Ultrastructural studies of the formation of the egg capsule in the hermaphroditic species, Isohypsibius granulifer granulifer Thulin, 1928 (Eutardigrada: Hypsibiidae)

The egg capsule of Isohypsibius granulifer granulifer Thulin 1928 (Eutardigrada: Hypsibiidae) is composed of two shells: the thin vitelline envelope and the multilayered chorion. The process of the formation of the egg shell begins in middle vitellogenesis. The I. g. granulifer vitelline envelope is of the primary type (secreted by the oocyte), but the chorion should be regarded as a mixed type: primary (secreted by the oocyte), and secondary (produced by the cells of gonad wall). During early choriogenesis, the parts of the chorion are produced and then connected into a permanent layer. The completely developed chorion consists of three layers: (1) the inner, medium electron dense layer; (2) the middle labyrinthine layer; (3) the outer, medium electron dense layer. After the formation of the chorion, a vitelline envelope is secreted by the oocyte.     

Autophagy as the cell survival in response to a microsporidian infection of the midgut epithelium of Isohypsibius granulifer granulifer (Eutardigrada: Hypsibiidae)

The midgut epithelial cells of many invertebrates may possess microorganisms which act as symbionts or pathogens (bacteria, microsporidia, viruses). During our previous studies on Isohypsibius granulifer granulifer Thulin, 1928 (Tardigrada, Eutardigrada), which examined alterations of the midgut epithelium during oogenesis, we found that some of the specimens were infected with microsporidia. All stages of pathogens occurred in the cytoplasm of the digestive cells in the midgut epithelium of I. g. granulifer that were infected with microsporidia: meronts, sporonts, sporoblasts, and spores. The cytoplasm of the digestive cells was rich in mitochondria, cisterns of rough endoplasmic reticulum (RER), and Golgi complexes. Autophagy in the digestive cells of the dorsal midgut was much more intensive in comparison with noninfected specimens. Membranes of phagophores surrounded the pathogens forming autophagosomes. These latter structures fused with lysosomes forming autolysosomes and residual bodies appeared. Neither glycogen granules nor droplets of varying electron density, which accumulated in digestive cells during vitellogenesis and choriogenesis, appeared in individuals with microsporidia. While the midgut epithelium in noninfected specimens takes part in vitellogenesis and choriogenesis, in infected specimens, midgut cells are involved in the process of autophagy as a survival strategy.     

Yolk formation in Isohypsibius (Eutardigrada)

Summary In Isohypsibius granulifer, yolk is autosynthesized. The Golgi apparatus is mainly responsible for the formation of yolk, which consists of irregular platelets with heterogeneous contents and a diameter of about 1 m. Dense globules, 300 nm in diameter, are visible among yolk platelets. These develop in the vesicles of the rough endoplasmic reticulum. The genesis of these vesicles is associated with the outer membrane of the nuclear envelope, which forms blebs intensively during previtellogenesis and early vitellogenesis. The developing oocytes are assisted by nurse cells, to which they are jointed by cytoplasmic bridges. For every oocyte, there are a number nurse cells, which are sister cells of the oocyte. In addition to rRNA, nurse cells transfer to the oocyte lipids, platelets of yolk formed in their cytoplasm, mitochondria and cortical granules.     

Ultrastructural changes in the midgut epithelium in Podura aquatica L. (Insecta, Collembola, Arthropleona) during regeneration

Data considering the degeneration and regeneration of the midgut epithelium in the primitive wingless insects, such as Collembola, are rather poor. Also information, which treats the regenerative cells as the primordial cells, is poorly known. The midgut epithelium of Podura aquatica L. (Insecta, Collembola, Arthropleona) is formed by the epithelial and regenerative cells. The epithelial cells show distinct regionalisation in the organelles distribution. The ultrastructure of the basal, perinuclear and apical regions of the epithelial cells is described. As in insects without Malpighian tubules, structures which resemble urospherites occur in the cytoplasm of the epithelial cells. After degeneration of the entire midgut epithelium, a new epithelium is formed from regenerative cells. During the process of regeneration, the degenerated epithelium gradually is separated from the basal lamina by the newly formed one. Finally, the detached epithelium is moved into the midgut lumen. Regenerative cells play a role of primordial cells during epithelial regeneration.     

Tardigrada of the Revdalen (Spitsbergen) with the descriptions of two new species: Bryodelphax parvuspolaris (Heterotardigrada) and Isohypsibius coulsoni (Eutardigrada)

Despite a century long history of research, tardigrade fauna of the Svalbard Archipelago remains poorly known. In order to deepen our knowledge of tardigrade biodiversity in the Arctic, we collected forty-one moss and lichen samples from the Revdalen and on the south-east slopes of the Rotjesfjellet (Spitsbergen, Svalbard Archipelago) in June 2010. In these samples, twenty-five tardigrade species were found, including two new for science: Bryodelphax parvuspolaris sp. nov. and Isohypsibius coulsoni sp. nov. B. parvuspolaris sp. nov. belongs to the weglarskae group but differs from all other species of the group by a unique configuration of ventral plates. I. coulsoni sp. nov. differs from the most similar species I. ceciliae Pilato and Binda, 1987 mainly by the absence of ventral sculpture. Two additional species, Milnesium asiaticum Tumanov, 2006 and Diphascon (Adropion) prorsirostre Thulin, 1928, are recorded from the Svalbard Archipelago for the first time.     

Phylogenetic Relationships in the Macrobiotidae (Tardigrada: Eutardigrada: Parachela)

Characters of the claws, buccal-pharyngeal apparatus and ultrastructure of the cuticle have been examined to identify phyletic relationships within the Macrobiotidae. Several apomorphies have been identified. The resulting cladogram shows two main phyletic lines within the family, corresponding to the subfamilies Macrobiotinae and Murrayinae. The relationships among the genera of the Murrayinae are resolved, whereas in the Macrobiotinae five main phyletic lines are identified, whose relationships are still unknown. Pseudodiphascon and Macrobiotus appear to be polyphyletic genera.     

Photokinesis of Macrobiotus hufelandi (Tardigrada, Eutardigrada)

Macrobiotus hufelandi Schultze, 1834 was tested for response to light. Size groups of less than 120 m and of 120 m or more were used. It was found that the smaller, younger size group exhibited a statistically significant negative response to light. This is hypothesized to function in conservation of body moisture and be more important to the smaller individuals because of the greater surface area-to-volume ratio. An experiment on response to gravity was done in an effort to determine if the negative photokinesis was the total result of a more complex type of behavior but this experiment did not detect any significant response to gravity in either of the size groups. Observation of the bacterial trails left by the tardigrades indicates that this behavior is not negative phototaxis, but rather negative photokinesis which is a non-directed, random movement in which the animal either increases its speed or changes its direction when exposed to light.     

Cuticle structure and systematics of the Macrobiotidae (Tardigrada, Eutardigrada)

The cuticle of tardigrades is characterized by three main layers: epicuticle, intracuticle and procuticle. Pillars are present in the epicuticle of almost all heterotardigrades, but these structures are also known in a few species of eutardigrades. The apparent heterogeneity of the cuticular ultrastructure in several species of the Macrobiotidae (Eutardigrada) prompted us to analyse the structure of the cuticle in this family. Eleven species in several genera were investigated with light and/or electron microscopy. All the species of the genera Murrayon and Dactylobiotus showed pillars in the epicuticle, whereas the examined species of Macrobiotus , Richtersius and Xerobiotus completely lacked pillars. Therefore, in the Macrobiotidae, in contrast to what appears with light microscopy, the cuticle is homogeneous within each genus examined at the electron microscopic level. Considering the absence of pillars in the Macrobiotidae a synapomorphy, we propose the erection of two new subfamilies. Macrobiotinae subfam. n. is characterized by the absence of pillars in the epicuticular layer and includes, in addition to the genera Macrobiotus, Xerobiotus and Richtersius, the genera Pseudohexapodibius, Adorybiotus, and probably also Minibiotus, Calcarobiotus and Pseudodiphascon . Murrayinae subfam. n. is characterized by the presence of pillars in the epicuticular layer and includes the genera Murrayon, Dactylobiotus and, probably, Macroversum.     

Ultrastructural changes in the midgut epithelium of the first larva of Allacma fusca (Insecta, Collembola, Symphypleona)

Abstract. In the newly hatched larva in Allacma fusca , the midgut epithelium was fully developed and formed by flattened epithelial cells surrounding the yolk mass in the midgut lumen. Immediately after hatching, the first larva began to feed; the migut lumen was filled with the yolk mass and food (mainly algae). Regenerative cells typical of the developing midgut epithelium of many insects were not observed. Initially, midgut cells of the larva were cuboidal but became columnar in shape with distinct regionalization in the distribution of cell organelles. Furthermore, urospherites appeared in the midgut cell cytoplasm, i.e., structures characteristic for the midgut epithelium of insects having no Malpighian tubules. As a result, cells with the capacity for digestion, absorption, and excretion were observed to be completely formed in the first larval stage.     

Form and function of the feeding apparatus in Eutardigrada (Tardigrada)

Tardigrade feeding apparatus is a complex structure with considerable taxonomic significance that can be schematically divided into four parts: buccal ring, buccal tube, stylet system, and pharynx. We analyzed the fine morphology and the tridimensional organization of the tardigrade buccal?Cpharyngeal apparatus in order to clarify the relationships between form and function and to identify new characters for systematic and phylogenetic studies. We conducted a comparative analysis of the cuticular structures of the buccal?Cpharyngeal apparatuses of twelve eutardigrade species, integrating data obtained by SEM and LM observations. Morphological diversity was observed and new cuticular structures such as the stylet coat of the stylet system were identified. The synthesis of the buccal?Cpharyngeal apparatus during molting was also analyzed obtaining a clear developmental sequence of its resynthesis. These findings lead us to redefine the previous interpretations of the functioning mechanisms of the buccal?Cpharyngeal apparatus and provide a more specific relationship between tardigrade diet and the anatomy of their feeding apparatuses. In addition, the detection by energy-dispersive X-ray spectroscopy of calcium in the stylets, buccal tube, and placoids of eutardigrade species (i.e., Milnesium tardigradum, Paramacrobiotus richtersi) indicates that CaCO₃ incrustations are not an exclusive feature of heterotardigrades and lead to suppose that this trait was present in the ancestors of both classes.     

Paramacrobiotus lachowskae,a new species of Tardigrada from Colombia (Eutardigrada: Parachela: Macrobiotidae)

In this article we describe a new Paramacrobiotus species of the areolatus group by means of integrative taxonomy. Together with the detailed morphological and morphometric data (obtained from light and scanning electron microscopy) we also provide DNA sequences of four universal molecular markers used in tardigrade taxonomy (three nDNA fragments: 18S rRNA, 28S rRNA, ITS-2, and one mtDNA fragment: COI). The animals of Paramacrobiotus lachowskae sp. nov. are similar to several species of the areolatus complex and the eggshell ornamentation is similar to that of two species of the richtersi group. Therefore, the new species can be easily distinguished from the areolatus group species by egg morphology (dome-like, wrinkled processes with long flexible spines/filaments covered by fine short hairs) and from species of the richtersi complex by the absence of the microplacoid. Paramacrobiotus lachowskae sp. nov. is the 43rd species reported from Colombia and fifth Paramacrobiotus species from this South American country.

http://zoobank.org/urn:lsid:zoobank.org:pub:FEDC11D3-DCCF-4699-A704-CE6DEA1CDED4     

Ultrastructural studies of midgut epithelium formation in Lepisma saccharina L. (Insecta, Zygentoma)

At the end of embryogenesis of Lepisma saccharina L. (Insecta, Zygentoma), when the stomodaeum and proctodaeum are completely formed, the midgut epithelium is replaced by the primary midgut, a yolk mass is surrounded by a cell membrane. Midgut epithelium formation begins in the 1st larval stage. Energids migrate toward the yolk periphery and aggregate just beneath the cell membrane. They are gradually enclosed by cell membrane folds of the primary midgut. Single cells are formed. Succeeding energids join just formed cells. Thus, groups of cells, regenerative cell groups, are formed. Their number gradually increases. The external cells of the regenerative cell groups transform into epithelial cells and their basal regions spread toward the next regenerative cell groups. Epithelial cells of neighboring regenerative cell groups join each other to form the epithelium. At the end of the 2nd larval stage, just before molting, degeneration of newly the formed epithelium begins. Remains of organelles and basal membrane occur between the regenerative cell groups. The new epithelium is formed from the regenerative cell groups, which are now termed stem cells of the midgut epithelium.     

Ultrastructural studies on the midgut epithelium and digestion in the female tickArgas (Persicargas) arboreus (Ixodoidea: Argasidae)

The ultrastructure of the midgut epithelium and digestion in the female tickArgas (Persicargas) arboreus are described before and after feeding, up to oviposition. The epithelium consists of secretory cells, digestive cells (DI and DII), and regenerative cells which may differentiate into any of the other cell types. In unfed ticks, the midgut wall consists mainly of type DII digestive cells retained from a previous feeding, and a few regenerative cells. Within 3 days after the tick feeding, haemolysis of the host blood components occurs in the midgut lumen. Secretory cells, the first differentiation of the regenerative cells, are presumed to produce a haemolysin and an anticoagulant which are released by merocrine and holocrine secretions. The DII cells seen in unfed ticks, and secretory cells which have completed their secretory cycle, start to have a specialized surface for endocytosis characteristic of type DI digestive cells. From 5 to 7 days after feeding up to the female oviposition, type DI cells which have completed their endocytosis are transformed into type DII digestive cells specialized for intracellular digestion and the storage of reserve nutrients required by the tick for long starvation. The various phases of the digestive cycle are considered according to ultrastructural changes of the midgut epithelium.     

A new species of Tardigrada (Eutardigrada: Milnesiidae): Milnesium krzysztofi from Costa Rica (Central America)

Abstract

A new eutardigrade species Milnesium krzysztofi sp. nov. is described from Costa Rica. M. krzysztofi sp. nov. differs from the most similar Milnesium katarzynae Kaczmarek et al., 2004 mainly by the presence of spurs on internal claws I–III and on external claw IV.     

The ovary structure, previtellogenic and vitellogenic stages in parthenogenetic species Dactylobiotus dispar (Murray, 1907) (Tardigrada: Eutardigrada)

The reproductive system of Dactylobiotus dispar consists of the ovary and the oviduct that opens into the rectum. The sack-like ovary is filled with the developing oocytes, which are assisted by the trophocytes. In D. dispar, the mixed vitellogenesis takes place. One part of the yolk material is produced inside the oocyte (autosynthesis), the second part is absorbed by micropinocytosis while the third part is synthesized in the trophocytes and is transported to the oocytes through the cytoplasmatic bridges. Moreover, rRNA, lipids and mitochondria are transfered from the trophocytes to the oocytes. The histochemical researches show that the reserve material accumulated in the oocytes contains proteins, polysaccharides and lipids.     

Ultrastructural features of the midgut epithelium of females Lutzomyia intermedia (Lutz & Neiva, 1912) (Diptera: Psychodidae: Phlebotominae).

A morphological study of the midgut of Lutzomyia intermedia, the primary vector of cutaneous leishmaniasis, in southeast Brazil, was conducted by light, scanning and transmission electron microscopy. The midgut is formed by a layer of epithelium of columnar cells on a non-cellular basal lamina, under which there is a musculature, which consists of circular and longitudinal muscular fibers. A tracheolar network is observed surrounding and penetrating in the musculature. Females were examined 12, 24, 48, 72 h and 5 days following a blood meal and were analyzed comparatively by transmission electron microscopy with starved females. In starved females, the epithelium of both the anterior and posterior sections of the midgut present whorl shaped rough endoplasmic reticulum. The posterior section does not present well-developed cellular structures such as mitochondria. Observations performed at 12, 24, 48 and 72 h after the blood meal showed morphological changes in the cellular structures in this section, and the presence of the peritrophic matrix up to 48 h after the blood meal. Digestion is almost complete and a few residues are detected in the lumen 72 h after blood feeding. Finally, on the 5th day after the blood meal all cellular structures present the original feature resembling that seen in starved sand flies. Morphometric data confirmed the morphological observations. Mitochondria, nuclei and microvilli of midgut epithelial cells are different in starved and blood fed females. The mitochondria present a similar profile in the epithelium of both the anterior and posterior section of the midgut, with higher dimension in starved females. The cell microvilli in the posterior section of the midgut of starved females are twice the size of those that had taken a blood meal. We concluded that there are changes in the midgut cellular structures of L. intermedia during the digestion of blood, which are in agreement with those described for other hematophagous diptera.