucha: biológica, física y química. Una parte de este trabajo ha consistido en el estudio de los posibles efectos que mallas tratadas con insecticida (bifentrin) pudieran provocar mediante diferentes ensayos de laboratorio, invernadero y campo, en los enemigos naturales Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) (depredador de trips), Nesidiocoris tenuis (Reuter) (Hemiptera: Miridae) (depredador de mosca blanca y Tuta absoluta (Meirick) (Lepidoptera: Gelechiidae)), y otros agentes de biocontrol comúnmente usados en cultivos hortícolas protegidos. Este tipo de mallas se han empleado con éxito en entomología médica para controlar mosquitos vectores de la malaria, y actualmente se está trabajando en su desarrollo para uso agrícola como método de exclusión, y método directo de control de plagas. En los ensayos realizados en laboratorio, O. laevigatus y N. tenuis no fueron capaces de detectar la presencia de bifentrin en el ensayo de preferencia. Además, no se produjo mortalidad a corto plazo (72 horas) en ambos chinches depredadores. Por el contrario, se registró una elevada mortalidad cuando se expusieron por contacto a la malla tratada durante 72 horas en cajas de dimensiones reducidas (10 cm de diámetro X 3 cm de altura). En ensayos llevados a cabo bajo condiciones más reales de exposición, en un invernadero experimental con jaulas de 25 X 25 X 60 cm de altura, no se produjo ningún efecto en la mortalidad a corto plazo (72 horas) o en los parámetros reproductivos de O. laevigatus y N. tenuis. Finalmente, en ensayos de campo realizados en túneles semi-comerciales (8 m de largo X 6,5 m de ancho X 2,6 m de altura), ni las condiciones ambientales [temperatura, humedad relativa, radiación ultravioleta (UV) y fotosintéticamente activa (PAR)], ni los enemigos naturales, se vieron afectados por la presencia de la malla tratada con bifentrin en el cultivo. Sin embargo, los resultados no fueron concluyentes, debido al bajo establecimiento de los agentes de biocontrol liberados. Por lo tanto, más estudios son necesarios en invernaderos comerciales para confirmar los resultados preliminares de compatibilidad. Además, en este trabajo se han evaluado los efectos letales (mortalidad) y subletales (parámetros reproductivos) de seis modernos insecticidas sobre los chinches depredadores O. laevigatus y N. tenuis, mediante ensayos de laboratorio y persistencia. Los ensayos se realizaron por contacto residual, aplicando los insecticidas a la dosis máxima de campo sobre placas de cristal (laboratorio) o plantas (persistencia). Los productos fitosanitarios se seleccionaron por representar a un grupo de modernos plaguicidas con modos de acción en principio más selectivos para los enemigos naturales que antiguos plaguicidas como organoclorados, oroganofosforados o carbamatos, y por su uso frecuente en cultivos hortícolas donde O. laevigatus y N. tenuis están presentes. Todos ellos están incluidos o en proceso de inclusión en la lista comunitaria de sustancias activas para uso agrícola, Anexo I de la Directiva 91/414/CEE: abamectina y emamectina (avermectinas neurotóxicas, activadoras del canal del cloro), deltametrina (piretroide neurotóxico, modulador del canal del sodio, control positivo), flubendiamida (neurotóxico, modulador del receptor de rianodina), spinosad (naturalito neurotóxico, agonistas/antagonistas del receptor de nicotínico acetilcolina) y spiromesifen (inhibidor de la acetil CoA carboxilasa). El estudio mostró que O. laevigatus fue más susceptible a los insecticidas que N. tenuis. Además, los resultados revelaron que flubendiamida y spiromesifen fueron compatibles con los dos enemigos naturales estudiados, y por tanto se podrían usar en programas de GIP. Por el contrario, los insecticidas abamectina, deltametrina, emamectina y spinosad no fueron selectivos para ninguno de los chinches depredadores. Sin embargo, los estudios de persistencia demostraron que a pesar de que estos insecticidas no proporcionaron selectividad fisiológica, pueden proporcionar selectividad ecológica en algunos casos. Abamectina, deltametrina, emamectina y spinosad podrían ser compatibles con N. tenuis si el enemigo natural es introducido en el cultivo 4 días después de su aplicación. En el caso de O. laevigatus, abamectina, deltametrina y spinosad se clasificaron como persistentes, por lo tanto es necesario completar el estudio con experimentos de semi-campo y campo que determinen si es posible su uso conjunto en programas de GIP. Por otro lado, emamectina podría ser compatible con O. laevigatus si el enemigo natural es introducido en el cultivo 7 días después de su aplicación. Por último, se ha comprobado la selectividad de tres insecticidas aceleradores de la muda (MACs) (metoxifenocida, tebufenocida y RH-5849) sobre O. laevigatus y N. tenuis. Además de realizar estudios para evaluar la toxicidad en laboratorio de los insecticidas por contacto residual e ingestión (principal modo de acción de los MAC´s), se extrajo RNA de los insectos y con el cDNA obtenido se secuenció y clonó el dominio de unión al ligando (LBD) del receptor de ecdisona correspondiente a O. laevigatus (OlEcR-LBD) y N. tenuis (NtEcR-LBD). Posteriormente, se obtuvo la configuración en tres dimensiones del LBD y se estudió el acoplamiento de las moléculas de los tres insecticidas en la cavidad que forman las 12 α-hélices que constituyen el EcR-LBD. En el caso de N. tenuis se debe mencionar que no fue posible la obtención de la secuencia completa del LBD. Sin embargo, se obtuvo una secuencia parcial (hélice 6-hélice 11), que mostró una alta conservación de aminoácidos con respecto a la obtenida en O. laevigatus. Los ensayos de toxicidad mostraron que metoxifenocida, tebufenocida y RH-5849 no produjeron ningún efecto nocivo en ambos depredadores. Además, los estudios de modelado por homología y acoplamiento molecular llevados a cabo con O. laevigatus, también indicaron que los MACs no produjeron ningún efecto deletéreo en este enemigo natural. Por lo tanto, estos compuestos pueden ser aplicados de manera segura en programas de GIP en los cuales O. laevigatus y N. tenuis estén presentes. ABSTRACT The new pesticide legislation on pest control is aimed at integrated pest management (IPM). These programs are based on the most environmentally sustainable approaches, where biological, physical control and other non-chemical methods are the cornerstone. However, selective pesticides are often required for pest management on horticultural crops. Therefore, the main goal of this study is to provide knowledge to improve pest control on horticultural crops through the integration of three strategies: biological, physical and chemical. Firstly, the effects of insecticide treated nets (bifenthrin) were evaluated in different laboratory, greenhouse and field experiments on the natural enemies Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) (predator of thrips), Nesidiocoris tenuis (Reuter) (Hemiptera: Miridae) (predator of whiteflies and Tuta absoluta (Meirick) (Lepidoptera: Gelechiidae)), and other biocontrol agents commonly used on protected horticultural crops. These types of nets have been successfully used in medical entomology to control mosquito malaria vectors, and work is currently being done on their use as exclusion barriers and as a direct method of pest control in agriculture. In experiments made under laboratory conditions, O. laevigatus and N. tenuis were not able to detect the presence of bifenthrin in a dual-choice test. Furthermore, no shortterm mortality (72 hours) was recorded on both predatory bugs. In contrast, a high mortality rate was found when they were exposed by contact to the bifenthrin-treated net for 72 hours in small cages (10 cm diameter X 3 cm high). In assays carried out under more realistic conditions of exposure, in an experimental greenhouse with cages of 25 X 25 X 60 cm high, short-term mortality (72 hours) and reproductive parameters were not affected. Lastly, in field experiments carried out in semi-commercial tunnels (8 m long X 6.5 m width X 2.6 m high), neither environmental conditions [temperature, relative humidity, ultraviolet (UV) and photosynthetically active radiation (PAR)] nor natural enemies were affected by the presence of the bifenthrin-treated net on the crop. However, results were not conclusive, mainly due to a low settlement of the released biocontrol agents, and further studies are needed in commercial greenhouses to confirm our preliminary results of compatibility. Secondly, the lethal (mortality) and sublethal effects (reproductive parameters) of six modern pesticides on the predatory bugs O. laevigatus and N. tenuis has been evaluated through laboratory and persistence experiments. Trials were carried out by residual contact, applying the insecticides to the maximum field recommended concentration on glass plates (laboratory) or plants (persistence). Insecticides were chosen as representatives of modern pesticides with a more selective mode of action on natural enemies than organochlorine, organophosphorus and carbamate insecticides. Moreover, they were also chosen because of their frequent use on horticultural crops where O. laevigatus and N. tenuis are present. All of them have been included or have been requested for inclusion in the community list of active substances on the agricultural market, Annex I of the European Directive 91/414/EEC: abamectin and emamectin (neurotoxic avermectins, chloride channel activators), deltamethrin (neutotoxic pyrethroid, sodium channel modulator, positive commercial standard), flubendiamide (neurotoxic, rianodine receptor modulator), spinosad (neurotoxic naturalyte, nicotinic acetylcholine receptor allosteric activator) and spiromesifen (inhibitors of acetyl CoA carboxylase). The study showed that O. laevigatus was more susceptible to all the studied pesticides than N. tenuis. In addition, the research results indicated no impact of flubendiamide and spiromesifen on the two natural enemies studied under laboratory conditions. Consequently, both pesticides are candidates to be included in IPM programmes where these biocontrol agents are present. On the other hand, abamectin, deltamethrin, emamectin and spinosad were not selective for both predatory bugs in laboratory experiments. However, persistence test demonstrated that in spite of the lack of physiological selectivity, these pesticides can provide ecological selectivity in some cases. Abamectin, deltamethrin, emamectin and spinosad could be compatible with N. tenuis if the mirid bug is released 4 days after the insecticide treatment on the crop. With regard to O. laevigatus, abamectin, deltamethrin and spinosad were classified as persistent in our assays, thus the study should be completed with semi-field and field experiments in order to ascertain their possible joint use in IPM programs. In contrast, emamectin could be compatible with O. laevigatus if the pirate bug is released 7 days after the insecticide treatment on the crop. Finally, the selectivity of three moulting accelerating compounds (MACs) (methoxyfenozide, tebufenozide and RH-5849) has also been evaluated on O. laevigatus and N. tenuis. In addition to laboratory experiments to evaluate the toxicity of the insecticides by residual contact and ingestion, molecular approaches were used as well. RNA of both insects was isolated, cDNA was subsequently synthesized and the complete sequence of the ligand binding domain (LBD) of the ecdysone receptor of O. laevigatus (OlEcR-LBD) and N. tenuis (NtEcR-LBD) were determined. Afterwards, the three dimensional structure of LBD was constructed. Finally, the docking of the insecticide molecules in the cavity delineated by the 12 α-helix that composed the EcRLBD was performed. In the case of N. tenuis, it should be noted that in spite of intensive efforts, we did not manage to complete the sequence for the LBD.However, a partial sequence of the LBD was obtained (helix 6-helix 11), and a strong conservation between the amino acids of N. tenuis and O. laevigatus was observed. Results showed no biological activity of methoxyfenozide, tebufenozide and RH-5849, on both predatory bugs. Moreover, modeling of the OlEcR-LBD and docking experiments also suggested that MACs were devoid of any deleterious effect on O. laevigatus. Therefore, our results indicate that these compounds could be safely applied in IPM programs in which O. laevigatus and N. tenuis are present.

Artificial diets for the production of natural enemies (predators and parasitoids) of greenhouse pest insects

The aim of this contract was to finalise in vitro rearing on artificial diets of entomophagous insects useful to control insect pests mainly in greenhouses through an analytical and rational approach. The work focuses on the development and optimisation of artificial diets for one coccinellid (Harmonia axyridis), two heteropteran predators (Orius laevigatus, Dicyphus tamaninii), and a braconid parasitoid of aphids (Aphidius ervi). The definition of the artificial diets was based on biochemical analyses of their natural food (aphids) or substitution food for the predators (Ephestia kuehniella eggs). Reliable quality control parameters of the entomophages produced were used in order to adjust dietary composition and formulation of the different diets tested.

Efeitos do plantio direto e da consorciação soja-milho sobre inimigos naturais e pragas

Procurou-se avaliar os efeitos do plantio direto e da consorciação soja (Glycine max (L.) Merrill) e milho (Zea mays L.) sobre pragas e inimigos naturais. Os tratamentos constituíram um fatorial 3 x 2 (monocultura de soja, monocultura de milho, consorciação soja-milho x plantio direto, plantio convencional), em blocos casualizados. Os insetos foram amostrados pelo método do pano, rede entomológica, procura visual e armadilha de sucção. Entre os insetos-pragas do milho, Maecolaspis assimilis ocorreu em maior número no sistema de plantio convencional; o mesmo ocorreu com os predadores Cycloneda sanguinea e Doru sp. Por outro lado, M. assimilis e o predador Toxomerus sp. foram mais numerosos na monocultura de milho em relação à cultura do milho consorciado com soja. Dos insetos-pragas da soja, destacaram-se pelo maior número Anticarsia gemmatalis e Diabrotica gracilenta, no sistema de plantio convencional, e o mesmo aconteceu com a espécie da família Trichogrammatidae, enquanto as espécies da família Eulophidae foram mais numerosas na soja sob sistema de plantio direto. Na soja consorciada com milho foi maior o número de insetos-pragas Megalotomus sp. e Maecolaspis sp. e dos inimigos naturais Geocoris sp., Lebia concina, Orius sp., Braconidae e Scelionidae.