Helio Conte

Cytopathological process by multiple nucleopolyhedrovirus in the testis of Bombyx mori L., 1758 (Lepidoptera: Bombycidae).

A cytopathological methodology was used to analyze infection by Bombyx mori multiple nucleopolyhedrovirus (BmMNPV), a geographic isolate of the family Baculoviridae, in the caterpillar testes of the B. mori. Japanese B. mori strain caterpillar, fifth instar, were inoculated with BmMNPV and their testes were collected and processed for light and transmission electronic microscopy. Epithelial coating cells and interfollicular septa in testes were susceptible to BmMNPV. The first evidence of infection was detected on the 6th day post-inoculation (p.i.) in the external epithelium, and on the 7th day p.i. in the internal epithelium and interfollicular septa. Cytopathological characteristics consisted of hypertrophied nuclei, the formation of virogenic stroma, and the occlusion of virions in polyhedron protein crystals in several stages of development. At the end of the infectious process, cell lysis and release of polyhedra into the extracellular medium occurred. Histopathology revealed early infection foci in the surrounding regions of tracheal insertions, thus underlining the role of the trachea as an infection-spreading organ in insects. This spreading occurs through penetration of the basal lamina, which facilitates entry of the budded virus into the testis. Additionally, an alignment of a partial sequence of the ORF 14 of the BmMNPV geographic isolate with other NPV certified the virus genera.

Midgut morphophysiology in Sitophilus zeamais Motschulsky, 1855 (Coleoptera: Curculionidae).

Sitophilus zeamais is one of the most aggressive pests of stored grains, causing a significant decrease in the nutritional quality of the grains and major losses in economic trade. The foraging capacity of this pest is assigned to its highly efficient digestive system. Investigations on the morphofunctional features of the midgut, which is the most active region of the alimentary canal, are fundamental to understand the feeding habits of this species. In this study, the midgut of adult insects was isolated, processed, and analyzed on light microscopy, scanning and transmission electron microscopy, protein and enzymatic activities determination, including analyses of the starch hydrolysis products. In S. zeamais, the midgut was differentiated into anterior midgut and posterior midgut, and consisted of digestive, regenerative and endocrine cells. The anterior midgut showed high density of regenerative crypts. Cells containing organelles associated with protein synthesis and presence of amylases and lipases indicated that majority of the digestion process occurred in the anterior midgut. The posterior midgut exhibited numerous gastric caeca and peritrophic membrane. Cells with poorly differentiated cytoplasmic into organelles, elongated microvilli, and low enzymatic activities indicated that the posterior midgut was mainly involved in absorption.

Functional Morphology of the Crop and Proventriculus of Sitophilus zeamais (Coleoptera: Curculionidae)

ABSTRACT Sitophilus zeamais Motschulsky, 1855 (Coleoptera: Curculionidae) is one of the most aggressive pests of stored grains, causing significant decrease in the nutritional quality of the grains and major losses in economic trade. The ability of this pest to damage grains and other products is directly related to the morphology of the alimentary canal. Considering the importance of the foregut in the digestion, this study aimed to describe the morphology of the crop and proventriculus in S. zeamais adults. The tissues were isolated, processed, and analyzed by using light microscopy, scanning electron microscopy, and transmission electron microscopy. The crop functioned as a storage organ, and its cells showed functional characteristics related to protein synthesis. High densities of prokaryotic microorganisms and spicules were found in the lumen. The proventriculus exhibited eight chitinous teeth, which were responsible for grinding and filtering food particles. This organ formed the stomodeal valve, which controlled the unidirectional flow of food through the alimentary canal. The alimentary canal is the primary surface of contact between the external environment and an insects internal environment, and knowledge of its morphology is required to better understand the physiology of stored-grain insect pests.

Morpho-functional characterization and esterase patterns of the midgut of Tribolium castaneum Herbst, 1797 (Coleoptera: Tenebrionidae) parasitized by Gregarina cuneata (Apicomplexa: Eugregarinidae)

Tribolium castaneum (Coleoptera: Tenebrionidae) is a common pest of stored grains and byproducts and is normally infected by Gregarina cuneata (Apicomplexa: Eugregarinidae). The life cycle of this parasite includes the sporozoite, trophozoite, gamont, gametocyte, and oocyst stages, which occur between the epithelium and lumen of the hosts midgut. This study aims to describe the morphofunctional alterations in the midgut and determine the esterase patterns in T. castaneum when parasitized by gregarines. To achieve this purpose, midguts of adult insects were isolated, processed, and analysed using light and electron microscopy. We determined total protein content, amylase activity, and the expression and related activities of the esterases by using polyacrylamide gel electrophoresis (PAGE). The midgut of T. castaneum is formed by digestive, regenerative, and endocrine cells. The effects of parasitism on the digestive cells are severe, because the gregarines remain attached to these cells to absorb all the nutrients they need throughout their development. In these cells, the most common alterations observed include expansion and fragmentation of the rough endoplasmic reticulum, development of the smooth endoplasmic reticulum, changes in mitochondrial cristae, cytoplasmic vacuolization, formation of myelin structures, spherites, large intercellular spaces, autophagic vesicles, expansion of the basal labyrinth, and cytoplasmic protrusions. Deposits of glycogen granules were also observed. Amylase activity was reduced in parasitized insects. Regenerative cells were found in disorganized crypts and did not differentiate into new cells, thus, compromising the restoration of the damaged epithelium. Though few morphological alterations were observed in the endocrine cells, results suggest that the synthesis and/or release of hormones might be impaired. Nine esterases (EST-1 to 9) were identified in the midgut of T. castaneum and were expressed in varying levels in response to parasitism. Two additional isoforms of esterases were exclusively identified in the parasitized insects. The results of this study suggest that gregarines alter the morphology and physiology of the midgut. The changes may result in nutritional depletion and the impairment of other physiological processes, such as reproduction and development of the host. Thus, further studies are needed to uncover the possibility of utilizing gregarines as biological controllers of the insect pest population.

Current progress toward vaccine and passive immunization approaches for Strongyloides spp.

Strongyloides stercoralis is a helminth parasite that can infect millions of people worldwide, particularly in tropical, subtropical and temperate regions with poor sanitation. Several aspects of epidemiology, biology and host-parasite interactions of S. stercoralis have been studied, and substantial knowledge has been acquired; however, very few studies on immunotherapeutic control strategies to prevent infection and disease in humans have been conducted. Therefore, this article reviews the current progress and targets toward vaccine and passive immunization approaches for Strongyloides spp.

Bioprospection of immature salivary glands of Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae)

Larval therapy (LT) comprises the application of sterile Calliphoridae larvae for wound debridement, disinfection, and healing in humans and animals. Larval digestion plays a key role in LT, where the salivary glands and gut produce and secrete proteolytic and antimicrobial substances. The objective of this work was to bioprospect the salivary glands of Chrysomya megacephala (Fabricius, 1794) larvae, using ultrastructural, morphological, and histological observations, and the total protein electrophoretic profile. The salivary glands present a deferent duct, originating from the buccal cavity, which bifurcates into efferent ducts that insert through a slight dilatation to a pair of tubular-shaped tissues, united in the region of fat cells. Histologically, the secretion had protein characteristics. Cell cytoplasm presented numerous free ribosomes, autophagic vacuoles, spherical and elongated mitochondria, atypical Golgi complexes, and dilated rough endoplasmic reticulum. In the apical cytoplasm, secretory granules and microvilli secretions demonstrated intense protein synthesis, basal cytoplasm with trachea insertions, and numerous mitochondria. The present work described the ultrastructure and morphology of C. megacephala third instar salivary glands, confirming intense protein synthesis and the molecular weight of soluble proteins.

CHEMICAL CONTROL OF AEDES AEGYPTI: A REVIEW ON EFFECTS ON THE ENVIRONMENT AND HUMAN HEALTH

Aedes mosquitoes are known to be infected by arboviruses causing disease such as dengue, zika fever, and chikunguya fever, and subsequently transmit them, to humans through the bite of infected females. Chemical control is a measure adopted as part of sustainable management and integrated vector control for public health. There are four principal classes of insecticides used for controlling mosquitoes, all being neurotoxic: organochlorides, organophosphates, carbamates, and pyrethroids. The objective of this work was to review reports on the environmental effects of the insecticides most commonly used for controlling Ae. aegypti . This bibliographic study was conducted using articles and books available in the literature with no time restriction. The databases accessed were: Google Scholar, Pub med, SciELO, and ScienceDirect. These insecticides exhibit toxicity to the environment, and may accumulate in food and water and in the body of vertebrates. Resistance to different insecticides is a problem when the mode of control is chemical, because insects survive insecticide application and higher doses are necessary for controlling the vectors. Considering these results, the ideal method would be the newly proposed means of mosquito control using technology related to modern biotechnology .

Histochemical Characterization of the Embryonic Stages in Diatraea saccharalis (Lepidoptera: Crambidae)

Abstract Embryonic development in Diatraea saccharalis F. (Lepidoptera: Crambidae) has been studied by means of whole mounts, histochemical techniques, and light microscopy. Three embryonic stages were identified up to larval eclosion. In the first stage, 0–10 h, intense multiplication of energids occurred, and significant changes in RNA levels were detected in distinct zones before the start of blastulation. During the second stage, 10–24 h, new protein and RNA sites were detected in the egg’s periphery. This was probably because of migration of energids and formation of the blastula. Gastrulation occurs during this stage and is characterized by intense synthetic and mitotic processes. The third stage, 25–168 h, showed ortochromatic and metachromatic regions at the posterior pole and in the yolk, egg periphery, and body of the embryo. Organogenesis closes approximately on the sixth day when the embryo starts feeding on the remaining yolk. Its development is complete by the end of the seventh day on which the larva ecloses.