Nilton José Sousa

Conidia Production of Beauveria sp. by Solid-State Fermentation for Biocontrol of Ilex paraguariensis Caterpillars

Conidia production ofBeauveria sp. strain LAG by solid-state fermentation (SSF) using blends of agro-industrial residues (residual potatoes and sugar-cane bagasse) was optimized with respect to cultivation conditions and the composition of substrate mixture in Erlenmeyer flasks and column-type bioreactor. With a blend of 60 % residual potatoes and 40 % sugar-cane bagasse the optimum conditions achieved were: incubation temperature 26 °C, initial substrate pH 6, inoculum concentration 107 conidia per g substrate; optimal initial moisture of the substrate was 70 % for Erlenmeyer flasks, in column-type bioreactor (with forced aeration) the optimal initial moisture of the substrate was 65 % with airflow of 60 mL/min. The highest production (1.07 × 1010 conidia per g dry substrate) was achieved after a 10-d fermentation. The conidia were used in laboratory assays againstThelosia camina andHylesia sp., caterpillars that are serious pests of mate plants. The mortality ofT. camina was >90 % 10 d after spraying caterpillars with 1 mL conidia suspension at a concentration 105–108/mL. ForHylesia sp., the mortality was 70 %, 7 d after immersion in the conidia suspension containing 108 conidia per mL. Therefore, theBeauveria sp. LAG can be considered to be an important biocontrol instrument in the prospect of the Integrated Pest Management for mate plants.

Identification of a new nucleopolyhedrovirus from naturally-infected Condylorrhiza vestigialis (Guenée) (Lepidoptera: Crambidae) larvae on poplar plantations in South Brazil

A baculovirus was isolated from larvae of Condylorrhiza vestigialis (Guenée) (Lepidoptera: Crambidae), a pest of a forest species known as Poplar (family Salicaceae, genus: Populus) with high economic value. Electron microscopy analysis of the occlusion body obtained from diseased larvae showed polyhedra containing multiple nucleocapsids per envelope. This baculovirus was thus named Condylorrhiza vestigialis multiple nucleopolyhedrovirus (CoveMNPV) and characterized by its DNA restriction endonuclease pattern, polyhedral protein, viral protein synthesis, and infectivity in insect cell lines. Restriction endonuclease profiles of viral DNA digested with five restriction enzymes were obtained and the CoveMNPV genome size was estimated to be 81+/-2.5 kbp. The isolation of the polyhedra (OBs) was done from the crude extract of infected larvae by ultracentrifugation through sucrose gradients. These viral particles were analyzed by denaturing polyacrylamide gel electrophoresis (SDS-PAGE), which showed a strong band with approximately 33 kDa, corresponding to the main protein of the occlusion bodies (polyhedrin). Also, a similar band was observed for CoveMNPV infected Spodoptera frugiperda cells (SF-21 AE) pulse-labeled with [(35)S] methionine and fractionated by SDS-PAGE. Of the four insect cell lines tested for susceptibility to CoveMNPV infection, the SF-21 AE was the most susceptible with occlusion bodies produced in most of the inoculated cells. This is the first record of an NPV from C. vestigialis.

Desenvolvimento in vitro do fungo simbionte de Atta sexdens rubropilosa Forel, 1908 (Hymenoptera: Formicidae) em meio de cultura com diferentes extratos vegetais

The objective of this study was to evaluate the in vitro development of Leucoagaricus gongylophorus, the symbiotic fungus of Atta sexdens rubropilosa in culture medium added with extracts of 24 plant species. The fungus was inoculated in PDA added with vegetables extracts and four controls, in Petri dishes. The dishes were maintained in climate chambers at temperature of 25±1°C in the dark, to period of 42 days. The fungal growth was evaluated weekly by diameter of the colony. The results showed that the symbiotic fungus of these ants has differentiated growth when compared to the extracts provided in the dishes, and the medium supplemented with extracts of arica (Physocalymma scaberrimum), cerejeira (Amburana acreana), cedro (Cedrela fissilis), timbo (Magonia pubescens), leucena (Leucaena leucocephala) e mogno (Swietenia macrophylla) hampered the development, however the extracts of genipapo (Genipa americana), seringueira (Hevea brasiliensis), inga (Inga edulis) e cambara (Vochysia divergens) had better conditions for the development of the symbiotic fungus.