Stephan Malfitano Carvalho

Side-Effects of Thiamethoxam on the Brain and Midgut of the Africanized Honeybee Apis mellifera (Hymenopptera: Apidae)

The development of agricultural activities coincides with the increased use of pesticides to control pests, which can also be harmful to nontarget insects such as bees. Thus, the goal of this work was assess the toxic effects of thiamethoxam on newly emerged worker bees of Apis mellifera (africanized honeybee—AHB). Initially, we determined that the lethal concentration 50 (LC50) of thiamethoxam was 4.28 ng a.i./μL of diet. To determine the lethal time 50 (LT50), a survival assay was conducted using diets containing sublethal doses of thiamethoxam equal to 1/10 and 1/100 of the LC50. The group of bees exposed to 1/10 of the LC50 had a 41.2% reduction of lifespan. When AHB samples were analyzed by morphological technique we found the presence of condensed cells in the mushroom bodies and optical lobes in exposed honeybees. Through Xylidine Ponceau technique, we found cells which stained more intensely in groups exposed to thiamethoxam. The digestive and regenerative cells of the midgut from exposed bees also showed morphological and histochemical alterations, like cytoplasm vacuolization, increased apocrine secretion and increased cell elimination. Thus, intoxication with a sublethal doses of thiamethoxam can cause impairment in the brain and midgut of AHB and contribute to the honeybee lifespan reduction.

Chemical composition and fumigant effect of essentialoil of Lippia sidoides Cham. and monoterpenes against Tenebrio molitor (L.) (coleoptera: tenebrionidae)

A composicao quimica do oleo essencial de Lippia sidoides obtido por hidrodestilacao foi caracterizada e quantificada por GC/MS, bem como sua atividade inseticida por teste de fumigacao foi avaliada sobre Tenebrio molitor. Alem disso, a toxicidade dos monoterpenos carvacrol, 1,8-cineol e timol, tambem foi avaliada quando esses compostos foram aplicados isoladamente, ou em misturas binarias (1:1), ou terciarias (1:1:1). O oleo essencial de L. sidoides tem como principais constituintes o carvacrol (31,68%), ρ-cimeno (19,58%), 1,8-cineol (9,26%) e ϒ-terpineno (9,21%), em 21 compostos identificados, sendo 92,53% do total. Ambos os compostos possuem atividade inseticida contra T. molitor, seguindo a ordem de toxicidade: carvacrol > 1,8-cineol > oleo essencial de L. sidoides > timol; sendo suas respectivas CL50 a 24 h de 5,53; 5,71; 8,04 e 14,71 µL/L ar. Quando as diferentes misturas de carvacrol, 1,8-cineol e timol foram avaliadas contra T. molitor, verificou-se o efeito sinergico. Para a mistura de carvacrol:1,8-cineol a CL50 foi de 5,34 µL/L ar; carvacrol:timol de 7,67 µL/L ar; 1,8-cineol:timol de 7,51 µL/L ar e carvacrol:1,8-cineol:timol de 6,34 µL/L ar. Principalmente o monoterpeno timol teve efeito sinergico, aumentando a toxicidade de carvacrol e 1,8-cineol quando em misturas binarias, tais como carvacrol:timol e 1,8-cineol:timol.

Effects of sublethal dose of fipronil on neuron metabolic activity of Africanized honeybees.

Fipronil is a neurotoxic insecticide that inhibits the gamma-aminobutyric acid receptor and can affect gustative perception, olfactory learning, and motor activity of the honeybee Apis mellifera. This study determined the lethal dose (LD50) and the lethal concentration (LC50) for Africanized honeybee and evaluated the toxicity of a sublethal dose of fipronil on neuron metabolic activity by way of histochemical analysis using cytochrome oxidase detection in brains from worker bees of different ages. In addition, the present study investigated the recovery mechanism by discontinuing the oral exposure to fipronil. The results showed that mushroom bodies of aged Africanized honeybees are affected by fipronil, which causes changes in metabolism by increasing the respiratory activity of mitochondria. In antennal lobes, the sublethal dose of fipronil did not cause an increase in metabolic activity. The recovery experiments showed that discontinued exposure to a diet contaminated with fipronil did not lead to recovery of neural activity. Our results show that even at very low concentrations, fipronil is harmful to honeybees and can induce several types of injuries to honeybee physiology.

Composição química e toxicidade de óleos essenciais para o pulgão-verde Schizaphis graminum(Rondani, 1852)

Este trabalho teve como objetivo identificar e quantificar os constituintes dos oleos essenciais de Illicum verum, Ageratum conyzoides,Piper hispidinervume Ocotea odorifera, bem como avaliar a toxicidade para o pulgao-verde Schizaphis graminum. A qualificacao dos constituintes foi realizada por meio de um cromatografo gasoso + espectrometro de massas, e a quantificacao, por um cromatografo gasoso + detector de ionizacao de chama, ambos com uma coluna DB5. O metodo de hidrodestilacao promoveu um rendimento (p/p) de 3,81% para I. verum, 0,46% para A. conyzoides, 2,85% para P. hispidinervume 0,68% para O. odorifera. Ja os componentes majoritarios foram: precoceno (87,0%) e (E)-cariofileno (7,1%) para A. conyzoides; (E)-anetol (90,4%), limoneno (2,6%) e metil-chavicol (1,3%) para I. verum; metil-eugenol (81,2%) e safrol (10,6%) para Ocotea odorifera; e safrol (82,5%) e α-terpinoleno (13,4%) para P. hispidinervum. Pelos testes de toxicidade aguda (24 horas) com folhas de sorgo ou papel-filtro contaminados, verificou-se que o oleo de A. conyzoides foi o mais toxico para o pulgao, com CL50de 7,13 e 7,08 µL oleo/cm2 respectivamente, seguido por O. odorifera com CL50 de 11,80 e 103,00 µL oleo/cm2 respectivamente; I. verum de 51,80 µL oleo/cm2 em ambos os substratos; e o menos toxico foi o oleo essencial de P. hispidinervum, com CL50 de 62,50 e 143,00 µL oleo/cm2, respectivamente. Dessa maneira, sugere-se que o uso dos oleos essenciais pode representar uma nova ferramenta em programas de manejo integrado de pragas.

Calli induction in leaf explants of coffee elite genotypes

Three experiments were carried out with the objective of achieving high effectiveness in calli induction from high heterozygosis leaf explants of Coffea arabica through indirect somatic embryogenesis. A randomized-block design in a 2x5 factorial arrangement made up of two media [BOXTEL & BERTHOULY (1996) and TEIXEIRA et al. (2004)] and five C. arabica genotypes were used in the first experiment. In the second experiment the embryogenic calli production potential was evaluated in ten genotypes. Each of them was considered as a treatment. In the third experiment the variations in both 2.4-D (2.5 e 20µM) and 2-iP (2.5 e 20µM) concentrations in TEIXEIRA et al. (2004) medium and secondary media were evaluated. Crops were kept in a growth room under darkness, at 25±2oC. The medium described by TEIXEIRA et al (2004) was found to be superior when compared to that described by BOXTEL & BERTHOULY (1996) in the 2.2 and 7.2 genotypes. An opposite behavior was noticed in 4.2 genotype, that is, BOXTEL & BERTHOULY (1996) had medium superiority. Both 3.0 and 5.0 genotypes had the same behavior in both media studied, which shows that the somatic embryo production depends on the genotype. Calli induction depends on the 2-iP and 2.4 D ratio. The 20.0µM of 2.4-D and 20.0µM of 2-iP combination caused the highest embryogenic calli induction rate.