BackgroundHematophagous insects digest large amounts of host hemoglobin and release heme inside their guts. In Rhodnius prolixus, hemoglobin-derived heme is detoxified by biomineralization, forming hemozoin (Hz). Recently, the involvement of the R. prolixus perimicrovillar membranes in Hz formation was demonstrated.Methodology/Principal FindingsHz formation activity of an α-glucosidase was investigated. Hz formation was inhibited by specific α-glucosidase inhibitors. Moreover, Hz formation was sensitive to inhibition by Diethypyrocarbonate, suggesting a critical role of histidine residues in enzyme activity. Additionally, a polyclonal antibody raised against a phytophagous insect α-glucosidase was able to inhibit Hz formation. The α-glucosidase inhibitors have had no effects when used 10 h after the start of reaction, suggesting that α-glucosidase should act in the nucleation step of Hz formation. Hz formation was seen to be dependent on the substrate-binding site of enzyme, in a way that maltose, an enzyme substrate, blocks such activity. dsRNA, constructed using the sequence of α-glucosidase gene, was injected into R. prolixus females' hemocoel. Gene silencing was accomplished by reduction of both α-glucosidase and Hz formation activities. Insects were fed on plasma or hemin-enriched plasma and gene expression and activity of α-glucosidase were higher in the plasma plus hemin-fed insects. The deduced amino acid sequence of α-glucosidase shows a high similarity to the insect α-glucosidases, with critical histidine and aspartic residues conserved among the enzymes.Conclusions/SignificanceHerein the Hz formation is shown to be associated to an α-glucosidase, the biochemical marker from Hemipteran perimicrovillar membranes. Usually, these enzymes catalyze the hydrolysis of glycosidic bond. The results strongly suggest that α-glucosidase is responsible for Hz nucleation in the R. prolixus midgut, indicating that the plasticity of this enzyme may play an important role in conferring fitness to hemipteran hematophagy, for instance.

Oligonychus ilicis (McGregor) (Acari) (Tetranychidae) é uma das principais pragas de Coffea canephora Pierre & Froehner. Neste trabalho, avaliou-se o efeito de diferentes temperaturas no desenvolvimento desse ácaro-vermelho em folhas de C. canephora, em laboratório. Confinaram-se 80 fêmeas em 40 arenas, constituídas de disco foliar de 4 cm, para oviposição nas temperaturas de 21, 24, 27, 30 e 33 ºC. Foram selecionados ao acaso pelo menos dois ovos para avaliar o desenvolvimento embrionário. Após a eclosão da larva, foram realizadas avaliações a cada 12 horas para obtenção da duração e sobrevivência larval. Para avaliar a longevidade foi transferido um ácaro macho da criação para as arenas com uma fêmea para acasalamento. O limite térmico de desenvolvimento inferior e a constante térmica foram determinados para a duração do desenvolvimento de ovo a adulto. O tempo de desenvolvimento das fases imaturas diminuiu com o aumento da temperatura. As fases de ovo, larva, protocrisálida, protoninfa, deutocrisálida, deutoninfa e teleiocrisálida variaram, respectivamente, de 10,4 a 4,3; 2,4 a 1,2; 2,0 a 1,0; 2,3 a 1,2; 1,9 a 1,0; 3,0 a 1,5 e 2,3 a 1,0 dias. O aumento da temperatura afetou o período de desenvolvimento, reduzindo a duração de ovo-adulto e a longevidade. O limite térmico inferior foi de 9,0 ºC para o período de ovo-adulto e constante térmica de 256,4 graus-dias. Em condições de laboratório, a faixa de temperatura que favorece o desenvolvimento do ácaro em C. canephora foi de 24 a 30 ºC. O limite térmico inferior não é limitante para ocorrência de O. ilicis em áreas cultivadas com C. canephora no estado do Espírito Santo.

Hemozoin (Hz), a heme crystal first known as a malaria pigment, reduces heme toxicity to hematophagous organisms such as protozoans, worms, and insects. The mechanism of Hz synthesis remains poorly understood, but studies on the blood-sucking insect Rhodnius prolixus indicate the involvement of an α-glucosidase enzyme. The objective of this study was to test the hypothesis that α-glucosidases from other organisms also have the ability to form Hz in vitro. This hypothesis was tested using protein extracts from non-hematophagous insects and non-insect organisms. Our results indicate that protein extracts from several sources have the potential to crystallize heme in vitro. This activity is likely associated with α-glucosidases. Thus, these enzymes seem to have the secondary capacity of detoxifying heme even in nonhematophagous organisms. This latent function may represent a crucial pre-adaptive evolutionary step in the adoption of hematophagy in hemoparasites.

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