Leptomonas wallacei shows distinct morphology and surface carbohydrates composition along the intestinal tract of its host Oncopeltus fasciatus (Hemiptera: Lygaeidae) and in axenic culture

Leptomonas wallacei is a monoxenic trypanosomatid that colonizes the digestive tract of the phytophagous hemipteran Oncopeltus fasciatus. This infection was specific and took place exclusively in midgut intestinal ventricles V3 and V4, and in the hindgut. Abundances of parasites in the hindgut were 54% less than those in the hindgut. Parasites in the hindgut were more slender and had a longer flagellum than those from the hindgut, which were rounded, with a shorter flagellum. Moreover, hindgut forms expressed sugar residues on the cell surface, recognized by the lectins from Griffonia simplicifolia-I (alpha-galactose, alpha-N-acetyl-galactosamine) and Helix pomatia (N-acetyl-galactosamine); those sugar residues were not present in protozoa from the midgut. In culture, parasites were morphologically similar to midgut forms, but differed from them because they did not express sugar residues that bind to lectin (beta-galactose(1-3) N-acetyl-galactosamine) from Arachis hypogaea.     

Metamorphic changes in the hindgut of Sarcophaga ruficornis with special reference to the development of rectal pads

In Sarcophaga ruficornis, during normal metamorphosis the larval hindgut epithelium degenerates and the imaginal hindgut epithelium is formed from the proliferation and differentiation of the cells of the posterior imaginal ring. The newly formed hindgut becomes slightly dilated posteriorly to form the rectal pouch. At four places in this pouch, the epithelium becomes thick and forms the primordia of the rectal pads. The cells in the primordia begin to divide and project into the lumen of the pouch and later form the cortex cells of the rectal pads. The cavity of the primordia becomes filled with blood cells and fibrous material, which give rise to the medulla of the rectal pads.     

The invasion of the midgut of the mosquito Culex (Culex) quinquefasciatus Say, 1823 by the helminth Litomosoides chagasfilhoi Moraes Neto, Lanfredi and De Souza, 1997

The Litomosoides chagasfilhoi helminth was studied as a model for microfilaria invasion of the midgut of Culex quinquefasciatus mosquito, vector of Wuchereria bancrofti helminth, causative agent of the human filariasis. Histology and transmission and scanning electron microscopy were utilized to show the topography of mosquito midgut invasion by the helminth. An analysis of midguts dissected at different time points after a blood meal demonstrated that the microfilariae interacted and crossed the peritrophic matrix and the midgut epithelium of C. quinquefasciatus. The microfilariae invaded preferentially the mosquito abdominal midgut and the invasion process occurred between 2 and 3h after the blood feeding. In some cases, microfilariae caused an opening in the midgut that separated the epithelial cells, while in others cases, the worms caused the detachment of cells from the epithelium. Ultimately, L. chagasfilhoi crossing activity appeared to damage the midgut. It was also observed that the microfilariae lost their sheaths during their passage through the fibrous material of the peritrophic matrix, before they reached the midgut epithelium. Since the exsheathment process is necessary for the continuity of larvae development, it seems that the passage through the peritrophic matrix is an important step for the parasite's life cycle. This experimental model revealed details of the interaction process of helminthes within the vector midgut, contributing to the knowledge of factors involved in the vector competence of C. quinquefasciatus as a vector of filariasis.     

Ultrastructural and biochemical characterization of promastigote and cystic forms of Leptomonas wallacei n. sp. isolated from the intestine of its natural host Oncopeltus fasciatus (Hemiptera: Lygaeidae)

Promastigote forms of a trypanosomatid were isolated from the third and fourth ventricles of the midgut and from the hindgut of the phytophagous hemipteran Oncopeltus fasciatus. Some individuals had adhered to its anterior region, close to the flagellar pocket, or to the flagellum up to four rounded aflagellated forms known as straphangers cysts. Scanning electron microscopy revealed that the flagellated forms presented a twisted cell body and a long flagellum, and the cysts, smaller than the parental promastigote, had a nascent flagellum. Transmission electron microscopy showed that promastigotes were typical, while cystic forms were ovoid dense cells devoid of a cyst wall, but presenting a cell coat, a special subpellicular region limited by a membrane unit, and a condensed cytoplasm. The kinetoplast-DNA fibrils appeared as dense spots and the condensed chromatin was arranged in a labyrinthic structure. Desmosome-like structures, observed in the region of adhesion of the precystic forms to the parental promastigote, could explain how cysts remain attached to the mother cell during the encystation process. Release of membranes from the surface of promastigotes and cysts seems to be correlated with the condensation of the cytoplasm during encystment. Morphological and isozyme analyses indicated that this trypanosomatid belongs to the genus Leptomonas. The molecular karyotype of this isolate was compared with that of a strain of Leptomonas oncopelti obtained from Oncopeltus varicolor by contour-clamped homogeneous electric field (CHEF) electrophoresis and revealed similar DNA banding patterns between 2,200-825 Kb, but not in lower bands (825-225 Kb). This suggested that the isolate from O. fasciatus and that from O. varicolor were not identical. Based on our findings we are describing Leptomonas wallacei n. sp. for our isolate from O. fasciatus.     

Ultrastructure of the midgut and hindgut of Derocheilocaris remanei (Crustacea, Mystacocarida)

Ultrastructural features and structure of the midgut and hindgut of Derocheilocaris remanei were studied. The large endodermal midgut is differentiated into an anterior midgut and a posterior midgut separated by a conspicuous constriction. Both circular and longitudinal striated muscle bands surround the midgut, while the hindgut only presents longitudinal muscles. The limit between the midgut and the cuticle-lined hindgut is marked by a rectal valve. In cross-section, the short hindgut is triradiate and has a distinct Y-shaped lumen. The hindgut cuticular lining appears interrupted at the tip of every branch of the Y. Three different cell types are found in the midgut epithelium: basally located undifferentiated cells that give rise to the other two specialized cell types; secretory zymogen-like cells responsible for extracellular digestion and located mainly in the anterior midgut; and vacuolated cells, distributed all along the midgut and appearing to have several functions, including absorption, intracellular digestion, and nutrient transport. A single basic cell type forms the hindgut epithelium. The suggested function for the hindgut is the transport and ejection of waste products.     

Morphological evidence for proliferative regeneration of the Anopheles stephensi midgut epithelium following Plasmodium falciparum ookinete invasion

Ookinetes are motile invasive stages of the malaria parasite that enter the midgut epithelium of the mosquito vector via an intracellular route. Ookinetes often migrate through multiple adjacent midgut epithelial cells, which subsequently undergo apoptosis/necrosis and are extruded from the midgut epithelium into the midgut lumen. Hundreds of ookinetes may simultaneously invade the midgut epithelium, causing destruction of an appreciable proportion of the total number of midgut epithelial cells. However, there is little evidence that ookinete invasion of the midgut epithelium per se is detrimental to the survival of the mosquito vector implying that efficient mechanisms exist to restore the damaged midgut epithelium following malaria parasite infection. Proliferation and differentiation of precursor stem cells could replace the midgut epithelial cells destroyed and lost as a consequence of ookinete invasion. Although the existence of so-called "regenerative" cells within the mosquito midgut epithelium has long been recognized, there has been no previously published evidence for proliferation/differentiation of these putative precursor midgut epithelial cells in mature adult female mosquitoes. In the current study, examination of Giemsa-stained histological sections from Anopheles stephensi mosquito midguts infected with the human malaria parasite Plasmodium falciparum provided morphological evidence that regenerative cells undergo division and subsequent differentiation into normal columnar midgut epithelial cells. Furthermore, the number of these putatively proliferating/differentiating regenerative cells was significantly higher in P. falciparum-infected compared to uninfected mosquitoes, and was positively correlated with both the level of malaria parasite infection and midgut epithelial cell destruction. The loss of invaded midgut epithelial cells associated with intracellular migration by ookinetes, therefore, appears to trigger, and to be compensated by, proliferative regeneration of the mosquito midgut epithelium.     

Morphological and ultrastructural characterization of the alimentary canal in larvae of Streltzoviella insularis (Staudinger) (Lepidoptera: Cossidae)

Streltzoviella insularis (Staudinger) is an important tree‐boring pest, that primarily damages Sophora japonica (Linnaeus) and Ginkgo biloba (Linnaeus), as well as other common species, at great economic cost to the urban landscape construction industry in China. In the present study, the alimentary canal morphology of S. insularis was observed using light microscopy, and its ultrastructure was investigated by scanning and transmission electron microscopy. The foregut of S. insularis can be divided into the pharynx, esophagus, crop, proventriculus, and cardiac valve. The well‐developed crop forms the longest section of the foregut. It is able to store large amounts of food and is lined with a monolayer of epithelial cells. Many sclerotized microspines occur on the surface of the anterior intima and there are dense spines on the posterior intima of the proventriculus. Epithelial cells of the midgut include columnar cells, goblet cells, and regenerative cells, but endocrine cells are absent. The hindgut consists of the pyloric valve, ileum, and rectum. There is no clear distinction between the ileum and colon. The intima surface of the pyloric valve carries many microspines, whereas the intestinal wall of the rectum is thin with well‐developed rectal pads. The rectal epithelial cells form a squamous monolayer. A cryptonephric excretory system is located in the hindgut. There are six spiral Malpighian tubules, in which a cellular layer on a basement membrane encloses a lumen. These results will provide the basis for further studies of the structure and function in S. insularis larvae.     

太白蝎蛉消化道形态学与组织学研究

利用光学显微镜和扫描电子显微镜, 在形态学和组织学水平上研究了太白蝎蛉Panorpa obtusa Cheng成虫消化道的结构。结果表明： 蝎蛉消化道由前肠、中肠、和后肠组成。前肠包括咽喉、食道、和前胃, 但没有嗉囊,其中咽喉可分为骨化的前咽和附着扩肌的后咽（咽喉唧筒）; 前胃壁很厚,内膜上长有许多排列整齐、紧密的棕色胃刺,司过滤、暂时储存和磨碎食物的功能; 前肠末端有6个贲门瓣伸入中肠。中肠较长且膨大,其肠壁细胞由柱状细胞和再生细胞组成; 肠壁细胞外分别为环肌和纵肌,无胃盲囊,也未观察到围食膜。6根棕红色的马氏管位于中、 后肠分界处。后肠分为不对称的“V”字型回肠、环状结肠、以及膨大透明的直肠, 直肠内壁上有6个交替排列的直肠垫。最后简要讨论了蝎蛉消化道的结构与功能,及其在蝎蛉科昆虫分类中的意义。     

胡氏边白蚁消化系统的微细构造

胡氏边白蚁Marginitermes hubbardi(Banks)消化系统可分为前肠、中肠及后肠三大段.前肠包括葡萄状唾腺、口、咽喉、食道、前胃及贲门瓣;从贲门瓣开始到马氏管着生处为止这一段为中肠;后肠则分为葫芦形胃、结肠、直肠和肛门.其消化系统的特点:在前、后肠有几丁内膜、细胞层上还有一层微绒毛;上皮细胞底膜内陷很深,折叠中夹着许多线粒体;中肠围食膜表面有几丁层一直延伸到后肠;后肠前端膨大的葫芦胃中共生了很多种细菌及原生动物,共生的细菌、动物分泌纤维素酶帮助它消化木质纤维.     

Foxh1 and Foxa2 are not required for formation of the midgut and hindgut definitive endoderm

The definitive endoderm forms during gastrulation and is rapidly transformed into the gut tube which is divided along the anterior-posterior axis into the foregut, midgut, and hindgut. Lineage tracing and genetic analysis have examined the origin of the definitive endoderm during gastrulation and demonstrated that the majority of definitive endoderm arises at the anterior end of the primitive streak (APS). Foxh1 and Foxa2 have been shown to play a role in specification of the APS and definitive endoderm. However, prior studies have focused on the role of these factors in specification of foregut definitive endoderm, while their role in the specification of midgut and hindgut definitive endoderm is less understood. Furthermore, previous analyses of these mutants have utilized definitive endoderm markers that are restricted to the anterior endoderm, expressed in extraembryonic endoderm, or present in other germ layers. Here, we characterized the expression of several novel definitive and visceral endoderm markers in Foxh1 and Foxa2 null embryos. In accordance with previous studies, we observed a deficiency of foregut definitive endoderm resulting in incorporation of visceral endoderm into the foregut. Interestingly, this analysis revealed that formation of midgut and hindgut definitive endoderm is unaffected by loss of Foxh1 or Foxa2. This finding represents a significant insight into specification and regionalization of mouse definitive endoderm.     

Pathological changes in Fenneropenaeus indicus experimentally infected with white spot virus and virus morphogenesis

To demonstrate pathological changes due to white spot virus infection in Fenneropenaeus indicus, a batch of hatchery bred quarantined animals was experimentally infected with the virus. Organs such as gills, foregut, mid-gut, hindgut, nerve, eye, heart, ovary and integument were examined by light and electron microscopy. Histopathological analyses revealed changes hitherto not reported in F. indicus such as lesions to the internal folding of gut resulted in syncytial mass sloughed off into lumen, thickening of hepatopancreatic connective tissue with vacuolization of tubules and necrosis of rectal pads in hindgut. Virus replication was seen in the crystalline tract region of the compound eye and eosinophilic granules infiltrated from its base. In the gill arch, dilation and disintegration of median blood vessel was observed. In the nervous tissues, encapsulation and subsequent atrophy of hypertrophied nuclei of the neurosecretory cells were found. Transmission electron microscopy showed viral replication and morphogenesis in cells of infected tissue. De novo formed vesicles covered the capsid forming a bilayered envelop opened at one end inside the virogenic stroma. Circular vesicles containing nuclear material was found fused with the envelop. Subsequent thickening of the envelop resulted in the fully formed virus. In this study, a correlation was observed between the stages of viral multiplication and the corresponding pathological changes in the cells during the WSV infection. Accordingly, gill and foregut tissues were found highly infected during the onset of clinical signs itself, and are proposed to be used as the tissues for routine disease diagnosis.     

Characterization of the alimentary canal of the aphidophagous ladybird,Adalia bipunctata (Coleoptera: Coccinellidae): anatomical and histological approaches

The alimentary canal of the two‐spot ladybird Adalia bipunctata (Linnaeus) (Coleoptera: Coccinellidae) presents the foregut (stomodeum), the midgut (mesenteron) and the hindgut (proctodeum). The shortest region is the foregut and the longest is the midgut. The relative proportions of the main regions were found to be similar for males and females. In the foregut it was possible to distinguish the pharynx, the esophagus and the proventriculus but no crop. The hindgut is composed of the ileum, rectum and rectal canal. Generally the organ width is similar for males and females, but females presented a wider proventriculus. The epithelium of the foregut varied from squamous to simple cuboidal and columnar. In the midgut the epithelium is simple columnar with goblet and regenerative cells. The epithelium of the hindgut varied from simple cuboidal to squamous. Females presented thicker midgut epithelium whereas males presented thicker epithelium in the esophagus. The anatomy of the alimentary canal of A. bipunctata seems to conform to its carnivorous and recent phylogenetic status within the family Coccinellidae.     

Fine structure of the rectal pads in Grylloblatta compodeiformis (walker) (Orthoptera : Grylloblattidae) with reference to fluid transport

The rectal pads of the primitive insect Grylloblatta compodeiformis (Orthoptera : Grylloblattidae) were studied using light and electron microscopy. In this species, the rectal epithelium is thickened to form 6 prominent rectal pads, each of which is composed of tall columnar epithelial cells and laterally placed slender junctional cells, but is devoid of secondary cells. The rectal pads are interconnected by simple rectal epithelium, and are lined by a thin cuticular intima. They are surrounded by an extensive connective tissue space, which contains bundles of delicate connective tissue fibers, neurosecretory axons, and tracheae and tracheoles, which do not penetrate into the pads. The epithelial cells exhibit extensive infoldings of the apical plasma membranes that are closely associated with mitochondria. The lateral membranes are also highly folded around large mitochondria that possess longitudinally oriented cristae. These membrane folds form mitochondrial-scalariform junctional complexes and enclose intercellular channels and spaces. The apical cytoplasm of the epithelial cells contains numerous coated vesicles, dense tubular elements, multivesicular bodies and lysosomes, which suggests receptor-mediated endocytosis of macromolecules. The presence of large whorls of rough endoplasmic reticulum and abundant free ribosomes in the cytoplasm and nuclei with multiple, well-developed nucleoli indicate that the epithelial cells are actively engaged in protein synthesis. The ultrastructural features were examined in relation to their role in fluid transport in a cold habitat.     

The fine structure of the midgut epithelium in a centipede,Scolopendra cingulata (Chilopoda,Scolopendridae), with the special emphasis on epithelial regeneration

Scolopendra cingulata has a tube-shaped digestive system that is divided into three distinct regions: fore-, mid- and hindgut. The midgut is lined with a pseudostratified columnar epithelium which is composed of digestive, secretory and regenerative cells. Hemocytes also appear between the digestive cells of the midgut epithelium. The ultrastructure of three types of epithelial cells and hemocytes of the midgut has been described with the special emphasis on the role of regenerative cells in the protection of midgut epithelium. The process of midgut epithelium regeneration proceeds due to the ability of regenerative cells to proliferate and differentiate according to a circadian rhythm. The regenerative cells serve as unipotent stem cells that divide in an asymmetric manner.Additionally, two types of hemocytes have been distinguished among midgut epithelial cells. They enter the midgut epithelium from the body cavity. Because of the fact that numerous microorganisms occur in the cytoplasm of midgut epithelial cells, we discuss the role of hemocytes in elimination of pathogens from the midgut epithelium. The studies were conducted with the use of transmission electron microscope and immunofluorescent methods.     

Effects of phytohemagglutinin (PHA) on the structure of midgut epithelial cells and localization of its binding sites in western tarnished plant bug, Lygus hesperus Knight

Two histological techniques, bright-field microscopy and immunofluoresecent staining were used to elucidate the lethal effect, target tissues and binding sites of phytohemagglutinin (PHA), a lectin from Phaseolus vulgaris L., on the western tarnished plant bug. Bright-field microscopy showed that the nuclei of the foregut epithelial cells were slightly disrupted and elongated but the lumen of the gut was open. The midgut epithelial cells also showed severe disruption. However, the cells of the first and the third ventriculus were much more sensitive to PHA than those in the second ventriculus. The epithelial cells in these two regions were severely disrupted and swollen toward the lumen, resulting in complete closure of the gut. Most of the cells in these regions contained two nuclei. Also, interestingly, the epithelial cells of the hindgut were drastically disrupted leading to complete closure of the lumen. Immunofluoresecent images from the midgut showed that strong binding occurred on brush-border microvilli of the epithelial cells only within the first and third ventriculi, and some signals within their cytoplasm. Thus, immunofluoresecent studies showed that PHA binds preferentially to the midgut region which demonstrates the most severe effects, and that these cells may endocytose the bound PHA.     

Infection and replication sites of Spiroplasma kunkelii (Class: Mollicutes) in midgut and Malpighian tubules of the leafhopper Dalbulus maidis

Spiroplasma kunkelii distribution and infection mechanisms in the intestines and Malpighian tubules of Dalbulus maidis were investigated by transmission electron microscopy. Spiroplasmas were found between microvilli and in endocytic vesicles of the midgut epithelium. At the basal part, cytoplasmic vesicles contained multiple spiroplasmas with tube-like extensions and spiroplasmas accumulated between the laminae rara and densa of the basal lamina. Tip structures of flask-shaped spiroplasmas pierced the lamina densa that was discontinuous in close proximity to spiroplasmas. Spiroplasmas were found in hemolymph, crossed the basal lamina of Malpighian tubule epithelium and accumulated at high numbers in muscle cells that had cytopathogenic changes. S. kunkelii had perithrochous approximately 8nm diameter structures determined to be fimbriae protruding from the cell surface, and similar structures were adhering to the basal lamina of midgut epithelium and to external lamina of muscle cells. Further, spiroplasmas had pili-like appendages at one or both cell poles and appeared to conjugate. This is the first time that fimbriae and pili have been observed in a mollicutes.     

The larval alimentary canal of the Antarctic insect, Belgica antarctica

On the Antarctica continent the wingless midge, Belgica antarctica (Diptera, Chironomidae) occurs further south than any other insect. The digestive tract of the larval stage of Belgica that inhabits this extreme environment and feeds in detritus of penguin rookeries has been described for the first time. Ingested food passes through a foregut lumen and into a stomodeal valve representing an intussusception of the foregut into the midgut. A sharp discontinuity in microvillar length occurs at an interface separating relatively long microvilli of the stomodeal midgut region, the site where peritrophic membrane originates, from the midgut epithelium lying posterior to this stomodeal region. Although shapes of cells along the length of this non-stomodeal midgut epithelium are similar, the lengths of their microvilli increase over two orders of magnitude from anterior midgut to posterior midgut. Infoldings of the basal membranes also account for a greatly expanded interface between midgut cells and the hemocoel. The epithelial cells of the hindgut seem to be specialized for exchange of water with their environment, with the anterior two-thirds of the hindgut showing highly convoluted luminal membranes and the posterior third having a highly convoluted basal surface. The lumen of the middle third of the hindgut has a dense population of resident bacteria. Regenerative cells are scattered throughout the larval midgut epithelium. These presumably represent stem cells for the adult midgut, while a ring of cells, marked by a discontinuity in nuclear size at the midgut-hindgut interface, presumably represents stem cells for the adult hindgut.     

Induction of actin gene expression in the mosquito midgut by blood ingestion correlates with striking changes of cell shape

Ingestion of a blood meal by the female mosquito Anopheles gambiae (L., Diptera: Culicidae), results in a dramatic distention of the midgut epithelium. Here, we report that these events correlate with a transient increase of actin mRNA and protein abundance. The newly synthesized actin may provide a pool of actin protein needed to remodel epithelial cell cytoarchitecture. We also document changes in midgut epithelial cell morphology. Upon blood ingestion, the columnar cells flatten accompanied by the loss of microvilli on the lumenal side and the unfolding of the labyrinth on the basal side. These changes correlate with the large increase of epithelial surface area needed to accommodate the blood meal. Actin gene expression, actin synthesis and cell morphology all return to the pre-feeding state by 24 h after blood intake.