Morphology of the Mandibular and Intramandibular Glands of Army Ant Workers of Labidus praedator (Smith 1858) and Labidus coecus (Latreille 1802) (Formicidae: Dorylinae).

Social insects are characterized by having a wide diversity of exocrine glands, with highlights for ants with about 85 glands spreading throughout the body. The mandibular and intramandibular glands are associated with the production of pheromones. The army ants (Dorylinae) play an important role in the structure of the invertebrate community because they are efficient predators and provide suitable conditions for various animals following their invasions in the food search. Labidus coecus (Latreille) is an underground-ameliorating ant and Labidus praedator (Smith) is a generalist surface predator which can deplete invertebrate biomass by up to 75%. This work investigated the morphology of the mandibular and intramandibular glands of L. praedator and L. coecus workers. The glands were analyzed by light microscopy, histochemistry, and scanning electron microscopy. The mandibular and intramandibular glands of the two species were classified as class III glands. The data on the morphology of the mandibular glands has revealed that they have characteristics in common with other subfamilies. The intramandibular glands of the two species of Labidus have similar morphology and chemical composition, which indicates that the components of these glands can have the same function despite their different habits.

Hemocyte morphology of worker subcastes of the leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae).

Hemocytes are cells present in the hemolymph of insects that play a role in combating invasive pathogens, ensuring defense by the immune system in these organisms. While the types of hemocytes are well known in some insect representatives, data on these cells in Hymenoptera are limited to certain bees and wasps, with little information available for ants. Among ants, the genus Atta has environmental and economic importance, forming highly organized colonies consisting of the queen and workers, with the latter subdivided into subcastes: gardeners, waste removers, foragers, and soldiers, which are exposed to different pathogens. This study describes the morphology of hemocytes in the worker subcastes of Atta sexdens rubropilosa. Hemolymph samples from the ant were submitted to light, confocal, and scanning electron microscopy analyses. Five types of hemocytes were identified in the hemolymph of all ant subcastes, including prohemocytes, oenocytoids, spherulocytes, plasmatocytes, and granulocytes. They exhibited nuclei with a predominance of decondensed chromatin. The granulocytes were the most abundant cell type in the subcastes, followed by prohemocytes, plasmatocytes, oenocytoids, and spherulocytes. Phagocytosis assays reveal that plasmatocytes and granulocytes are the main phagocytic cells in all castes evaluated. This study fills an important gap in understanding the immune response in this ant species.

Effect of Induced Pulmonary Arterial Hypertension on Testicular Parameters of Wistar Rats Subjected to Resistance Exercise Training.

Pulmonary arterial hypertension (PAH) is characterized by elevated arterial pressure and vascular resistance. PAH may cause alterations in the microcirculation of several organs, including the kidney, liver, brain, and testes. However, it remains unclear whether monocrotaline-induced PAH exerts detrimental effects on animal testes. Thus, we analyzed the impact of PAH on testicular morphology and function. Additionally, we investigated the effect of resistance exercise training (RT) on testicular parameters in PAH rats. Eight healthy Wistar rats and eight PAH rats were subjected to RT training for 30 days; the other PAH and healthy rats (n = 8/group) did not exercise. PAH rats had lower reproductive organ weight, serum testosterone levels, testicular glucose, and nitric oxide (NO) levels, Leydig cell parameters, tubular morphometry, germ cell counts, and daily sperm production than healthy animals did. The practice of RT attenuated the negative impact of PAH on the relative weights of the testes and epididymides, Leydig cell number, nuclear volume, testicular NO levels, and seminiferous epithelium architecture. Moreover, RT positively influenced testosterone levels in PAH animals. We conclude that PAH exerts deleterious effects on testicular histology and function. However, RT can be beneficial to the PAH-affected testicular parameters.

The fungicide azoxystrobin causes histopathological and cytotoxic changes in the midgut of the honey bee Apis mellifera (Hymenoptera: Apidae).

Apis mellifera is an important bee pollinating native and crop plants but its recent population decline has been linked to the use of pesticides, including fungicides that have been commonly classified as safe for bees. However, many pesticides, in addition to direct mortality cause sublethal effects, including damage to target selective honey bee organs. The midgut is the organ responsible for the digestion and absorption of nutrients and the detoxification of ingested substances, such as pesticides. This study evaluated the histopathological and cytotoxic changes in the midgut of A. mellifera workers caused by the pesticide azoxystrobin. The limit-test was performed, and a 100 µg a.i./bee dose was administered orally and midgut analyzed with light and transmission electron microscopies after 24 h and 48 h of pesticide exposure. The midgut of the control bees has a single layer of digestive cells, with spherical nuclei, nests of regenerative cells, and the lumen coated with the peritrophic matrix. The bees fed on azoxystrobin showed morphological changes, including intense cytoplasm vacuolization and cell fragments released into the gut lumen. The protein detection test showed greater staining intensity in the nests of regenerative cells after 24 h of exposure to azoxystrobin. The occurrence of damage to the midgut in A. mellifera exposed to azoxystrobin indicates that although this fungicide has been classified as low toxicity for bees, it has sublethal effects in the midgut, and effects in other organs should be investigated.

Larval and adult digestive tract of the carrion beetle Oxelytrum discicolle (Brullé, 1840) (Coleoptera: Silphidae).

Oxelytrum discicolle is a necrophagous beetle occurring in Central and South America, with potential use in forensic entomology for investigations in the context of legal medicine. The present work aimed to describe the morphology of the digestive tract of larvae and adults and contributes to the knowledge about the digestion associated with necrophagy. In the three larval instars, the foregut is short and narrow, the midgut is a dilated tube, elongated with a smooth surface, and the hindgut is narrow and long, with small lateral projections and a dilated terminal region. The gut epithelium in the second and third instar larvae is vacuolated in the mid- and hindgut, with high production of apocrine secretions in the midgut lumen. In adults, the foregut is short, with small spines in the cuticular intima that covers the flattened epithelium. The midgut is dilated, with many short gastric caeca with regenerative cells in the blind portion, which differentiate in digestive columnar cells towards the midgut lumen. The hindgut is long and narrow, with an enlarged distal portion with folded epithelium lined by a thin cuticle. The histochemical tests reveal the absence of protein storage granules in the gut epithelium of larvae and adults. The gut characteristics of the carrion beetle O. discicolle are similar to those of other predatory and phytophagous Coleoptera, which may indicate that ancestry may influence the alimentary canal morphology more than the feeding habits.

Differential gene expression by RNA-seq during Alzheimer's disease-like progression in the Drosophila melanogaster model.

Alzheimer's disease (AD) is characterized by progressive, irreversible loss of memory and cognitive function. Drosophila melanogaster and other animal models are used to study several diseases, in order to elucidate unknown mechanisms and develop potential therapies. Molecular studies require biological samples and, for neuropathologies such as AD biopsy of the human brain, are invasive and potentially damaging. The solution is to use animal models, such as D. melanogaster, which is a model organism that can replace mammalian organisms in such studies. In this study, we evaluated the climbing ability and differential gene expression during AD progression due to the amylodoigenic pathway using RNA-seq, and we performed an in silico analysis of a fruit fly AD-like GFP (Green Fluorescent Protein) model with GFP expression in the pan-neural elav driver. A total of 1388 genes were differentially expressed in all analyzed groups. The main pathways related to those Differentially Expressed Genes (DEGs) during aging and AD progression were evaluated using the fly genes and human orthologs, in order to link genomic information to higher-order functional information with gene pathway mapping. We identified pathways present in all analyzed groups, such as metabolic pathways, ribosomal pathways, proteasome pathways and immune system pathways. Some of the genes were validated by qPCR. Knockdown of CG17754 gene by RNAi promoted degeneration in the fly eye, validating these findings in vivo. The identification of similarities in molecular pathways between the transgenic fly AD-like GFP model and mammals related to AD provides new insights into the use of this fly in screening novel anti-AD drugs.

Acute oral exposure to imidacloprid induces apoptosis and autophagy in the midgut of honey bee Apis mellifera workers.

The honey bee Apis mellifera is an important pollinator that increases the yield and quality of crops. In recent years, honey bee populations have declined in some parts of the world, which has been associated with several causes, including pesticides used in agriculture. Neonicotinoids are neurotoxic insecticides widely used in the world with systemic action mode contaminating nectar and pollen that may be consumed by bees. This study evaluated the side effects of imidacloprid in the midgut of A. mellifera after acute oral exposure. Toxicity, histopathology, cytotoxicity, and expression of autophagy-related gene atg1 were evaluated in honey bee workers orally exposed to imidacloprid. The estimated imidacloprid LC50 was 1.44 mg L-1. The midgut epithelium of bees fed on imidacloprid LC50 has the occurrence of cytoplasm vacuoles, enlarged intercellular spaces, disorganization of the striated border, and nuclear pyknosis, with an organ injury index that increases with time exposure. The midgut digestive cells of treated bees have apical protrusions, damaged mitochondria, and autophagosomes that were characterized for content with organelle debris and high expression of atg1. These features indicate the occurrence of high cell death in the midgut of workers exposed to imidacloprid, which may affect the digestibility the physiology of the insect.

Biochemical and morphological characterization of freshwater microalga Tetradesmus obliquus (Chlorophyta: Chlorophyceae).

Tetradesmus is a microalgal genus with biotechnological potential due to its rapid production of biomass, which is plenty in proteins, carbohydrates, lipids, and bioactives. However, its morphology and physiology need to be determined to guide better research to optimize the species cultivation and biocompounds processing. Thus, this study describes the biochemistry and morphology of the strain Tetradesmus obliquus BR003, isolated from a sample of freshwater reservoirs in a Brazilian municipality. In the T. obliquus BR003 dry biomass, we identified 61.6% unsaturated fatty acids, and 3.4% saturated fatty acids. Regarding other compounds, 28.50 ± 1.47 g soluble proteins/100 g, 0.14 ± 0.009 g carotenoids/100 g, 0.76 ± 0.013 g chlorophyll a/100 g, and 0.42 ± 0.015 g chlorophyll b/100 g with a chlorophyll a/b ratio of 1.8 were detected. The main chemical elements found were S, Mg, and P. The cells of BR003 were elliptically curved at the ends and without appendages. Histochemical tests showed carbohydrates distributed in the cytoplasm and pyrenoids, some lipid droplets, and proteins. The cytoplasm is rich in vacuoles, rough endoplasmic reticulum, mitochondria, and chloroplasts. The nucleus has a predominance of decondensed chromatin, and the cell wall has three layers. Chloroplasts have many starch granules and may be associated with a spherical central pyrenoid. To the best of our knowledge, this was the first biochemical description combined with ultrastructural morphological characterization of the strain T. obliquus BR003, grown under standard conditions, to demonstrate specific characteristics of the species.

Handmade face mask can be sterilized without compromising their efficacy / Máscara facial artesanal pode ser esterilizada sem comprometer sua eficácia / La mascarilla facial hecha a mano se puede esterilizar sin comprometer su eficácia

Background and Objectives: COVID-19 caused by SARS-CoV-2 is transmitted by contact, droplets and aerosols. The World Health Organization recommends the mandatory use of surgical masks for health professionals and encourages countries to adhere to their use, en masse, in order to minimize the transmission of the virus. Since the lack of this personal protective equipment is a cause for concern, the objective of this study was to evaluate the characteristics of facial masks regarding (i) the mechanical properties of elongation, rupture stress and resistance to air passage and (ii) the morphometric properties of thickness, fiber diameter and pore distribution after autoclave sterilization. Methods: The masks were made in TNT Spunbonded (100% polypropylene, 60 g/m2 ), autoclaved at 70 ºC for 5 minutes and stored in plastic films at room temperature. Then, the mechanical properties were determined in an Instron universal testing machine and Gurley type automatic densimeter and the morphometric properties in a semi-automatic thickness gauge and scanning electron microscope. Results: It was observed that a cycle of sterilization of facial masks, by moist heat under pressure, increased the blanket's stiffness, did not produce physical damage and did not reduce the blanket's barrier capacity. Conclusion: The sterilization of face masks made of TNT can be considered a strategy to increase safety in their production and use.(AU)

Justificativa e Objetivos: A COVID-19 causada pelo SARS-CoV-2 é transmitida pelo contato, por gotículas e por aerossóis. A Organização Mundial da Saúde recomenda a obrigatoriedade do uso de máscaras cirúrgicas aos profissionais da saúde e incentiva os países a aderirem ao seu uso, em massa, a fim de minimizar a transmissibilidade do vírus. Posto que a falta desse equipamento de proteção individual causa preocupação, o objetivo deste trabalho foi avaliar as características de máscaras faciais quanto (i) às propriedades mecânicas alongamento, tensão de ruptura e resistência a passagem de ar e (ii) às propriedades morfométricas espessura, diâmetro das fibras e distribuição de poros após esterilização em autoclave. Métodos: As máscaras foram confeccionadas em TNT Spunbonded (100% polipropileno, 60 g/m2), autoclavadas a 70 ºC por 5 minutos e armazendadas em filmes plásticos a temperatura ambiente. Na sequência, as propriedades mecânicas foram determinadas em máquina de teste universal Instron e densímetro automático tipo Gurley e as propriedades morfométricas em medidor de espessura semiautomático e microscópio eletrônico de varredura. Resultados: Observou-se que um ciclo de esterilização das máscaras faciais, por calor úmido sob pressão, promoveu o aumento da rigidez da manta, não produziu danos físicos e não diminuiu a capacidade de barreira da manta. Conclusão: A esterilização de máscaras faciais confeccionadas em TNT pode ser considerada uma estratégia para aumentar a segurança na sua produção e uso.(AU)

Antecedentes y objetivos: El COVID-19 causado por el SARS-CoV-2 se transmite por contacto, gotitas y aerosoles. La Organización Mundial de la Salud recomienda el uso obligatorio de mascarillas quirúrgicas para los profesionales de la salud y alienta a los países a adherirse a su uso, en masa, con el fin de minimizar la transmisión del virus. Dado que la falta de este equipo de protección personal es motivo de preocupación, el objetivo de este estudio fue evaluar las características de las máscaras faciales en cuanto a (i) las propiedades mecánicas de elongación, tensión de rotura y resistencia al paso del aire y (ii) la morfometría propiedades de espesor, diámetro de fibra y distribución de poros después de la esterilización en autoclave. Métodos: Las máscaras se fabricaron en TNT Spunbonded (100% polipropileno, 60 g / m2), se esterilizaron en autoclave a 70 ºC durante 5 minutos y se almacenaron en películas plásticas a temperatura ambiente. Posteriormente, se determinaron las propiedades mecánicas en una máquina de ensayo universal Instron y densímetro automático tipo Gurley y las propiedades morfométricas en un medidor de espesor semiautomático y microscópio electrónico escaneando.Resultados: Se observó que un ciclo de esterilización de mascarillas faciales, por calor húmedo a presión, incrementó la rigidez de la manta, no produjo daño físico y no redujo la capacidad barrera de la manta. Conclusión: La esterilización de mascarillas faciales fabricadas con TNT puede considerarse una estrategia para aumentar la seguridad en su producción y uso.(AU)

Behavioral and ultrastructural effects of novaluron on Aedes aegypti larvae.

Chitin synthesis inhibitors (CSI) are supposed to inhibit formation of chitin microfibrils in newly synthesized cuticle during molting process. Conversely, there has been comparatively few data on morphological effects of CSI on non-target insect organs. In this work, the effects of the CSI novaluron on behavior and midgut of A. aegypti were evaluated. Toxicity bioassays revealed that novaluron is toxic to A. aegypti larva with LC50 = 18.57 mg L-1 when exposed in aqueous solution for 24 h. Novaluron treated larvae were less active and spent more time resting compared to the control group. Histopathology showed that midguts of novaluron-treated larvae had cytoplasm vacuolization and damaged brush border. Cytotoxic effects in midguts of treated larvae induced necrosis, autophagy and damage to mitochondria. Despite being chitin synthesis inhibitor, novaluron did not induce alterations in the integument of A. aegypti larvae. Fluorescence microscopy revealed that the number of digestive cells were higher in novaluron-treated larvae than in control, in response to digestive cell apoptosis. The present study highlights the importance of novaluron against A. aegypti larvae by causing injuries to non-target organs, altering behaviors, inducing cell death and inhibiting cell proliferation.

The salivary glands of Brontocoris tabidus (Heteroptera: Pentatomidae): Morphology and secretory cycle.

Brontocoris tabidus (Signoret) (Heteroptera: Pentatomidae) is a zoophytophagous insect used for biological control in agriculture and forest systems because its nymphs and adults feed on insects and plants. The predatory Pentatomidae insert the mouthparts into the prey, releasing saliva to paralysis and kills the insect, as well as digest body parts to be sucked in a preliminary extra-oral digestion. In a short period of time, this insect shows the ability to feed again, suggesting the existence of a constant and abundant secretory cycle in the salivary glands. This study evaluated the morphological, histochemical and ultrastructural changes of the salivary glands of B. tabidus in fed and starved insects. The salivary complex of this predatory bug has a pair of bilobed salivary glands and a pair of tubular accessory salivary glands. The accessory glands have the lumen lined by a thick non-cuticular layer rich in glycoproteins. The secretory cells of the B. tabidus principal salivary glands have constant secretory activity, with each lobe producing different substances. The physiological processes that occur in the salivary gland of B. tabidus indicate that the insect needs to feed constantly, corroborating the potential of this insect to be used in biological control programs.

Spiromesifen induces histopathological and cytotoxic changes in the midgut of the honeybee Apis mellifera (Hymenoptera: Apidae).

The honeybee Apis mellifera is an important pollinator that, similarly to other bees, undergoes colony losses due to several problems, including the use of pesticides in the agriculture. In addition to direct mortality, pesticides cause side-effects in some non-target organs, such as the midgut, which is the main organ for digestion and absorption. Spiromesifen is a pesticide used to control mites and whiteflies, which can be ingested by bees feeding on contaminated floral resources. This study evaluated the histopathological and cytological effects of the ingestion of spiromesifen on the midgut of A. mellifera workers. The bees were exposed per os to the field recommended dose of spiromesifen, and the midgut was analyzed after 24h and 48h of exposure to the pesticide. The midgut has a single layer of digestive cells, with spherical nucleus, nests of regenerative cells and layers of peritrophic matrix in the lumen. Bees treated with spiromesifen presented histological and cytological changes in the midgut, including disorganization of the epithelial architecture, release of cell fragments to the lumen, accumulation of mitochondria in the apical cytoplasm, alteration of the basal labyrinth, changes in the rough endoplasmic reticulum and cell degeneration. The occurrence of damage in the digestive cells of the A. mellifera midgut indicates that spiromesifen does not cause mortality in honeybees, but its side-effects can damage the midgut, which may affect the longevity and behavior of this pollinator.

Arsenic induces dose-dependent structural and ultrastructural pathological remodeling in the heart of Wistar rats.

AIM: Arsenic, an environmental contaminant, represents a public health problem worldwide. Studies have shown its association with molecular mechanisms related to cardiomyocytes redox balance. However, the microstructure and ultrastructure of cardiac tissue, as well as the activity of its antioxidant defenses front of disturbances in the mineral bioavailability induced by arsenic are still scarce. Thus, the aim of this study was to evaluate if arsenic exposure might induce structural and ultrastructural damages in cardiac tissue, including pathological remodeling of the parenchyma and stroma. Moreover, its impact on micromineral distribution and antioxidant enzymes activity in heart tissue was also evaluated. MAIN METHODS: Adult male Wistar rats were divided into three groups that received 0, 1 and 10 mg/L sodium arsenite in drinking water for eight weeks. The hearts were collected and subjected to structural and ultrastructural analysis, mineral microanalysis and antioxidant enzymes quantification. Functional markers of cardiac damages were evaluated using serum samples. KEY FINDINGS: Arsenic exposure induced dose-dependent structural and ultrastructural remodeling of cardiac tissue, with parenchyma loss, increase of stroma components, collagen deposition, and pathological damages such as inflammation, sarcomere disorganization, mitochondria degeneration and myofilament dissociation. Moreover, this metalloid was bioaccumulated in the tissue affecting its micromineral content, which resulted in antioxidant imbalance and increased levels of oxidative stress and cardiac markers. SIGNIFICANCE: Taken together, our findings indicate that the heart is a potential target to arsenic toxicity, and long-term exposure to this metalloid must be avoided, once it might induce several cardiac tissue pathologies.

Morphology and composition of the midgut bacterial community of Scaptocoris castanea Perty, 1830 (Hemiptera: Cydnidae).

The burrower bug Scaptocoris castanea is an important soybean and pasture pest in Brazil, with an underground habit feeding directly on the sap of the roots. Underground habit hinders control and knowledge of the biology and physiology of this pest. This study describes the anatomy, histology, ultrastructure and symbionts of the midgut of S. castanea. The midgut of S. castanea is anatomically divided into five regions (ventricles). Ventricles 1-3 are similar between males and females, with cells specialized in digestion and absorption of nutrients, water transport and homeostasis. Ventricle 4 has squamous epithelium forming crypts and harboring bacteria in the lumen. Ventricle 5 of males is small with cells containing apical microvilli and broad basal folds with many openings for hemocoel, while in females, this region of the midgut is well developed and colonized by intracellular bacteria, characterizing bacteriocytes. The main bacteria are Gammaproteobacteria. The results show sexual dimorphism in ventricle 5 of the midgut of S. castanea, with formation of bacteriocytes in the females, while the other regions are involved in digestive processes in both sexes.

Cytotoxic effects on the midgut, hypopharyngeal, glands and brain of Apis mellifera honey bee workers exposed to chronic concentrations of lambda-cyhalothrin.

The honeybee, Apis mellifera is economically important for its products (honey, wax, and propolis) and for its role in pollination. This insect is threated due to high population losses in both agriculture and beekeeping. Within causes involved in the loss of honeybees is the increased pesticide use on agriculture. Although current testing for the regularization of insecticide use considers its acute toxic effects on pollinators, little is known about the effects of chronic exposure to sublethal concentrations that may persist in the environment. This study investigated the effect of chronic exposure to sublethal concentrations of lambda-cyhalothrin on the midgut, hypopharyngeal glands, and brain of A. mellifera. Honey bees were fed for eight days with LC50/100 insecticide. Subsequently, the midgut, hypopharyngeal glands, and brain were analyzed in light and transmission electron microscopies. The midgut was not affected after exposure, except in the posterior region with cell fragments in the lumen and changes in the mitochondria. The hypopharyngeal glands were severely affected by the insecticide with changes in the rough endoplasmic reticulum and cell death. The brain has extensive gaps in the neuropil as well as in the cellular bodies, especially in the corpora pedunculata. These resembled cellular alterations similar to those seen in death processes. The results of this study indicate that lambda-cyhalothrin is toxic to bees at sublethal concentrations and ingested chronically, causing damage to the midgut, hypopharyngeal glands, and brain, and may affect physiological and behavioral aspects of these insects.

Anatomy, Histology, and Ultrastructure of Salivary Glands of the Burrower Bug, Scaptocoris castanea (Hemiptera: Cydnidae).

The burrower bug Scaptocoris castanea Perty, 1830 (Hemiptera: Cydnidae) is an agricultural pest feeding on roots of several crops. The histology and ultrastructure of the salivary glands of S. castanea were described. The salivary system has a pair of principal salivary glands and a pair of accessory salivary glands. The principal salivary gland is bilobed with anterior and posterior lobes joined by a hilus where an excretory duct occurs. The accessory salivary gland is tubular with a narrow lumen that opens into the hilus near the excretory duct, suggesting that its secretion is stored in the lumen of the principal gland. The cytoplasm of the secretory cells is rich in the rough endoplasmic reticulum, secretory vesicles with different electron densities and mitochondria. At the base of the accessory gland epithelium, there were scattered cells that do not reach the gland lumen, with the cytoplasm rich in the rough endoplasmic reticulum, indicating a role in protein production. Data show that principal and accessory salivary glands of S. castanea produce proteinaceous saliva. This is the first morphological description of the S. castanea salivary system that is similar to other Hemiptera Pentatomomorpha, but with occurrence of basal cells in the accessory salivary gland.

Pyriproxyfen, a juvenile hormone analog, damages midgut cells and interferes with behaviors of Aedes aegypti larvae.

Juvenile hormone analogs (JHA) are known to interfere with growth and biosynthesis of insects with potential for insecticide action. However, there has been comparatively few data on morphological effects of JHA on insect organs. To determine pyriproxyfen effects on Aedes aegypti larvae, we conducted toxicity, behavioral bioassays and assessed ultrastructural effects of pyriproxyfen on midgut cells. A. aegypti larvae were exposed in aqueous solution of pyriproxyfen LC50 concentrations and evaluated for 24 h. This study fulfilled the toxic prevalence of pyriproxyfen to A. aegypti larvae (LC50 = 8.2 mg L-1). Behavioral observations confirmed that pyriproxyfen treatment significantly changes swimming behavior of larvae, limiting its displacement and speed. The pyriproxyfen causes remarkable histopathological and cytotoxic alterations in the midgut of larvae. Histopathological study reveals presence of cytoplasmic vacuolization and damage to brush border of the digestive cells. The main salient lesions of cytotoxic effects are occurrence of cell debris released into the midgut lumen, cytoplasm rich in lipid droplets, autophagosomes, disorganized microvilli and deformed mitochondria. Data suggest that pyriproxyfen can be used to help to control and eradicate this insect vector.

Squamocin induce histological and ultrastructural changes in the midgut cells of Anticarsia gemmatalis (Lepidoptera: Noctuidae).

Annonaceous acetogenins (Annona squamosa Linnaeus) comprises of a series of natural products which are extracted from Annonaceae species, squamocin proved to be highly efficient among those agents. Squamocin is mostly referred as a lethal agent for midgut cells of different insects, with toxic effects when tested against larva of some insects. In present study, LC50 and LC90 of squamocin for A. gemmatalis Hübner (Lepidoptera: Noctuidae) were calculated using probit analysis. Morphological changes in midgut cells were analyzed under light, fluorescence and transmission electron microscopes when larvae were treated with LC50 and LC90 of squamocin for 24, 48 and 72 h. Results revealed that the maximum damage to midgut cells was found under LC90 where it showed digestive cells with enlarged basal labyrinth, highly vacuolated cytoplasm, damaged apical surface, cell protrusions to the gut lumen, autophagy and cell death. The midgut goblet cells showed a strong disorganization of their microvilli. Likewise, in insects treated with squamocin, mitochondria were not marked with Mitotracker fluorescent probe, suggesting some molecular damage in these organelles, which was reinforced by decrease in the respiration rate in these insects. These results demonstrate that squamocin has potential to induce enough morphological changes in midgut through epithelial cell damage in A. gemmatalis.

Male reproductive tract and spermatozoa ultrastructure in the grasshopper Orphulella punctata (De Geer, 1773) (Insecta, Orthoptera, Caelifera).

Identification Orphulellini grasshoppers (Acrididae: Gomphocerinae) species has been difficult due to high polymorphism rate. Orphulella Giglio-Tos, 1894 is a genus with widespread geographical distribution and poor descriptions. Orphulella punctata (De Geer, 1773) has an extensive record of occurrence and available information about the phallic complex, however, there is poor data describing other parts of the male reproductive tract. The objective of this study was characterizes the internal organs of the male reproductive system and spermatozoa of O. punctata. Orphulella punctata testes are of Fountain type, each having only four follicles. Spermatozoa into the seminal vesicle are arranged in bundles with c.a. 2320 µm length, with a nucleus 110 µm long. The spermatozoa are covered by a glycocalyx, the nucleus is cylindrical with condensed chromatin and connected to the flagellum by a dense and lamellar centriole adjunct. The axoneme have 9 + 9 + 2 pattern and present two symmetrical mitochondrial derivatives. A fibrous net and two flat membranous cisternae fill the space between the axoneme and mitochondrial derivatives. This is the first description of the reproductive system of a Gomphocerinae representative.

Dechorionation and Permeabilization of Podisus nigrispinus (Heteroptera: Pentatomidae) Eggs: Limiting Factors for Cryopreservation.

Cryopreservation protocols have been developed for eggs of Diptera, Lepidoptera, and Hymenoptera; however, for Heteroptera, such as Podisus nigrispinus Dallas, 1851 (Pentatomidae), no procedures have been described yet. The objective of this study was to evaluate the processes of dechorionation and permeabilization on the viability of eggs of P. nigrispinus with different embryonic ages. In the laboratory, embryos of 24, 48, 72, and 96 h of age were submitted to sodium hydroxide, sodium hypochlorite, isopropyl, and hexane solutions for dechorionation and permeabilization. The experiment was carried out in a 4 × 8 factorial scheme. Sodium hydroxide affects embryo viability; however, 96-h-old embryos showed higher viability when compared with 24, 48, and 72-h-old embryos. Microscope observations showed that, after the treatments, the chorion of P. nigrispinus eggs was reduced to 5.11 ± 0.30 µm. These solutions for dechorionation and permeabilization of P. nigrispinus eggs together with embryonic age affect the viability of embryos to be cryopreserved.