Dispersal of Cotesia flavipes in sugarcane field and implications for parasitoid releases

Diatraea saccharalis Fabr. (Lepidoptera: Crambidae) is a major sugarcane pest in Brazil. The management of infested areas is based on the release of Cotesia flavipes (Cameron) (Hymenoptera: Braconidae), a parasitoid of D. saccharalis larvae, but there are doubts about the effectiveness of C. flavipes, primarily regarding its rate of dispersal in sugarcane fields. Thus, the objective of this study was to evaluate the dispersal of C. flavipes in a sugarcane field and suggest a release method that provides higher parasitoid efficiency. The study was carried out in four areas of approximately 1 ha, in which stalk pieces containing 20 D. saccharalis larvae were distributed in a rectangular grid, and 12,000 C. flavipes adults were released at four points, that were 50 m apart and 25 m from the field border. Three days later, the D. saccharalis larvae were recovered and kept in the laboratory until they reached pupal stage or C. flavipes emergence. Parasitism varied from 13.2% to 42.8%. The random distribution of parasitized larvae was found in one assay. In three areas, the parasitized larvae showed an aggregated distribution, with a range of 15 to 25 m. Since the parasite's success is directly linked to parasitoid dispersion, it would be interesting to move the release points to 30 m from each other because the dispersal may happen in a 15 m radius.

Side Effects of Neem Oil on the Midgut Endocrine Cells of the Green Lacewing Ceraeochrysa claveri (Navas) (Neuroptera: Chrysopidae)

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

Métodos para avaliar a liberação e o parasitismo de Cotesia flavipes (Cameron, 1891) (Hymenoptera: Braconidae) em cana-de-açúcar

Pós-graduação em Agronomia (Entomologia Agrícola) - FCAV

Resposta de Cotesia kariyai (Watanabe) e Cotesia flavipes (Cameron) (Hymenoptera: Braconidae) a voláteis induzidos por herbivoria

Pós-graduação em Agronomia (Entomologia Agrícola) - FCAV

Desenvolvimento de nova embalagem para comercialização, transporte e liberação do parasitoide Cotesia flavipes (Cameron, 1891) (Hymenoptera: Braconidae)

Pós-graduação em Agronomia (Entomologia Agrícola) - FCAV

Interaction between N-fertilizer and water availability on borer-rot complex in sugarcane

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeito da alimentação do adulto na qualidade do endoparasitoide larval da broca comum da cana-de-açúcar

Aiming qualitative improvements in the mass rearing of Cotesia flavipes (Cameron, 1891) (Hymenoptera: Braconidae) in laboratory, tests were conducted with adults feed before parasitism on Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) larvae, using 14 artificial diets based on sucrose, brewer yeast and honey. The design was completely randomized with 15 treatments and five replications. The biological parameters evaluated in F1 and F2 generations were: number of adults emerged; sex ratio; adult emergence percentage; weight of pupae mass; and male and female longevity. The highest average of emerged adults were 85 in diet with sucrose 5% for F1 generation, and 91 in diet with sucrose 5% + yeast brewer 2,5% for F2; the smaller average of adult emergency were 63.60 in diet with honey 5% + yeast brewer 2.5 % for F1 generation and 66.80 in diet with honey 5% for F2. Diets that produced more females were honey 5% + yeast brewer 2.5%, showing the sex ratio of 0.82 for F1 generation, and sucrose 10% + 2.5% yeast brewer 2.5% (0.75) for F2, while sucrose 10% + yeast brewer 2.5% (0.60 for F1) and honey 2.5% (0.59) provided fewer females for F2 generation. The highest percentage of adult emergence were 93.90% with the diet containing sucrose 2.5% + yeast brewer 2.5%) and 93.89 % with honey 5% + yeast brewer 2.5%, and the lowest 81.71% (honey 5% + yeast brewer 2.5%) for F1 and 78.96 % (honey 5%) to F1 and F2, respectively. The diet with Mel 2.5% produced greater weight of pupae mass. The adult longevity did not differ significantly for all diets in F1 and F2 generations. Diets containing 5% or 10% of sucrose or honey provide, in general, improve the qualitative development of C. flavipes in mass rearing.

Development of a New Container for Storage and Release of the Parasitoid Cotesia flavipes (Hymenoptera: Braconidae)

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

Cytotoxic effects of neem oil in the midgut of the predator Ceraeochrysa claveri

Studies of morphological and ultrastructural alterations in target organs have been useful for evaluating the sublethal effects of biopesticides regarded as safe for non-target organisms in ecotoxicological analyses. One of the most widely used biopesticides is neem oil, and its safety and compatibility with natural enemies have been further clarified through bioassays performed to analyze the effects of indirect exposure by the intake of poisoned prey. Thus, this study examined the cellular response of midgut epithelial cells of the adult lacewing, Ceraeochrysa claveri, to neem oil exposure via intake of neem oil-contaminated prey during the larval stage. C. claveri larvae were fed Diatraea saccharalis eggs treated with neem oil at concentrations of 0.5%, 1% and 2% throughout the larval stage. The adult females obtained from these treatments were used at two ages (newly emerged and at the start of oviposition) in morphological and ultrastructural analyses. Neem oil was found to cause pronounced cytotoxic effects in the adult midgut, such as cell dilation, emission of cytoplasmic protrusions, cell lysis, loss of integrity of the cell cortex, dilation of cisternae of the rough endoplasmic reticulum, swollen mitochondria, vesiculated appearance of the Golgi complex and dilated invaginations of the basal labyrinth. Epithelial cells responded to those injuries with various cytoprotective and detoxification mechanisms, including increases in cell proliferation, the number of calcium-containing cytoplasmic granules, and HSP 70 expression, autophagic processes and the development of smooth endoplasmic reticulum, but these mechanisms were insufficient for recovery from all of the cellular damage to the midgut. This study demonstrates that neem oil exposure impairs the midgut by causing sublethal effects that may affect the physiological functions of this organ, indicating the importance of studies of different life stages of this species and similar species to evaluate the safe and compatible integrated use of biopesticides.

Produção semissólida de metarhizium anisopliae e beauveria bassiana em diferentes substratos e efeito da radiação ultravioleta e da temperatura sobre propágulos desses entomopatógenos

The present study was aimed to evaluate different (semi-solid) media for the production of Metarhizium anisopliae and Beauveria bassiana propagules, and to evaluate the tolerance of these propagules to ultraviolet radiation and temperature. The experiments were performed at the Biological Control Laboratory of the Instituto Biológico at Campinas, São Paulo, Brazil. For both fungi, 6 repetitions were performed for each of the 17 treatments: corn starch, full rice, parboiled rice, type-1 rice, type-2 rice, oat flakes, canjiquinha [grits], wheat flour, raw cassava flour, yellow corn flour, special wheat flour, corn flour, corn in grains, cassava starch, soy in grains, crushed wheat, and turf. The viability analysis was done in plastic plates containing BDA. For the bioassays involving exposure to ultraviolet light and temperature, BDA was also used for viability analysis, and each treatment was exposed to the UV radiation for 0, 25 and 50 seconds, the temperature exposure being at 20, 25, 30 and 35º C. Using a Potter tower, 2 mL of fungus suspension from each treatment was inoculated to the Diatraea saccharalis caterpillars. Regarding the sporulation, the largest concentrations of M. anisopliae and B. bassiana were found for the treatments with parboiled rice, type-1 rice, type-2 rice, yellow corn flour, corn flour and crushed wheat. The viability of all treatments was superior to 94.00%. Also, the longer the duration of the exposition to the UV, the smaller the number of fertile conidia. At 35o C, a significant loss of conidia viability was observed, and all the treatments presented some level of virulence.

Avaliação da capacidade de renovação do epitélio do mesêntero e seus efeitos no potencial reprodutivo de fêmeas adultas de Ceraeochrysa claveri (Neuroptera: Chrysopidae) alimentadas durante a fase larval com óleo de nim (azadirachta indica A. JUSS)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização e comparação de Cotesia flavipes (Hymenoptera: braconidae) em situação de criação massal utilizando microssatélites e morfologia

Pós-graduação em Agronomia (Entomologia Agrícola) - FCAV

Biological and behavioral parameters of the parasitoid Cotesia flavipes (Hymenoptera: Braconidae) are altered by the pathogen Nosema sp (Microsporidia: Nosematidae)

A major issue for mass rearing of insects concerns sanitary conditions and disease. Microsporidian infection (Nosema sp.) in laboratory colonies of Diatraea saccharalis (Fabr.) (Lepidoptera: Crambidae), used in producing the parasitoid. Cotesia flavipes Cameron (Hymenoptera: Braconidae), is representative of the problems faced by growers and industry. Although C. flavipes has been produced for several years in Brazil for biological control of D. saccharalis, we have only recently observed that the parasitoid becomes infected when developing inside hosts infected with Nosema sp. We assessed the effects of Nosema sp. on C. flavipes, including the ability to locate and select hosts, and evaluated pathogen transmission. Third instar larvae of D. saccharalis were inoculated with Nosema sp. spores at different concentrations and were parasitized when larvae reached fifth instar. Heavily infected D. saccharalis larvae did not support parasitism. Parasitoids that developed in infected D. saccharalis larvae exhibited increased duration of larval and pupal stages, decreased adult longevity and number of offspring, and reduced tibia size compared to parasitoids developing in uninfected D. saccharalis larvae. Infection by Nosema sp. reduced the ability of the C. flavipes parasitoid to distinguish between volatiles released by the sugarcane infested by healthy larvae and pure air. Uninfected parasitoids preferred plants infested with uninfected hosts. But infected C. flavipes did not differentiate between uninfected hosts and those infected with Nosema sp. The pathogen is transmitted from host to parasitoids and parasitoids to hosts. Pathogenic effects of the microsporidium in C. flavipes are sufficiently severe to justify disease management efforts, particularly considering the importance of C. flavipes as a biological control agent in sugarcane. (C) 2012 Elsevier Inc. All rights reserved.

EVALUATION OF IN VITRO AND IN VIVO EFFECTS OF SEMIPURIFIED PROTEINASE INHIBITORS FROM THEOBROMA SEEDS ON MIDGUT PROTEASE ACTIVITY OF LEPIDOPTERAN PEST INSECTS

We have characterized in vitro and in vivo effects of trypsin inhibitors from Theobroma seeds on the activity of trypsin- and chymotrypsin-like proteins from Lepidopteran pest insects. The action of semipurified trypsin inhibitors from Theobroma was evaluated by the inhibition of bovine trypsin and chymotrypsin activities determined by the hydrolysis of N-Benzoyl-DL-Arginine-p-Nitroanilide (BAPA) and N-Succinyl-Ala-Ala-Pho-Phe p-Nitroanilide (S-(Ala)2ProPhe-pNA). Proteinase inhibitor activities from Theobroma cacao and T. obovatum seeds were the most effective in inhibiting trypsin-like proteins, whereas those from T. obovatum and T. sylvestre were the most efficient against chymotrypsin-like proteins. All larvae midgut extracts showed trypsin-like proteolytic activities, and the putative trypsin inhibitors from Theobroma seeds significantly inhibited purified bovine trypsin. With respect to the influence of Theobroma trypsin inhibitors on intact insects, the inclusion of T. cacao extracts in artificial diets of velvet bean caterpillars (Anticarsia gemmatalis) and sugarcane borer (Diatraea saccharalis) produced a significant increase in the percentage of adult deformation, which is directly related to both the survival rate of the insects and oviposition.

Sugarwin: A Sugarcane Insect-Induced Gene with Antipathogenic Activity

In sugarcane fields, colonization of the stalk by opportunistic fungi usually occurs after the caterpillar Diatraea saccharalis attacks the sugarcane plant. Plants respond to insect attack by inducing and accumulating a large set of defense proteins. Two homologues of a barley wound-inducible protein (BARWIN), sugarcane wound-inducible proteins SUGARWIN1 and SUGARWIN2, have been identified in sugarcane by an in silico analysis. Antifungal properties have been described for a number of BARWIN homologues. We report that a SUGARWIN:green fluorescent protein fusion protein is located in the endoplasmic reticulum and in the extracellular space of sugarcane plants. The induction of sugarwin transcripts occurs in response to mechanical wounding, D. saccharalis damage, and methyl jasmonate treatment. The accumulation of transcripts is late induced and is restricted to the site of the wound. Although the transcripts of sugarwin genes were strongly increased following insect attack, the protein itself did not show any effect on insect development; rather, it altered fungal morphology, leading to the apoptosis of the germlings. These results suggest that, in the course of evolution, sugarwin-encoding genes were recruited by sugarcane due to their antipathogenic activity. We rationalize that sugarcane is able to induce sugarwin gene expression in response to D. saccharalis feeding as a concerted plant response to the anticipated invasion by the fungi that typically penetrate the plant stalk after insect damage.