Clóvis Andrade Neves

Ultrastructure of the Digestive Cells in the Midgut of the Predator Brontocoris tabidus (Heteroptera: Pentatomidae) after Different Feeding Periods on Prey and Plants

ABSTRACT Brontocoris tabidus (Signoret) (Heteroptera: Pentatomidae) is an obligate zoophytophagous predator because its population can be maintained in the laboratory when fed on both prey and plants. We evaluated ultrastructural changes in the midgut digestive cells of adult B. tabidus, subjected to different treatments (starvation or feeding on plant material and prey) for different periods. Their midguts were dissected, divided into anterior, medium and posterior sections, processed, and analyzed with light and transmission electron microscopy. The anterior region of the midgut of B. tabidus, starved or fed on eucalyptus leaves, contained no glycogen. B. tabidus fed on plant material showed multivesicular bodies in this region, and spherocrystals after 6 h of feeding on prey. The microvilli of the medium midgut were longer than those of the anterior and posterior midgut. The posterior midgut differed from the other two regions by an abundance of mitochondria, rough endoplasmatic reticulum and double membrane vesicles in the apical region, 6 h after feeding. The ultrastructural features of the digestive cells in the anterior, medium and posterior regions of the midgut suggest that they play a role in digestive enzyme synthesis, ion and nutrient absorption, and storage and excretion of substances.

Notes on Midgut Ultrastructure of Cimex hemipterus (Hemiptera: Cimicidae)

ABSTRACT This work studied the ultrastructure of the midgut cells of Cimex hemipterus Fabricius (Hemiptera: Cimicidae). The midgut of adult insects was analyzed on different days after a bloodmeal, and three anatomical regions with different digestive functions were apparent. In the anterior midgut, the digestive cells had many spherocrystals, lipid inclusions, and glycogen deposits, suggesting a role in water absorption, ion regulation, digestion, and storage of lipids and sugars. The digestive cells in the middle midgut contained secretory granules in the apical cytoplasm, lysosomes, and large amounts of rough endoplasmic reticulum, suggesting that this midgut region was active in digestive processes. The posterior midgut contained digestive cells with secretory vesicles, lysosomes, rough endoplasmic reticulum, and spherocrystals, suggesting digestion and ion/water absorption. Also, there was strong evidence that the posterior midgut may be the major site of nutrient absorption. The hematophagous heteropteran groups share many of these blood digestion mechanisms.

Bark extract of Bathysa cuspidata attenuates extra-pulmonary acute lung injury induced by paraquat and reduces mortality in rats

This study investigated the effect of the bark extract of Bathysa cuspidata on paraquat (PQ)‐induced extra‐pulmonary acute lung injury (ALI) and mortality in rats. ALI was induced with a single dose of PQ (30 mg/kg, i.p.), and animals were treated with B. cuspidata extract (200 and 400 mg/kg). Analyses were conducted of survival, cell migration, lung oedema, malondialdehyde, proteins carbonyls, catalase, superoxide dismutase, histopathology and the stereology of lung tissue. Rats exposed to PQ and treated with 200 and 400 mg of the extract presented lower mortality (20% and 30%), compared with PQ alone group (50%). Furthermore, lung oedema, septal thickening, alveolar collapse, haemorrhage, cell migration, malondialdehyde and proteins carbonyl levels decreased, and catalase and superoxide dismutase activity were maintained. These results show that the bark extract of B. cuspidata reduced PQ‐induced extra‐pulmonary ALI and mortality in rats and suggest that these effects may be associated with the inhibition of oxidative damage.

Trypanosoma cruzi infection induces morphological reorganization of the myocardium parenchyma and stroma, and modifies the mechanical properties of atrial and ventricular cardiomyocytes in rats

BACKGROUND This study investigates morphofunctional adaptations of the heart stroma and parenchyma in rats that are chronically infected with Trypanosoma cruzi. METHODS Four-month-old male Wistar rats were randomized into control (n=14) and infected (n=14) groups. Infected animals were inoculated with T. cruzi Y strain. After 9 weeks, the animals were euthanized, and the right atrium (RA) and left ventricle (LV) were removed for biochemical, stereological, and cardiomyocyte mechanical analyses. RESULTS Infected animals presented cardiomyocyte atrophy and myocardial fibrosis. For these animals, the total volume, length, surface area, and cross-sectional area of cardiomyocytes were significantly reduced, and the total interstitial and collagen volumes were significantly increased in the RA and LV compared to the controls. The total volume and length of blood vessels were significantly increased in the LV, and the total blood vessel surface area was significantly higher in the RA of infected animals. RA and LV cardiomyocytes from infected animals exhibited a significant reduction in cell shortening (43.02% and 24.98%, respectively), prolongation of the time to the peak of contraction (17.09%) and the time to half relaxation (23.68%) compared to non-infected animals. Lipid hydroperoxides, but not mineral concentrations, were significantly increased in the RA and LV from infected animals, showing an inverse correlation with cell shortening. CONCLUSIONS T. cruzi infection induces global structural remodeling of the RA and LV in rats. This remodeling coexists with cardiomyocyte contractility dysfunction, which is possibly related to the abnormal organization of the myocardial stroma and increased cellular lipid peroxidation.

Aedes aegypti midgut remodeling during metamorphosis

The Aedes aegypti midgut is restructured during metamorphosis; its epithelium is renewed by replacing the digestive and endocrine cells through stem or regenerative cell differentiation. Shortly after pupation (white pupae) begins, the larval digestive cells are histolized and show signs of degeneration, such as autophagic vacuoles and disintegrating microvilli. Simultaneously, differentiating cells derived from larval stem cells form an electron-dense layer that is visible 24h after pupation begins. Forty-eight hours after pupation onset, the differentiating cells yield an electron-lucent cytoplasm rich in microvilli and organelles. Dividing stem cells were observed in the fourth instar larvae and during the first 24h of pupation, which suggests that stem cells proliferate at the end of the larval period and during pupation. This study discusses various aspects of the changes during midgut remodeling for pupating A. aegypti.

Ultrastructural characterization of the spermatozoa of Aethalion reticulatum Linnaeus 1767 (Hemiptera: Auchenorrhyncha: Aethalionidae).

The Hemiptera order is currently divided into four suborders. Among them the Auchenorrhyncha suborder is considered to be paraphyletic. Morphology of insect spermatozoa has provided promising characteristics that can be used for phylogenetic inference. In this study, Aethalion reticulatum (Aethalionidae) spermatozoa were examined by light and electron microscopy. The head of the spermatozoa is composed of an acrosome and a nucleus. The nucleus is linear and filled with compact chromatin but has electron-lucid spaces. The centriole adjunct initiates parallel to the nucleus and terminates in the region anterior to the mitochondrial derivatives. Flagella consist of an axoneme, two mitochondrial derivatives and two accessory bodies and the axoneme has the typical 9+9+2 microtubule pattern. The mitochondrial derivatives are symmetric. The accessory bodies are long and are shaped like a half moon when viewed from a cross-section. The presence of accessory bodies differs from other species of Cicadomorpha previously studied. Spermatozoa morphology of other Auchenorrhyncha families can reveal synapomorphies and contribute to systematics of the suborder.

Effects of Trypanosoma cruzi infection on myocardial morphology, single cardiomyocyte contractile function and exercise tolerance in rats

The aim of this study was to investigate the effects of Trypanosoma cruzi (T. cruzi) infection on myocardial morphology, single cardiomyocyte contractile function and exercise tolerance in rats. Adult Wistar rats were randomized into control (n = 14) and infected (n = 14) groups. Infected animals were inoculated with T. cruzi Y strain (300,000 trypomastigotes/50 g body weight). After 9 weeks, the animals were subjected to a treadmill running protocol. Then, the right atrium (RA) and left ventricle (LV) were removed for morphological and cell contractile evaluation. The infected animals exhibited a significant reduction in distance travelled, total time to fatigue and workload. In addition, these animals had hypertrophy, increased myocardial cellularity, and an increase in the proportion of collagen and blood vessels. RA and LV myocytes from infected animals showed marked contractile dysfunction under basal conditions and a reduced contractile response to β‐adrenergic stimulation. The workload of infected animals was correlated closely with the amplitude of cell shortening of RA and LV myocytes. T. cruzi infection influenced the myocardial morphology and the mechanical properties of RA and LV single myocytes negatively and reduced exercise tolerance. Single cardiomyocyte contractile dysfunction could constitute an additional mechanism of cardiac impairment and reduced exercise tolerance in this infection.

Flaxseed, olive and fish oil influence plasmatic lipids, lymphocyte migration and morphometry of the intestinal of Wistar rats.

PURPOSE Evaluate the effect of flaxseed, olive and fish oil on the lipid profile, preservation of villosities and lymphocyte migration in the intestinal mucosa of Wistar rats. METHODS Thirty Wistar male rats were divided into four groups, which received the AIN-93M diet, with changes only to their lipid source: flaxseed, olive, fish, and soy oil (control group). The serum was separated for the biochemical parameter analysis. A histological evaluation was performed in the ileal portion. RESULTS The group which was fed fish oil presented lower values when compared to the other treatments for Total Cholesterol, High-density Lipoprotein Cholesterol and Triacylglycerol (p<0.05). The animals treated with fish and olive oils presented better intestinal villosities preservation. Less deposition of lymphocytes was observed in the flaxseed group (p<0.001). CONCLUSIONS This study demonstrated that flaxseed, olive and fish oils present different responses than soy oil for the intestinal mucosa preservation and lymphocyte proliferation in Wistar rats.

Analysis of the morphology and distribution of argentaffin, argyrophil and insulin-immunoreactive endocrine cells in the small intestine of the adult opossum Didelphis aurita (Wied-Neuwied, 1826)

The aim of this study was to identify and quantify the argyrophil, argentaffin and insulin-immunoreactive cells (IIC) in the small intestine of the opossum Didelphis aurita. Seven adult male specimens of opossums were investigated. The animals were captured, and their blood insulin levels were determined. After euthanasia, fragments of the small intestine were processed for light microscopy and transmission electron microscopy, and submitted to histochemistry and immunohistochemistry for identification of argyrophil and argentaffin endocrine cells, and IIC. Argyrophil and argentaffin cells were identified in the intestinal villi and Liberkühn crypts, whereas IIC were present exclusively in the crypts. Ultrastructure of the IIC revealed cytoplasmic granules of different sizes and electron densities. The numbers of IIC per mm(2) in the duodenum and jejunum were higher than in the ileum (p<0.05). The animals had low levels of blood insulin (2.8 ± 0.78 μIU/ml). There was no correlation between insulin levels and the number of IIC in the small intestine. The IIC presented secretory granules, elongated and variable morphology. It is believed that insulin secretion by the IIC may influence the proliferation of cells in the Liberkühn crypts, and local glucose homeostasis, primarily in animals with low serum insulin levels, such as the opossum.

Effects of atrazine and picloram on grass carp: acute toxicity and histological assessment

The aim of this study was to evaluate the acute toxicity of atrazine and pícloram separately to grass carp (Ctenopharyngodon idella). Firstly, fingerlings were exposed to nominal concentrations of these herbicides to determine the lethal concentration (LC50) (96 h). After this, the animals were treated with sub-acute concentrations of the herbicides to measure the effects on gill histology. The LC50 (96 h) of the atrazine and picloram were, respectively, 37 mg L−1 and 4.4 mg L−1. Four types of alterations were found in gills exposed to atrazine, which were epithelial lifting, partial cell proliferation, lamellar fusion, and aneurysm. Nominal concentrations of picloram caused epithelial lifting, partial cell proliferation, and lamellar fusion.