Description of the larval and adult hindgut tract of the common spider crab Maja brachydactyla Balss, 1922 (Brachyura, Decapoda, Malacostraca).

Arthropods are the most diversified animals on Earth. The morphology of the digestive system has been widely studied in insects; however, crustaceans have received comparatively little attention. This study describes the hindgut tract of the common spider crab Maja brachydactyla Balss, 1922, in larvae and adults using dissection, light and electron microscopical analyses. The hindgut tract maintains a similar general shape in larvae and adults. Major differences among stages are found in the morphology of epithelial cells and microspines, the thickness of the cuticle and connective-like tissue, and the presence of rosette glands (only in adults). Here, we provide the description of the sub-cellular structure of the folds, epithelium (conformed by tendon cells), musculature, and microspines of the hindgut of larvae and adults of M. brachydactyla. The morphological features of the hindgut of M. brachydactyla are compared with those of other arthropods (Insecta, Myriapoda and Arachnida). Our results suggest that the morphology of the hindgut is associated mainly with transport of faeces. In adults, the hindgut may also exert an osmoregulatory function, as described in other arthropods. At difference from holometabolous insets, the hindgut of M. brachydactyla (Decapoda) does not undergo a true metamorphic change during development, but major changes observed between larval and adult stages might respond to the different body size between life stages.

Comparative Features of the Upper Alimentary Tract in the Domestic Fowl (Gallus gallus domesticus) and Kestrel (Falco tinnunculus): A Morphological, Histochemical, and Scanning Electron Microscopic Study.

The avian alimentary tract has evolved into different histologic structures to accommodate the physical and chemical features of several food types and flight requirements. We compared the esophagus, proventriculus, and gizzard of the domestic fowl, Gallus gallus domesticus (GGD) and kestrels, Falco tinnunculus (FT) using immunohistochemistry and scanning electron microscopy with various stains and lectins [Dolichos biflorus agglutinin (DBA) and Ricinus communis agglutinin I (RCA120)], and α-smooth muscle actin (α-SMA). The esophagus of GGD demonstrated thickened epithelium, muscularis mucosae, and inner circular longitudinal tunica muscularis layers; moderate outer longitudinal tunica muscularis layers; and a true crop. In contrast, the esophagus of FT showed a thin epithelium, no muscularis mucosae, moderate inner longitudinal and thick outer circular tunica muscularis layers, and no true crop. In the proventriculus, the nature of the secretion in GGD was neutral, but that of FT was acidic and neutral. In the gizzard, the muscle coat of GGD by α-SMA had no muscularis mucosae, unlike FT, which had muscularis mucosae. In summary, there are many histologic differences between GGD and FT to meet their different physiologic needs, such as feeding.

Acting target of toosendanin locates in the midgut epithelium cells of Mythimna separate Walker larvae (lepidoptera: Noctuidae).

Toosendanin (TSN), which is extracted from the root bark of Melia toosendan Siebold and Zuccarini, has multiple modes of action against insects. Especially, this compound has a potent stomach poisoning activity against several lepidoptera pests. In this paper, the signs of toxicity, digestive enzymes activity, the histopathological changes and immuno-electron microscopic localization of TSN in the midgut epithelium of Mythimna separate Walker larvae were investigated for better understanding its action mechanism against insects. The bioassay results indicated that TSN has strong stomach poisoning against the fifth-instar larvae of M. separata (LC50 = 252.23 µg/mL). The typical poisoned symptom were regurgitation and paralysis. Activities of digestive enzymes had no obvious changes after treatment with LC80 dose of TSN. The midgut epithelial cells of insect were damaged by TSN, showing the degeneration of microvilli, hyperplasia of smooth endoplasmic reticulum and condensation of chromatin. Immunohistochemical analysis revealed that the gold particles existed on the microvilli of columnar cells and goblet cells, and gradually accumulated with the exacerbation of poisoning symptoms, showing that TSN targets on the microvilli of the midgutcells. Therefore, TSN acts on digestive system and locates in the microvilli of midgutcells of M. separata.

Structure and function of the digestive system in molluscs.

The phylum Mollusca is one of the largest and more diversified among metazoan phyla, comprising many thousand species living in ocean, freshwater and terrestrial ecosystems. Mollusc-feeding biology is highly diverse, including omnivorous grazers, herbivores, carnivorous scavengers and predators, and even some parasitic species. Consequently, their digestive system presents many adaptive variations. The digestive tract starting in the mouth consists of the buccal cavity, oesophagus, stomach and intestine ending in the anus. Several types of glands are associated, namely, oral and salivary glands, oesophageal glands, digestive gland and, in some cases, anal glands. The digestive gland is the largest and more important for digestion and nutrient absorption. The digestive system of each of the eight extant molluscan classes is reviewed, highlighting the most recent data available on histological, ultrastructural and functional aspects of tissues and cells involved in nutrient absorption, intracellular and extracellular digestion, with emphasis on glandular tissues.

The amazing complexity of insect midgut cells: types, peculiarities, and functions.

The insect midgut epithelium represents an interface between the internal and the external environment and it is the almost unique epithelial tissue by which these arthropods acquire nutrients. This epithelium is indeed able to produce digestive enzymes and to support vectorial transport of small organic nutrients, ions, and water. Moreover, it plays a key role in the defense against pathogenic microorganisms and in shaping gut microbiota. Another important midgut function is the ability to produce signaling molecules that regulate its own physiology and the activity of other organs. The two main mature cell types present in the midgut of all insects, i.e., columnar and endocrine cells, are responsible for these functions. In addition, stem cells, located at the base of the midgut epithelium, ensure the growth and renewal of the midgut during development and after injury. In insects belonging to specific orders, midgut physiology is deeply conditioned by the presence of unique cell types, i.e., goblet and copper cells, which confer peculiar features to this organ. This review reports current knowledge on the cells that form the insect midgut epithelium, focusing attention on their morphological and functional features. Notwithstanding the apparent structural simplicity of this organ, the properties of the cells make the midgut a key player in insect development and homeostasis.

The digestive system of xenacoelomorphs.

Interest in the study of Xenacoelomorpha has recently been revived due to realization of its key phylogenetic position as the putative sister group of the remaining Bilateria. Phylogenomic studies have attracted the attention of researchers interested in the evolution of animals and the origin of novelties. However, it is clear that a proper understanding of novelties can only be gained in the context of thorough descriptions of the anatomy of the different members of this phylum. A considerable literature, based mainly on conventional histological techniques, describes different aspects of xenacoelomorphs' tissue architecture. However, the focus has been somewhat uneven; some tissues, such as the neuro-muscular system, are relatively well described in most groups, whereas others, including the digestive system, are only poorly understood. Our lack of knowledge of the xenacoelomorph digestive system is exacerbated by the assumption that, at least in Acoela, which possess a syncytial gut, the digestive system is a derived and specialized tissue with little bearing on what is observed in other bilaterian animals. Here, we try to remedy this lack of attention by revisiting the different studies of the xenacoelomorph digestive system, and we discuss the diversity present in the light of new evolutionary knowledge.

The mechanism of damage by trace amounts of acetamiprid to the midgut of the silkworm, Bombyx mori.

Acetamiprid is widely used for agricultural pest control. However, it remains poorly understood whether the environmental residues of acetamiprid have the potential effects on economic insect. In this study, we evaluated the effects of acetamiprid on silkworm growth and development. The exposure to trace amounts of acetamiprid significantly decreased body weight, viability, and spinning ability. In addition, the activity of trypsin in the midgut was decreased after exposure. DGE and KEGG pathway enrichment analysis revealed that the significantly differentially expressed genes were mainly involved in nutrient metabolism, stress responses, and inflammation pathways. These results, in combination with hematoxylin-eosin staining and transmission electron microscopy, indicated that acetamiprid could cause oxidative damage to midgut, lead to inflammatory responses, and affect the activities of midgut digestive enzymes, thus resulting in abnormal growth and development. Our findings greatly contributed to the evaluation of the effects of acetamiprid residues on other nontarget beneficial insect.

Post-embryonic changes in the hindgut of honeybee Apis mellifera workers: Morphology, cuticle deposition, apoptosis, and cell proliferation.

In insects, the hindgut is a homeostatic region of the digestive tract, divided into pylorus, ileum, and rectum, that reabsorbs water, ions, and small molecules produced during hemolymph filtration. The hindgut anatomy in bee larvae is different from that of adult workers. This study reports the morphological changes and cellular events that occur in the hindgut during the metamorphosis of the honeybee Apis mellifera. We describe the occurrence of autophagosomes and the ultrastructure of the epithelial cells and cuticle, suggesting that cuticular degradation begins in prepupae, with the cuticle being reabsorbed and recycled by autophagosomes in white- and pink-eyed pupae, followed by the deposition of new cuticle in light-brown-eyed pupae. In L5S larvae and prepupae, the hindgut undergoes cell proliferation in the anterior and posterior ends. In the pupae, the pylorus, ileum, and rectum regions are differentiated, and cell proliferation ceases in dark-brown-eyed pupae. Apoptosis occurs in the hindgut from the L5S larval to the pink-eyed pupal stage. In light-brown- and dark-brown-eyed pupae, the ileum epithelium changes from pseudostratified to simple only after the production of the basal lamina, whereas the rectal epithelium is always flattened. In black-eyed pupae, ileum epithelial cells have large vacuoles and subcuticular spaces, while in adult forager workers these cells have long invaginations in the cell apex and many mitochondria, indicating a role in the transport of compounds. Our findings show that hindgut morphogenesis is a dynamic process, with tissue remodeling and cellular events taking place for the formation of different regions of the organ, the reconstruction of a new cuticle, and the remodeling of visceral muscles.

Digestive gland inclusion bodies in queen conch (Lobatus gigas) are non-parasitic.

Unusual inclusion bodies occur within the epithelial cells of the digestive gland of queen conch, Lobatus gigas, and have previously been described as apicomplexan parasites. The aim of this study was to investigate the parasitic features of these inclusion bodies in queen conch. L. gigas from St. Kitts (Caribbean Sea) consistently (100% of n = 61) showed large numbers of ovoid to tri-bulbous dark brown inclusion bodies (15 × 30 µm) within vacuolar cells. Histochemical stains demonstrated iron, melanin, and glycoprotein and/or mucopolysaccharide within the inclusion bodies. Microscopic features indicative of a host response to injury were lacking in every case, as were consistent morphological forms to indicate distinct parasitic stages. Transmission electron microscopy failed to reveal cellular organelles of parasitic organisms and DNA extractions of purified inclusion bodies did not yield sufficient concentrations for successful PCR amplification. Scanning electron microscopy with energy dispersive X-ray analysis revealed a number of elements, particularly iron, within the inclusion bodies. We conclude that the inclusion bodies are not an infectious agent, and hypothesize that they represent a storage form for iron, and potentially other elements, within a protein matrix. Similar structures have been described in the digestive glands of other invertebrates, including prosobranchs.

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.

Midgut morphological changes and autophagy during metamorphosis in sand flies.

During metamorphosis, holometabolous insects undergo significant remodeling of their midgut and become able to cope with changes in dietary requirements between larval and adult stages. At this stage, insects must be able to manage and recycle available food resources in order to develop fully into adults, especially when no nutrients are acquired from the environment. Autophagy has been previously suggested to play a crucial role during metamorphosis of the mosquito. Here, we investigate the overall morphological changes of the midgut of the sand fly during metamorphosis and assess the expression profiles of the autophagy-related genes ATG1, ATG6, and ATG8, which are associated with various steps of the autophagic process. Morphological changes in the midgut start during the fourth larval instar, with epithelial degeneration followed by remodeling via the differentiation of regenerative cells in pre-pupal and pupal stages. The changes in the midgut epithelium are paired with the up-regulation of ATG1, ATG6 and ATG8 during the larva-adult transition. Vein, a putative epidermal growth factor involved in regulating epithelial midgut regeneration, is also up-regulated. Autophagy has further been confirmed in sand flies via the presence of autophagosomes residing within the cytoplasmic compartment of the pupal stages. An understanding of the underlying mechanisms of this process should aid the future management of this neglected tropical vector.

Structural and ultrastructural description of larval development in Zungaro jahu.

The Zungaro jahu is an important large catfish of the order Siluriformes that is in danger of extinction due to habitat destruction. Studies on its biology are scarce and the majority relates only to nutrition or parasitology. In order to provide greater information on its morphology and aid husbandry and larviculture studies, the aim of this study was to characterize larval development in Z. jahu from hatching to total yolk absorption. Samples were collected at pre-established times, processed, stained, and analyzed under stereomicroscopy, light microscopy, and scanning electron microscopy. Total yolk absorption was observed by 60 hours post-hatching (hph) at 28.75 ± 0.59°C. The newly hatched larvae showed slightly pigmented body, the outline of the digestive tract, evident eyes, and the first swimming movements. Mouth opening took place at 12 hph and the connection between the oral cavity and the rudimentary intestine was observed at 24 hph. Were analyzed the main larval organs and systems: digestive organs, heart, gill arches, sensory system, thyroid, kidney, and swim bladder. As the larvae grew, these organs became more mature and functional. The development of the sensory and feeding structures was observed at the start of larval development, and thus before depletion of endogenous energy reserves, the strategy for this species is to increase its chances of survival in the environment.

[ULTRASTRUCTURE OF PARENCHYMA IN THE SYNCYTIAL DIGESTIVE SYSTEM IN TURBELLARIA Convoluta convoluta (Acoela].

The paper presents data on the ultrastructure of parenchyma that is involved in the digestion in turbellaria Convoluta convoluta (n = 15). Unusual connections between the nuclear envelope, endoplasmic reticulum and plasma membrane of parenchymal cells were found for the first time, which may indicate the origin of these cell structures. The double trophic role of zooxanthellae in the organism of Convoluta is described.

[D-cells of the gastroenteropancreatic system: development, structure, function and regeneration (history and current state of the problem)].

The present review summarizes the literature data and the results of authors' own research on the development, structure, function and regeneration of D-endocrinocytes of gastroenteropancreatic (GEP) system. The history of the research of these cells is reviewed and its current state of the problem is discussed. The information on the difference of somatostatin-producing D-endocrinocytes from other types of endocrine cells of GAP system is presented, namely, the prevalence of these cells in all the organs of the digestive system (stomach, small and large intestine, pancreas) and other systems of the body, the peculiarities of their structure and regeneration in various species of vertebrate animals and humans in embryonic development, under conditions of normal functioning and in various types of pathology. On the basis of the data on the early differentiation of D-endocrinocytes and their secretion of hormones during embryonic development, structure, cytophysiology and relationships within the general endocrinocyte population, it is suggested that D-endocrinocytes play an important role in the morpho-functional state of GEP system.

Defects in coatomer protein I (COPI) transport cause blood feeding-induced mortality in Yellow Fever mosquitoes.

Blood feeding by vector mosquitoes provides the entry point for disease pathogens and presents an acute metabolic challenge that must be overcome to complete the gonotrophic cycle. Based on recent data showing that coatomer protein I (COPI) vesicle transport is involved in cellular processes beyond Golgi-endoplasmic reticulum retrograde protein trafficking, we disrupted COPI functions in the Yellow Fever mosquito Aedes aegypti to interfere with blood meal digestion. Surprisingly, we found that decreased expression of the γCOPI coatomer protein led to 89% mortality in blood-fed mosquitoes by 72 h postfeeding compared with 0% mortality in control dsRNA-injected blood-fed mosquitoes and 3% mortality in γCOPI dsRNA-injected sugar-fed mosquitoes. Similar results were obtained using dsRNA directed against five other COPI coatomer subunits (α, ß, ß', δ, and ζ). We also examined midgut tissues by EM, quantitated heme in fecal samples, and characterized feeding-induced protein expression in midgut, fat body, and ovary tissues of COPI-deficient mosquitoes. We found that COPI defects disrupt epithelial cell membrane integrity, stimulate premature blood meal excretion, and block induced expression of several midgut protease genes. To study the role of COPI transport in ovarian development, we injected γCOPI dsRNA after blood feeding and found that, although blood digestion was normal, follicles in these mosquitoes were significantly smaller by 48 h postinjection and lacked eggshell proteins. Together, these data show that COPI functions are critical to mosquito blood digestion and egg maturation, a finding that could also apply to other blood-feeding arthropod vectors.

Exsheathment and midgut invasion of nocturnally subperiodic Brugia malayi microfilariae in a refractory vector, Aedes aegypti (Thailand strain).

Exsheathment and midgut invasion of nocturnally subperiodic Brugia malayi microfilariae were analyzed using light and scanning electron microscopy in a refractory vector, Aedes aegypti (Thailand strain). Results showed that exsheathed microfilariae represented only approximately 1% of the total microfilaria midguts dissected at 5-min post-infected blood meal (PIBM). The percentage of exsheathed microfilariae found in midguts progressively increased to about 20, 60, 80, 90, and 100% at 1-, 2-5-, 6-12-, 18-36-, and 48-h PIBM, respectively. Importantly, all the microfilariae penetrating the mosquito midguts were exsheathed. Midgut invasion by the exsheathed microfilariae was observed between 2- and 48-h PIBM. SEM analysis revealed sheathed microfilariae surrounded by small particles and maceration of the microfilarial sheath in the midguts, suggesting that the midguts of the refractory mosquitoes might have protein(s) and/or enzyme(s) and/or factor(s) that induce and/or accelerate exsheathment. The microfilariae penetrated the internal face of the peritrophic matrix (PM) by their anterior part and then the midgut epithelium, before entering the hemocoel suggesting that PM was not a barrier against the microfilariae migrating towards the midgut. Melanized microfilariae were discovered in the hemocoel examined at 96-h PIBM suggesting that the refractory mosquitoes used melanization reactions against this parasite. This study provided evidence that A. aegypti (Thailand strain) has refractory mechanisms against B. malayi in both midgut and hemocoel.

Biochemical, histological and ultrastructural alterations of the alimentary system in the freshwater crab Sinopotamon henanense subchronically exposed to cadmium.

Cadmium (Cd) is one of the toxic metals in the aquatic environment. We investigated the effects of Cd on the digestive enzymes, antioxidant enzymes, malondialdehyde (MDA) content and morphology of the hepatopancreas and intestine in the freshwater crab Sinopotamon henanense. Crabs were exposed to sublethal Cd concentrations of 0, 0.725, 1.450 and 2.900 mg/L for 21 days. After Cd exposure, the activities of maltase, amylase and trypsin of two tissues were lower than the control. The activities of superoxide dismutase, catalase and glutathione peroxidase in the hepatopancreas and intestine were decreased, and the MDA concentration increased in all of the treated groups, over the experimental period. The results of light and transmission electron microscopy showed that 2.900 mg/L of Cd exposure caused profound morphological damages in the hepatopancreas and midgut. After exposure, histological abnormalities of two tissues were discovered, including cellular swelling and necrosis. Additionally, alterations in microvilli, nucleus, mitochondria, rough endoplasmic reticulum as well as Golgi complex in epithelial cells of two tissues were observed. This may be due to antioxidant enzymes activities that were reduced by Cd in the alimentary system of the crabs, and led to membrane lipid peroxidation. The membrane structure was destroyed, and caused further tissue damage, which likely made the alimentary system unable to secrete digestive enzymes, leading to further reduction of digestive enzymes.

Histopathological effects and immunolocalization of periplocoside NW from Periploca sepium Bunge on the midgut epithelium of Mythimna separata Walker larvae.

Periplocoside NW (PSNW) with pregnane glycoside skeleton is a novel insecticidal compound isolated from the root bark of Periploca sepium Bunge. This compound has a potent stomach poisoning activity against several insect pests. In this study, we observed the intoxication symptoms, investigated the histopathological effects and carried out immuno-electron microscopic localization of PSNW on the midgut epithelium of oriental armyworm Mythimna separata Walker larvae for better understanding its action mechanism against insects. Ultrastructural observations showed that cell damages caused by PSNW in the midgut of M. separata larvae are related to the degeneration of brush border microvilli. The dissolution of cytoskeletal structures in the interior and on the surface of microvilli was responsible for the decrease in size and eventual disappearance of microvilli when bubbles of cytoplasmic substances protrude into the midgut lumen of M. separata, thus resulting in cell death. The immuno-electron microscopic localization research showed that gold particle appeared on the microvilli layer of the midgut of M. separate larvae firstly. The density of gold particle gradually added with the time, and finally microvilli layer was destructed severely. Meantime, the gold particles were also presented to the intracellular organelle membrane and the organelles also were destructed. Therefore, we proposed that this membrane system on insect midgut epithelium cells is the initial acting site of PSNW against insects.

MIDGUT ULTRASTRUCTURE OF FOURTH INSTAR OCHLEROTATUS TOGOI (DIPTERA: CULICIDAE).

The ultrastructure of the midgut of fourth instar Ochlerotatus togoi was investigated by light, scanning and transmission electron microscopy. This study was performed to provide information to help devise future control efforts aimed at the larval stages of this vector of filariasis. The fourth instar midgut was approximately 2 mm in length and consisted of three morphologically distinct cell types: epithelial, regenerative, and endocrine cells. There was a monolayer of epithelial cells on the luminal surface of the midgut, with multiple folds of the plasma membrane where it adjoined the basement membrane. Regenerative cells were scattered throughout the basal portion of the epithelium, along with endocrine cells. No evidence of division or differentiation was seen in any of the cell types. Six layers of the peritrophic matrix were observed in the gut lumen which separated ingested food from the midgut epithelial cells. Cytoplasmic protrusions were seen in many areas of the luminal midgut surface and numerous autophagosomes were seen in the epithelial cells of both early and late fourth instar larvae, suggesting autophagy is involved in the degeneration process of the midgut in preparation for pupation. This study provides a basis for understanding normal Oc. togoi larval midgut development. Further studies are needed to determine the factors that control larval growth and the nutritional state. Such information could be used to reduce adult fecundity and develop biological control mechanisms.

Gut expansion and contraction in the predatory soil mite Pergamasus longicornis (Mesostigmata: Parasitidae): a stiff system.

Mite digestive processes are inferred from gut expansion and contraction time in the free-living predatory soil mite Pergamasus longicornis (Berlese), estimated using a temporal series of histological sections. Gut regions (bar the rectal vesicle) behave broadly in unison for rapid initial filling (ingestion half-life about 2-3 min; max 8 min), but behave heterogeneously when slowly emptying (digestion/egestion half-life from about 2-3 h; max 8.5 h). Anterior gut regions fill and empty the earliest. Posterior gut regions take the longest to fill and to empty. Switching first from filling-predominating to emptying-predominating in the gut occurs around 2 h from the start of feeding. Median time for the initial completion of gut filling and for the commencement of gut emptying is 10 min and 12.5 h, respectively, from the start of feeding. Three phases of gut changes are critically discussed: rapid filling, concentration by fluid loss (via coxal glands), and slow emptying. Independent corroboration of coxal droplet formation is included. Predictions to confirm or refute postulated mechanisms of salivary, coxal or rectal water balance are given. Overall total gut filling (ingestion) plus gut emptying (digestion/egestion) time in this poikilotherm is approximately 29-52.5 h (1+ - 2+ days) at room temperature from the start of feeding on large dipteran prey ([Formula: see text] gut emptyings per day). Pergamasus longicornis exhibits the stiff digestive system of an intermittent 'bolus' feeder.