Avaliação da ação bioinseticida de SBTI e vicilina de Erythrina velutina em enzimas digestivas e membrana peritrófica de larvas de Plodia interpunctella (Lepidoptera: Pyralidae)

Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests

Combinación de diferentes estrategias de aplicación de hongos entomopatógenos para el control de Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) y Bemisia tabaci (Gennadius) (Hemiptera:Aleyrodidae)

Los ascomicetos mitospóricos entomopatógenos, que actúan por vía tegumentaria, han sido utilizados para el control microbiano de plagas mediante distintas estrategias que permiten el contacto de sus conidios con los insectos diana. Sin embargo, se conocen nuevos aspectos sobre su función ecológica tales como su capacidad secretora de compuestos con actividad insecticida, así como su sorprendente comportamiento como endófitos, que complementan su empleo clásico y que pueden pantag ermitir el desarrollo de nuevas estrategias de control de plagas. El empleo conjunto de estos hongos y sus extractos para el control de un insecto polífago de importancia mundial Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) es abordado en el capítulo II, donde se pone de manifiesto la existencia de variación intra- e interespecífica en la virulencia y estrategias patogénicas de las cepas de Beauveria bassiana y Metarhizium brunneum evaluadas frente a larvas del noctuido. La aplicación conjunta de cepas con distintas estrategias patogénicas junto con sus extractos a larvas del lepidóptero tuvo efecto aditivo, y alguno antagónico, de lo que depende su empleo conjunto. Los capítulos III y IV revelan la existencia de colonización endofítica transitoria tras la aplicación foliar de suspensiones de conidios de cepas de las especies mencionadas en alfalfa, tomate y melón, así como el impacto de esta colonización sobre dos fitófagos con diferentes hábitos de alimentación, uno masticador S. littoralis y otro picador-suctor Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). El capítulo III muestra la contribución aditiva de la mortalidad causada por la alimentación de larvas de S. littoralis a expensas de material vegetal colonizado endofíticamente por cepas de B. bassiana y M. anisopliae con la debida al tratamiento tópico de las mismas con las suspensiones fúngicas. El origen de la mortalidad iniciada a través de la vía digestiva en larvas de este lepidóptero permanece incierto para las cepas de B. bassiana, pero podría estar asociado a la presencia en la planta de metabolitos fúngicos en las cepas de M. brunneum como denota la presencia de destruxina (dtx) A en el 11,0% de los cadáveres. El capítulo IV revela que cepas de ambas especies fúngicas pueden iniciar ciclos de infección en insectos picadores-suctores cuando se alimentan a expensas de sustrato vegetal colonizado endofíticamente, si bien, ambas especies presentan estrategias diferentes. Así, B. bassiana muestra una gran capacidad para colonizar el melón, incluso con efecto traslaminar, que causa la infección de ninfas de B. tabaci por contacto con el tegumento. Sin embargo, la mortalidad de las mismas causada por M. brunneum, de crecimiento mucho más localizado en la hoja, está relacionada una vez mas con la presencia de dtx A en el 43,0% de los cadáveres. Estos resultados deben ser considerados para la evaluación del impacto real de los tratamientos con hongos entomopatógenos, y abren nuevas vías en el control de plagas.

Resistance of Bt transgenic maize to lesser cornstalk borer (Lepidoptera: Pyralidae).

To determine if Bt maize seedlings are protected against lesser cornstalk borer damage, Bt hybrids at the 3 and 4 leaf stages were tested under temperatures between 20-32oC and artificial infestation. A high level of resistance was reported in all Bt maize as expressed by larval survival, larval weight, damage score, and number of surviving plants. All Bt maize protected plants against lesser cornstalk borer damage. Also, the resistance present in the Bt maize was not affected by daytime temperature.

Development and reproduction of Olla v-nigrum (Coleoptera: Coccinellidae) fed Anagasta kuehniella (Lepidoptera: Pyralidae) eggs supplemented with an artificial diet.

2013

Nomuraea rileyi (Hypocreales: Clavicipitaceae) in Helicoverpa armigera (Lepidoptera: Noctuidae) Larvae in Brazil.

2015

Comparison of biology between Helicoverpa zea and Helicoverpa armigera (Lepidoptera: Noctuidae) reared on artificial diets.

The objective of this study was to compare growth and development of Helicoverpa zea Boddie and Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) larvae that were feeding on an artificial diet. Neonate larvae of H. zea and H. armigera were collected in maize fields cultivated at the Brazilian Agricultural Research Corporation, Embrapa Maize & Sorghum, in Sete Lagoas (Minas Gerais) and in farmers? cotton fields in Luís Eduardo Magalhães (Bahia), respectively. Bioassays were conducted in the laboratory using individual larvae in 50 mL plastic cups fed a white bean?based artificial diet and maintained at a temperature of 26 ± 2 °C and a relative humidity of 47 ± 10%. The following larval biological parameters were evaluated: number and duration of instars, survival of larval instars and pupae, larval biomass, larval head capsule size, and larval length. An adaptation index was computed for comparison of development of the 2 species. Significant differences were observed between the species for all variables except for the number of instars and pupal survival. Although H. armigera larvae developed faster and were smaller than H. zea larvae, the diet tested can be considered adequate for rearing both species in the laboratory.

Avaliação da potencial migração de Helicoverpa armigera (Lepidoptera: Noctuidae) por massas de ar para áreas produtoras de cultivos hospedeiros do Estado de São Paulo.

Resumo Helicoverpa armigera (Hübner, 1808) (Lepidoptera: Noctuidae) é um inseto exótico, altamente polífago, que foi detectado no Brasil em 2013 causando sérios danos em cultivos de soja, milho e algodão, localizados em áreas estaduais específicas das regiões Nordeste e Centro-Oeste com a presença do bioma Cerrado. Após o período de um ano o inseto foi identificado em várias regiões do País. A literatura indica que o inseto adulto apresenta características físicas que favorecem sua grande capacidade de voo, tanto em distância (100-1000 km) quanto a elevadas altitudes (maiores que 1,5 km acima do nível do solo), aumentando o seu potencial para dispersão de longo alcance, principalmente por voos migratórios das áreas já atacadas. Por essa razão, as massas de ar brasileiras podem atuar como potenciais facilitadoras à atividade migratória do inseto a longas distâncias para as áreas de maior concentração de seus cultivos hospedeiros preferenciais, resultante da atração natural do inseto aos fatores químicos delas decorrentes. Desse modo, devem ser consideradas em avaliações de áreas mais propícias à presença de H. armigera para orientar monitoramentos e estratégias de manejo locais. Este trabalho avaliou a potencial migração de H. armigera por massas de ar para áreas produtoras de cultivos hospedeiros do estado de São Paulo, considerando seus municípios produtores de algodão, soja, milho, tomate, feijão, café e laranja; as principais massas de ar brasileiras; a presença de grandes áreas com cultivos de eucalipto e pinus (outras potenciais barreiras físicas ao inseto); e a localização do bioma Cerrado paulista. Informações das áreas produtoras e das áreas plantadas por município de 2008, 2012 e 2014, disponibilizadas pelo IBGE, foram utilizadas em planos de informação georreferenciados, elaborados em ArcGis 10.3, para identificar a localização de suas respectivas grandes áreas em cada ano. O cruzamento georreferenciado também resultou mapas das áreas plantadas em 2014 com todos os cultivos citados, indicando as de maior potencial de infestação/reinfestações de H. armigera no estado de São Paulo, para orientar monitoramentos e estratégias de Manejo Integrado de Pragas - MIP locais. Abstract Helicoverpa armigera (Hübner, 1808) (Lepidoptera: Noctuidae) is an exotic insect highly polyphagous, which was detected in Brazil in 2013 causing serious damage to soybean, corn and cotton crops located in specific state areas in the northeast and center-west regions with the presence of Cerrado Biome (similar to the Savannah). After a oneyear period, the insect was detected in several regions of the country. Foreign literature indicates that the adult phase of the insect shows certain physical characteristics which favor its great flight capacity, even related to reach long distances (100-1000 Km) as many as to reach high altitudes (overtaken 1.5 Km above the ground level), increasing its potential of farthest dispersion, mainly by migratory flights from the areas already attacked. Therefore, the Brazilian air masses may act as potentially enhance the insect's farthest migratory activity to areas of greater concentration of its preferential host crops due to the insect's natural attraction to their chemical factors. Thus, migration potential of H. armigera by air masses toward preferential host crop areas must be considered in evaluations of most favorable areas to the presence of the insect to guide monitoring and local management strategies. The present work evaluated the H. armigera migration potential by air masses toward producing areas of host crops in São Paulo state, Brazil, considering its municipalities producing cotton, soybean, corn, tomato, bean, coffee, orange, eucalyptus and pine trees, as well as the main Brazilian air masses and the presence of Cerrado Biome in the state. Information of producing and of planting areas with the annual crops by municipalities in 2008, 2012 and 2014, provided by IBGE, were used in cross-maps made using ArcGis 10.3, in order to identify the location of their respective greater areas in each year. Cross-maps also provided results indicating the planting areas in 2014 with all crops already mentioned, as well as the potential areas to infestation/reinfestations of H. armigera in São Paulo State, in order to guide monitoring and local strategies of IPM.

Natural parasitism of lepidopteran eggs by Trichogramma species (Hymenoptera: Trichogrammatidae) in agricultural crops in Minas Gerais, Brazil.

The genus Trichogramma Westwood (Hymenoptera: Trichogrammatidae) includes insect egg parasitoids that are widely used throughout the world as control agents of pest insects. The aim of this study was to identify the species of Trichogramma naturally associated with the eggs of lepidopteran pests of the following agricultural and horticultural crops: collards, Brassica oleracea L. (Brassicales: Brassicaceae); papaya, Carica papaya L. (Capparales: Caricaceae); tomato, Lycopersicon esculentum Mill. (Solanales: Solanaceae); cassava, Manihot esculenta Crantz (Malpighiales: Euphorbiaceae); banana, Musa sp. L. (Zingiberales: Musaceae); passion fruit, Passiflora sp. Degener (Malpighiales: Passifloraceae); sugarcane, Saccharum sp. L. (Poales: Poaceae); and corn (maize), Zea mays L. (Poales: Poaceae); and an invasive species (Sodom?s apple milkweed, Calotropis procera Aiton; Gentianales: Apocynaceae) in the semiarid region of Minas Gerais, Brazil. We report natural parasitism by Trichogramma in eggs of Agraulis vanillae vanillae (L.) (Lepidoptera: Nymphalidae), Antichloris eriphia F. (Lepidoptera: Arctiidae), Danaus sp. (L.) (Lepidoptera: Nymphalidae), Diatraea saccharalis F. (Lepidoptera: Crambidae), Erinnyis ello L. (Lepidopera: Sphingidae), and Protambulyx strigilis L. (Lepidopera: Sphingidae). In total, 2,242 specimens of Trichogramma were obtained, belonging to the species T. pretiosum Riley, T. manicobai Brun, Moraes & Soares, T. marandobai Brun, Moraes & Soares, and T. galloi Zucchi. These species of Trichogramma may be candidates for biological control programs of lepidopteran pests in the semiarid region of Minas Gerais and in other semiarid regions.

Avaliação de inseticidas biológicos e fisiológicos para controle da 'grande lagarta' Physocleora dimidiaria (Lepidoptera: geometridade).

2009

Species from the Heliothinae complex (Lepidoptera: noctuidae) in Tucuman, Argentina, an update of geographical distribution of Helicoverpa armigera.

The Heliothinae complex in Argentina encompasses Helicoverpa gelotopoeon (Dyar), Helicoverpa zea (Boddie), Helicoverpa armigera (Hu ̈ bner), and Chloridea virescens (Fabricius). In Tucum an, the native species H. gelotopoeon is one of the most voracious soybean pests and also affects cotton and chickpea, even more in soybean-chickpea succession cropping systems. Differentiation of the Heliothinae complex in the egg, larva, and pupa stages is difficult. Therefore, the observation of the adult wing pattern design and male genitalia is useful to differentiate species. The objective of this study was to identify the species of the Heliothinae complex, determine population fluctuations of the Heliothinae complex in soybean and chickpea crops using male moths collected in pheromone traps in Tucuman province, and update the geographical distribution of H. armigera in Argentina. The species found were H. gelotopoeon, H. armigera, H. zea , and C. virescens. Regardless of province, county, crop, and year, the predominant species was H. gelotopoeon . Considering the population dynamics of H. gelotopoeon and H. armigera in chickpea and soybean crops, H. gelotopoeon was the most abundant species in both crops, in all years sampled, and the differences registered were significant. On the other hand, according to the Sistema Nacional Argentino de Vigilancia y Monitoreo de Plagas (SINAVIMO) database and our collections, H. armigera was recorded in eight provinces and 20 counties of Argentina, and its larvae were found on soybean, chickpea, sunflower crops and spiny plumeless thistle (Carduus acanthoides). This is the first report of H. armigera in sunflower and spiny plumeless thistle in Argentina.

Mortalidade natural de lagartas de Chrysodeixis includens (Walker, 1857) (Lepidoptera: Noctuidae) em lavouras de soja na região de Dourados - MS.

2015

Flutuação populacional de machos adultos de Helicoverpa spp. (Lepidoptera; noctuidae) em Mato Grosso do Sul.

2015

Estratégias de acasalamento e qualidade do espermatóforo em heliconius (Lepidoptera, Nymphalidae)

Butterflies are insects known, in a variety of environments and for easy visual identification. The adult form may be frequently found in flowers looking for nectar. However, for many species of Heliconius (Lepidoptera, Nymphalidae) to visit the flower also represents the collecting of pollen, an important source of protein for adults. The protein obtained from the pollen allows the maintenance of physiological processes that increase the performance of the individual, promoting greater longevity and egg production. For males, proteins can also be part in your investment in reproductive success and fitness of offspring through a nutritional contribution that is transferred to the female in the act of mating as a nuptial present. It is known that this protein contains essential to the performance of the female oviposition, however the proportion of content and specific importance to the monogamous and polygamous species is not known yet. Whereas the species studied in this work have different patterns of mating in the strategy was to verify a significant difference in the quality of the spermatophore, and H. erato and H. melpomene, on the amount of protein present in this structure, indicating a difference in investment between the male reproductive strategies

The influence of nitrogen fertilization on growth, yield, nitrate and oxalic acid concentration in purslane ( Portulaca oleracea )

Purslane (Portulaca oleracea) is widely used for culinary purposes throughout Mediterranean region, and the interest in this plant increased due to it being a source of bio-protective compounds, such as fatty acids and antioxidants. However, the use of purslane could be limited by accumulation of high levels of compounds harmful to human health, such as nitrate and oxalic acid. The main objective of present study was to evaluate the influence of nitrogen fertilization on growth and yield parameters and on nitrate and oxalic acid concentrations in leaves and stems. Plants of golden-leafed purslane of sativa subspecies were grown in styro-foam boxes with substrate and fertilized two times per week during four weeks with ammonium-nitrate solution (16.9% NO3--N and 17.6% NH4+-N), for testing of four nitrogen levels (0, 30, 60 and 90 kg N ha-1). Plant growth, yield, nitrate and oxalic acid concentrations were significantly affected by nitrogen application. The best quantity/quality ratio was achieved at fertilization level of 60 kg N ha-1, which gave a yield of 5.1 kg m-2 FW, while nitrate concentration was 48.98 and 43.90 mg g-1 DW in leaf and stem, respectively, and oxalic acid concentration was 1.27 and 0.55 mg g-1 DW, in leaf and stem, respectively: values which are not harmful for consumer health.

Species richness and composition of butterfly assemblages (Lepidoptera:Rhopalocera) along the altitudinal gradient of Serra da Estrela

Os gradientes altitudinais são importantes modelos em ecologia pois permitem, numa escala reduzida, estudar a influência da variação de diversas variáveis ambientais nos padrões de biodiversidade existentes. Estes padrões são influenciados por diversos factores, desde variáveis climáticas às relacionadas com a perturbação humana. Nesta tese é estudada a variação na composição das comunidades de borboletas ao longo do gradiente altitudinal na Serra da Estrela e são testadas duas hipóteses sobre os padrões de riqueza específica ao longo do gradiente altitudinal: a regra de Rapoport e o “Mid Domain Effect” (MDE). Foram assinaladas 70 espécies de borboletas neste estudo e constatou-se que a variação na riqueza de espécies de borboletas apresenta um padrão “mid peak” apoiando o MDE. Verificou-se também que as comunidades de borboletas do mesmo nível altitudinal são mais semelhantes entre si independentemente da vertente em que se encontram e são também mais semelhantes com as comunidades dos níveis altitudinais adjacentes. Finalmente, a análise da distribuição de algumas espécies ameaçadas de borboletas torna evidente a necessidade de realização de mais estudos sobre a biologia da conservação deste grupo de animais em Portugal; Abstract: Altitudinal gradients are important ecological models because they allow us to study the influence of diverse environmental variables in the existing biodiversity patterns. These patterns are influenced by various factors, from climatic variables to variables related to human disturbance. In this thesis we studied the variations in the composition of the butterfly assemblages along the altitudinal gradient of Serra da Estrela and we tested two hypotheses regarding the species richness patterns along the altitudinal gradient: Rapoport’s rule and the Mid Domain Effect (MDE). In this study 70 butterfly species were marked recorded and it was observed that the butterfly species richness presents a mid-peak pattern supporting the MDE. It was also noted that the butterfly assemblages in the same altitudinal level show more similarity within themselves regardless of the slope they are found in, and are also more similar to the assemblages present in the adjacent altitudinal levels. Finally, the distribution of some threatened butterfly species was analyzed and it became evident the need to do more studies regarding the conservation biology of this animal group in Portugal.