Comparative study of digestive cells in different midgut regions of stingless bees (Hymenoptera: Apidae: Meliponinae)

Ultrastructure of the digestive cells was analyzed in three midgut regions (anterior, middle and posterior) of stingless bees. Variations occurs in the presence of lipid inclusions in the cells from posterior midgut and presence of double-membraned vesicles associated to microvilli in the anterior midgut. However, basal plasmic membrane infoldings and augmentation of surface area achieved by microvilli ale very similar in all midgut regions. These results not supported the existence of fluid fluxes in the ectoperitrophic space and suggest that digestive cells in stingless bees are polifunctional, that is, there is not midgut region specialized in secretion ol absorption as observed in other insects.

A specialized midgut region in stingless bees (Hymenoptera: Apidae: Meliponinae)

Ultrastructure of the anterior end of the midgut of three Neotropical stingless bees, including an obligate necrophagous species Trigona hypogea, suggests that this midgut region has no role in nutrient absorption because its lumen is reduced by a thick cuticle that lines the outer epithelium of the cardiac valve. A possible involvement of this midgut region in the production of the peritrophic membrane compounds is discussed.

SCANNING ELECTRON-MICROSCOPY OF THE RHODNIUS-NEGLECTUS (HEMIPTERA) LABIAL SALIVARY-GLANDS AFTER STARVATION

Labial salivary glands are found in the majority of insects. They are relatively large, extend back into the thorax, and in Rhodnius, they are cherry red in color due to a pigment derived from traces of hemoglobin absorbed form the gut. In most insects they are acinous shaped, with long excretion channels that present differentiated regions which from salivary reservoirs. The glands may be relatively simple or complexly branched and convoluted. In Rhodnius they are described as being unilobed with no traces of division. The main duct leaves the gland at its anterior extremity. The acini have different kinds of cells but all of them are seen as sources of secretion. Our material has a different shape due to the fact that the animals spent 20 days under starvation conditions. New data are also obtained through treatment with collagenase and HCl. The importance of the study of these glands lies in the fact that it will further understanding of the transmission of Chagas' disease.

Ultrastructure and histochemistry of the mineral concretions in the midgut of bees (Hymenoptera: Apidae)

Mineral concretions in the digestive cells of bees were examined under transmission electron microscope and histochemically. Ultrastructure shows two types of mineral deposits: 1) mineral concretions which are organized in granules with a striking concentrically layered organization of opaque and clear zones and 2) electron dense granules which appear inside small vacuoles (0.4-0.7 mu m). These two structures are present in the apex of the digestive cells of the posterior midgut. Histochemical data reveal that mineral concretions are composed of calcium, iron and uric acid or its salts while calcium determination gives a positive reaction for electron dense granules. Morphological and chemical similarities between the mineral concretions of bees and those described for other insects suggest that they have an important physiological role regulating the composition of the internal environment and to avoid intoxication. Since concretions and granules are structurally distinct, it is suggested that they are functionally different.

Esterase patterns in species of the triatome bug Rhodnius

The aim of the present study was to analyse esterase patterns in three triatomine species of Rhodnius genus. Four loci, Est 1, Est 2, Est 3 and Est 4, were found. The corresponding enzymes were characterized as carboxylesterases (E.C. 3.1.1.1) or cholinesterases (E.C. 3.1.1.8) based on inhibitory experiments, using eserine sulphate, malathion, mercury chloride, p-chloromercuribenzoate (pCMB) and iodoacetamide. Low genetic variability was observed: Est 1, Est 2 and Est 3 were monomorphic in Rhodnius domesticus, Rhodnius robustus and Rhodnius neivai, whereas locus Est 4 was polymorphic in the first two species. The UPGMA analysis based on esterase genotypic frequencies indicated greater similarity between R. domesticus and R. robustus when compared with R. neivai. The present study expands our knowledge about genetic variability among triatomines and accords with the hypothesis that R. domesticus is a species derived from R. robustus.

A comparative study of digestive cells in different midgut regions of stingless bees (Hymenoptera: Apidae: Meliponinae)

Ultrastructure of the digestive cells was analyzed in three midgut regions (anterior, middle and posterior) of stingless bees. Variations occurs in the presence of lipid inclusions in the cells from posterior midgut and presence of double-membraned vesicles associated to microvilli in the anterior midgut. However, basal plasmic membrane infoldings and augmentation of surface area achieved by microvilli are very similar in all midgut regions. These results not supported the existence of fluid fluxes in the ectoperitrophic space and suggest that digestive cells in stingless bees are polifunctional, that is, there is not midgut region specialized in secretion or absorption as observed in other insects.

Protein requirements in larvae and adults of Scaptotrigona postica (Hymenoptera: Apidia, Meliponinae): Midgut proteolytic activity and pollen digestion

The number and degree of digestion of pollen grains in the midgut and rectum, the midgut proteolytic activity and the time of pollen grain passage through the digestive tract in the stingless bee Scaptotrigona postica (Latreille) have been analyzed. The results show similar protein requirements among larvae, nurse bees and queens, as well as between forager bees and old males, but these requirements are higher in individuals from the former groups than in those from the latter. Although protein requirements have been demonstrated to vary according to a bee's activity in the colony, they are similar among bees from different castes or sexes. These changes in feeding behavior are related to the bee's function and to less competition for nourishment among individuals of the colony. It is also noted that pollen grains took between 6 and 28 h to pass through the digestive tract. Pollen grains are irregularly accumulated in the various regions of the midgut, which may reflect functional differentiation throughout the midgut. © 2001 Elsevier B.V.

Targeting Plasmodium ligands on mosquito salivary glands and midgut with a phage display peptide library

Despite vast efforts and expenditures in the past few decades, malaria continues to kill millions of persons every year, and new approaches for disease control are urgently needed. To complete its life cycle in the mosquito, Plasmodium, the causative agent of malaria, has to traverse the epithelia of the midgut and salivary glands. Although strong circumstantial evidence indicates that parasite interactions with the two organs are specific, hardly any information is available about the interacting molecules. By use of a phage display library, we identified a 12-aa peptide-salivary gland and midgut peptide 1 (SM1)-that binds to the distal lobes of the salivary gland and to the luminal side of the midgut epithelium, but not to the midgut surface facing the hemolymph or to ovaries. The coincidence of the tissues with which parasites and the SM1 peptide interact suggested that the parasite and peptide recognize the same surface ligand. In support of this hypothesis, the SM1 peptide strongly inhibited Plasmodium invasion of salivary gland and midgut epithelia. These experiments suggest a new strategy for the genetic manipulation of mosquito vectorial capacity.

Ultrastructural and Cytochemical Aspects of Metamorphosis in the Midgut of Apis mellifera L. (Hymenoptera: Apidae: Apinae)

The midgut of Apis mellifera is remodeled during metamorphosis. The epithelium and, to a lesser extent, the muscular sheath degenerate between the end of the last larval instar and the onset of pupation (prepupa).The larval epithelium is shed to the midgut lumen and digested, while a new epithelium is reconstructed from larval regenerative cells. During pupation, some reorganization still occurs, mainly in brown-eyed pupae. In pharate adult, the midgut wall shows the characteristics of adult, although some cells have pycnotic nuclei. The localization of alkaline and acid phosphatases showed that these enzymes were not involved in the reabsorption of the midgut wall.

Description of the nucleolar activity and karyotype in germinative cell lines of Rhodnius domesticus (Triatominae, Heteroptera)

The purpose of this work was to study the karyotype, spermatogenesis and nucleolar activity at meiosis, in the species Rhodnius domesticus (Heteroptera, Triatominae). The testicular tubules were cytologically prepared by the conventional method of cell crushing and subsequent application of cytogenetic staining techniques with lacto-acetic orcein and silver-ion impregnation. The species under study presented karyotype 2n= 20A+XY, the modal number of the subfamily Triatominae. The chromosomes presented no primary constriction and were therefore characterized as holocentric. It was observed that the sex chromosomes sometimes were located at the periphery, close to the ring formed by autosomes, at first meiotic division. At metaphases II, sex chromosomes were positioned in the center of the autosomal ring, thus evidencing a postreductional behavior. These same chromosomes showed late migration at anaphases and were clearly impregnated with silver-ions, suggesting they bore Nucleolar Organizer Regions. Dispersed nucleolar corpuscles in cytoplasm until telophase II and small dots in spermatids strongly impregnated with silver, could be seen. Thus, it may be inferred that, in triatomines, the nucleolus does not completely disappear but remains in the form of small corpuscles that have a role in cell differentiation.

Estudo biológico e morfométrico dos estádios ninfais de Rhodnius neglectus Lent, 1954 (Hemiptera, Reduviidae)

The following parameters of the biological cycle of the Rhodnius neglectus were determined under a temperature of 28°C and relative humidity of the air varying between 52-94%: a) average period of incubation: 13.67 days; b) rate of the eggs eclosion: 86.18%; c) average period of growth of the 1st, 2nd, 3rd, 4th, 5th instar nymphs: 18.91, 15.19, 21.38, 21.34, 30.43 days, respectively; d) percentage of death in the instar nymphs: 13.82, 18.26, 9.95, 8.78, 15.06% respectively; e) percentage of changes per instar nymphs: 86.18, 81.74, 90.05, 91.22, 84.94% respectively. The morphometric study with the stereoscopic microscope and Leica image analyses system, using the Qwin software, of the five instar nymphs showed that in the five instar nymphs a gradual growth of the head, thorax and abdomen occurs. In all the instars the abdomen is the largest segment. In the four first instars the head is larger than the thorax. In the fifth instar the head and the thorax present equivalent sizes.

Comparative morphology of the midgut of three species of Cephalotes (Hymenoptera, Formicidae, Myrmicinae)

Studies of histological nature, histochemistry and ultra-morphology of the midgut of three species of ants belonging to the genus Cephalotes (atratus, clypeatus and pusillus) were performed. In histology H-E was employed, and in histochemistry techniques for detection of carbohydrates, proteins in general and collagen and calcium fibers were used. The results obtained from this second technique made possible some discussion about the pH of the midgut of these insects. The species are identical in the characters under study. The digestive cells exhibited a secretion that is typical of ants, the apocrine type.

Interaction of Bacillus thuringiensis Cry1 and Vip3A proteins with Spodoptera frugiperda midgut binding sites

Vip3Aa, Vip3Af, Cry1Ab, and Cry1Fa were tested for their toxicities and binding interactions. Vip3A proteins were more toxic than Cry1 proteins. Binding assays showed independent specific binding sites for Cry1 and Vip3A proteins. Cry1Ab and Cry1Fa competed for the same binding sites, whereas Vip3Aa competed for those of Vip3Af. Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Fluazuron-induced morphophysiological changes in the cuticle formation and midgut of Rhipicephalus sanguineus Latreille, 1806 (Acari: Ixodidae) nymphs

The present study demonstrated the effects of the arthropod growth regulator, fluazuron (Acatak®), in the formation of the integument and digestive processes of Rhipicephalus sanguineus nymphs fed on rabbits treated with different doses of this chemical acaricide. For this, three different doses of fluazuron (20, 40, or 80 mg/kg) were applied pour on to the hosts (groups II, III, and IV), as well as distilled water to the control group. On the first day after treatment (24 h), the hosts were artificially infested with R. sanguineus nymphs. After full engorgement (7 days), the nymphs were removed, placed on labeled Petri dishes, and kept in biochemical oxygen demand incubator for 7 days. The engorged nymphs were then taken for morphological, histochemical, and histological analyses. The results showed the occurrence of cytological, morphohistological, and histochemical alterations in the integument and midgut of nymphs from all the different treated groups. These alterations occurred at cuticular level in the subdivisions of the cuticle, related to the size of the digestive cells, amount of accumulated blood elements, and digestive residues, as well as the presence of vacuoles in the cytoplasm of the digestive cells. Thus, this study demonstrated that fluazuron acts on the integument and midgut cells of R. sanguineus nymphs fed on treated rabbits and pointed out the possibility of the use of this chemical - which is more specific, less toxic, and less harmful to the environment and nontarget organisms - in the control of R. sanguineus, at least in the nymphal stage of its biological cycle. © 2012 Springer-Verlag.