Gastrodermis ultrastructure of the different life stages of the polyopisthocotylean monogenean gill parasite Discocotyle sagittata

The polyopisthocotylean Discocotyle sagittata is a blood-feeding monogenean that infects the gill lamellae of rainbow trout, Oncorhynchus mykiss, and brown trout, Salmo trutta. The ultrastructure of their alimentary tract, at different stages of the life cycle, was previously unknown. Here, we show that the gastrodermis of the oncomiracidium, subadult, and adult D. sagittata follows the same structural organization as that of other blood-feeding polyopisthocotyleans, being composed of digestive cells alternating with a connecting syncytium. Digestive cells of the oncomiracidium are found in three developmental forms: undifferentiated, developing differentiated, and differentiated (presumably functioning) cells whereas those of adult and subadult are present in a single functioning state with variable size and content. The apical cytoplasm of adult digestive cells forms conical outgrowths, a feature which is absent in the oncomiracidium. The connecting syncytium of the oncomiracidium has no evidence of metabolic activity, while that of adult and subadult is metabolically active. The lamellae of the connecting syncytium of adults and subadults are more numerous and larger, and their terminal portions are expanded, compared with those of the oncomiracidium. Parallel, tubular, membranous structures are characteristic of the apical cytoplasm of the connecting syncytium of the oncomiracidium. Luminal lamella in the oncomiracidium, subadult, and adult form balloon-like structures enclosing some luminal contents, but those of the oncomiracidium are larger, bounded by nucleated cytoplasmic layer, and enclose more luminal contents. The possible functions of these structures and mechanism of digestion in both oncomiracidium and adult are discussed.

Mialostatin, a Novel Midgut Cystatin from Ixodes ricinus Ticks: Crystal Structure and Regulation of Host Blood Digestion

The hard tick Ixodes ricinus is a vector of Lyme disease and tick-borne encephalitis. Host blood protein digestion, essential for tick development and reproduction, occurs in tick midgut digestive cells driven by cathepsin proteases. Little is known about the regulation of the digestive proteolytic machinery of I. ricinus. Here we characterize a novel cystatin-type protease inhibitor, mialostatin, from the I. ricinus midgut. Blood feeding rapidly induced mialostatin expression in the gut, which continued after tick detachment. Recombinant mialostatin inhibited a number of I. ricinus digestive cysteine cathepsins, with the greatest potency observed against cathepsin L isoforms, with which it co-localized in midgut digestive cells. The crystal structure of mialostatin was determined at 1.55 Å to explain its unique inhibitory specificity. Finally, mialostatin effectively blocked in vitro proteolysis of blood proteins by midgut cysteine cathepsins. Mialostatin is likely to be involved in the regulation of gut-associated proteolytic pathways, making midgut cystatins promising targets for tick control strategies.

A physiologic overview of the organ-specific transcriptome of the cattle tick Rhipicephalus microplus

To further obtain insights into the Rhipicephalus microplus transcriptome, we used RNA-seq to carry out a study of expression in (i) embryos; (ii) ovaries from partially and fully engorged females; (iii) salivary glands from partially engorged females; (iv) fat body from partially and fully engorged females; and (v) digestive cells from partially, and (vi) fully engorged females. We obtained > 500 million Illumina reads which were assembled de novo, producing > 190,000 contigs, identifying 18,857 coding sequences (CDS). Reads from each library were mapped back into the assembled transcriptome giving a view of gene expression in different tissues. Transcriptomic expression and pathway analysis showed that several genes related in blood digestion and host-parasite interaction were overexpressed in digestive cells compared with other tissues. Furthermore, essential genes for the cell development and embryogenesis were overexpressed in ovaries. Taken altogether, these data offer novel insights into the physiology of production and role of saliva, blood digestion, energy metabolism, and development with submission of 10,932 novel tissue/cell specific CDS to the NCBI database for this important tick species.

Ultrastructure of the Bacteriocytes in the Midgut of the Carpenter ant Camponotus rufipes: Endosymbiont Control by Autophagy

The carpenter ant Camponotus rufipes has intracellular bacteria in bacteriocytes scattered in the midgut epithelium, which have different amounts of endosymbionts, according to the developmental stages. However, there are no detailed data about the midgut cells in adult workers. The present work aimed to evaluate the morphology and cellular events that coordinate the abundance of endosymbionts in the midgut cells in C. rufipes workers. The midgut epithelium has digestive cells, bacteriocytes, and cells with intermediate morphology. The latter is similar to bacteriocytes, due to the abundance of endosymbionts, and similar to digestive cells, due to their microvilli. The digestive and intermediate cells are rich in autophagosomes and autolysosomes, both with bacteria debris in the lumen. These findings suggest that midgut cells of C. rufipes control the endosymbiont level by the autophagy pathway.

Morphological characteristics of respiratory and digestive organs of Roman snail (Helix pomatia L., 1758)

The aim of this work was to study the structure of lung and hepatopancreas of Roman snail (Helix of pomatia of L., 1758). The study found that the lung occupies the lower turn of shell and presented by a saccate cavity, in the wall of that there are a kidney and heart with a pericardium, and also a rectum and ureter pass. An external surface of lungs covered by a shell and covered by an epidermis. An internal surface is covered by a flat ciliated epithelium and forms numerous folds in which pulmonary vessels and lacunae are accommodated. The branches of pulmonary vein have a thick muscular wall, that consists of circular and longitudinal muscular layers. An internal surface of lungs covered by the layer of mucus. Inhalation and exhalation are carried out due to reduction and relaxation of muscles of dorsal wall of the body that is named a “diaphragm”. Gas exchange occurs through the hemolymphatic capillaries of the lung wall. Respiratory motions take place not rhythmically, but through the different intervals of time depending on a requirement in oxygen. The frequency of pneumostome closing and opening is typically one time in a minute. At subzero humidity of atmospheric air of pneumostome closed by a mantle, and also one (or a few) epiphragms. The hepatopancreas (“liver” or liver gland) is in the upper rotation of the sink and formed by two parts: right and left, from which two liver ducts enter into the stomach respectively. The liver gland consists of many acinuss, surrounded by connecting tissue, that contains small number of muscular fibres. Calcium cells have a pyramidal form and usually do not reach the lumen of the acinus. Cytoplasm of calcium cells contains inclusions: grains of phosphoricacid lime and drops of fat. The digestive cells of the hepatopencreas are more elongated, often clavicular. Сytoplasm of digestive cells is loose and vacuolated and contain inclusions of yellow-green color. Enzyme cells on histopreparations are difficult to distinguish from digestive ones. They contain transparent vacuoles with a large round inclusion of yellow-green color, which consists of a cluster of several grains of different sizes. Hepatopancreas performs the following functions: secretory (enzyme cells), absorption and intracellular digestion (digestive cells), preservation of nutrients and calcium (calcium cells), and also excretory function.

Ultrastructure of the midgut epithelium in three species of Macrobiotidae (Tardigrada: Eutardigrada: Parachela)

Three species of Macrobiotidae, Macrobiotus polonicus, Macrobiotus diversus and Macrobiotus pallarii, were selected for analysis of the fine structure of the midgut epithelium. They are gonochoric and carnivorous species that live in wet terrestrial and freshwater environments. The ultrastructure of the midgut epithelium of the investigated Macrobiotidae species was analysed in both males and females. Their digestive system is composed of fore- and hindguts that are covered by a cuticle, and the middle region, termed the midgut. It is lined with a simple epithelium that is formed by digestive cells that have a distinct brush border. Crescent-shaped cells that form an anterior ring in the border between the fore- and midgut were detected. The ultrastructure of the intestinal epithelium of the examined species differs slightly depending on sex. The digestive cells of the posterior segment of the intestine contain numerous lipid droplets, which are the reserve material. We concluded that the digestive cells of the Macrobiotidae midgut are responsible for its intracellular digestion owing to endocytosis. They also participate in the extracellular digestion owing to merocrine secretion (exocytosis). However, the midgut is not the main organ that accumulates reserve material. Additionally, the midgut epithelium does not participate in oogenesis.

Autophagy and Apoptosis in the Midgut Epithelium of Millipedes

The process of autophagy has been detected in the midgut epithelium of four millipede species: Julus scandinavius, Polyxenus lagurus, Archispirostreptus gigas, and Telodeinopus aoutii. It has been examined using transmission electron microscopy (TEM), which enabled differentiation of cells in the midgut epithelium, and some histochemical methods (light microscope and fluorescence microscope). While autophagy appeared in the cytoplasm of digestive, secretory, and regenerative cells in J. scandinavius and A. gigas, in the two other species, T. aoutii and P. lagurus, it was only detected in the digestive cells. Both types of macroautophagy, the selective and nonselective processes, are described using TEM. Phagophore formation appeared as the first step of autophagy. After its blind ends fusion, the autophagosomes were formed. The autophagosomes fused with lysosomes and were transformed into autolysosomes. As the final step of autophagy, the residual bodies were detected. Autophagic structures can be removed from the midgut epithelium via, e.g., atypical exocytosis. Additionally, in P. lagurus and J. scandinavius, it was observed as the neutralization of pathogens such as Rickettsia-like microorganisms. Autophagy and apoptosis ca be analyzed using TEM, while specific histochemical methods may confirm it.