The structure and ultrastructure of the egg capsules of stoneflies of the genus Isoperla (Insecta,Plecoptera, Perlodidae)

The egg capsules of five systellognathan stoneflies species representing the genus Isoperla (Plecoptera, Perlodidae) have been investigated using light and electron microscopes (SEM and TEM). We consider the structural modifications of egg coverings (egg capsules, eggshells) like: a shape of a capsule, presence of structures fixing an egg to the substratum under water, intrachorionic aeropylar system facilitating gas exchange as a factor adapting an egg/embryo to the environment. The structures protecting eggs against desiccation during a female flight before oviposition into water as well as against the dangers of external mechanical injury caused by turbulences in rapid water currents of mountain streams are described and discussed. The ground plan of the egg capsule in arctoperlarian stoneflies is also discussed.     

Comparative ultrastructure of the fertilized egg envelope in Corydoras adolfoi and Corydoras sterbai,Callichthyidae, Teleostei

In teleost, the structural characteristics of fertilized egg and egg envelope are very important for classification of genus or species. The structures of fertilized egg and egg envelope from Corydoras adolfoi and Corydoras sterbai, Callichthyidae, Siluriformes in teleost were examined by scanning and transmission electron microscopes to confirm whether these morphological structures have specificities of species and family or not. The fertilized eggs of C. adolfoi and C. sterbai were non‐transparent, spherical, demersal, and strong adhesive. There were no structural differences between two species through the light microscope. The size of the fertilized eggs of C. adolfoi was 1.95 ± 0.03 mm (n = 20), and that of C. sterbai was 1.92 ± 0.03 mm (n = 20). The perivitelline space was almost not developed in both species. In both species, the adhesive protuberances structures were on the outer surface of egg envelope. And fibrous structures were specially located at attachment part of spawning bed. And the egg envelope consisted of two layers, an inner lamellae layer and an outer strong adhesive layer with high electron dense protuberances structures in cross section. Consequentially, the fertilized eggs, outer surface on the egg envelope and cross section of egg envelope have identical structure. So, these structural characteristics of fertilized eggs and egg envelope show genus Corydoras specificity.     

Reproductive performance of the Mesa silverside (<i>Chirostoma jordani</i> Woolman, 1894) under natural and controlled photoperiods

Chirostoma jordani is a native annual species inhabiting lacustrine waters of the Central Mexico Plateau. It is widely distributed and is currently facing high environmental pressures. Five experiments were performed to study the reproductive performance of this species. Four of the experiments were conducted in 270-L indoor recirculation tanks. Two males and one female at the first stage of reproduction were included in each test. A photoperiod of 14 light hours and 10 dark hours was used. In a fifth experiment, 10 females and 15 males were kept in an outdoor 3,000-L recirculation tank under natural photoperiod. The number of spawns, fertilised eggs and 30-day-old juveniles were counted and the survival rate was calculated. The results indicated significant differences (P<0.05) between treatments. Higher spawn numbers and greater egg production were observed under controlled photoperiod, and higher numbers of juveniles and a higher survival rate were observed under natural photoperiod. The trials exhibited different patterns of egg production during the experiment. The egg production in the natural-photoperiod trials followed a polynomial curve model. In contrast, the trials under the controlled photoperiod showed an irregular pattern of increases and decreases in egg production.     

Egg envelopes of Armadillidium vulgare (Latreille, 1804) (Crustacea,Isopoda Oniscidea): Ultrastructure and lectins binding

The ultrastructural study carried out on (a) oocytes of Armadillidium vulgare during vitellogenesis, (b) mature eggs taken from the ovaries during the parturial moult of the posterior half of the body, and (c) fertilized eggs collected within a few hours of their release into the brood pouch, has clearly demonstrated that before the fertilization the chorion is the only envelope present in the egg of oniscidean isopods. In the mature eggs, the chorion appears as a uniformly electron‐dense lamina, about 0.4–0.5 µm thick, which does not show any specialized area. A second envelope, described by other authors as vitelline envelope, is formed above the oolemma only right after fertilization and appears separated from the chorion by a space full of liquid. The ways in which the genesis of this envelope is realized are not yet clear; it could be interpreted rather as a fertilization membrane. The investigations carried out with the aid of a battery of FITC‐lectins have highlighted the presence at the chorion surface of unfertilized eggs of various saccharide residues distributed in uniform way. No significant change was observed in the pattern of lectins binding to the chorion of eggs taken from the brood pouch, thus demonstrating how, after the fertilization, no significant rearrangement in the distribution of saccharide residues present on the egg surface occurs in A. vulgare. The ways in which, therefore, the recognition, the binding and the entry of the peculiar sperm of oniscidean isopods into the egg occur, still remain all to be deciphered. Microsc. Res. Tech. 79:792–798, 2016. © 2016 Wiley Periodicals, Inc.     

Serial sectioning,electron microscopy,and three-dimensional reconstruction of fine nerve fibres and other extended objects

Two different techniques for three-dimensional reconstructions of nerve fibres or other extended fine tissue structures were developed to study the afferent innervation of the cat's knee-joint capsule. The re-embedding technique starts with series of semithin sections for light microscopic reconstructions. In a second step, selected semithin sections can be re-embedded for ultramicrotomy to examine ultrastructural details. This method offers the possibility to investigate fine structures over a distance of several hundred micrometres without any loss of ultrastructural information. The serial section-ESI technique is based on the electron spectroscopic imaging of semithin sections. Small tissue blocks are cut into series of either semithin or alternate semi- and ultrathin sections which can be directly used for a complete ultrastructural investigation. Finally, true-to-scale three-dimensional reconstructions are performed by graphical techniques or computer-aided methods.     

Research of bionic design on tools with rostrum of cyrtotrachelus bugueti guer (coleoptera: curculionidae)

To improve the torsional resistance capability of multilayered composite pipes, a bionic design was presented based on the rostrum of Cyrtotrachelus Bugueti Guer. Morphology and mechanical properties of the weevil's rostrum were studied. Scanning electron microscope (SEM) was employed to observe the macro‐/microstructure and inner structure. Nanoindentation was carried out to measure the elastic modulus and the hardness of the biological materials. Based on the structure of rostrum, a mathematical model of multilayered biomimetic pipe was established with the statistical methods. Then, the 3D biomimetic cylinder was built and the torsional resistance capability was analyzed using the finite element analysis. From the SEM and nanoindentation, it was found that the rostrum is a cylindrical hollow multilayered structure composed by three materials. In the transverse section of rostrum, the materials' moduli are 3.07, 3.15, and 8.64 GPa, the hardness are 172.71, 125.32, and 278.99 MPa, respectively. In the longitudinal section, the moduli are 3.01, 4.35, and 7.66 GPa, the hardness are157.46, 149.15, and 253.51 MPa, respectively. The results of simulation showed that the distributional type of lamination has a significant effect on the torsional resistance capability of multilayered structure. The biomimetic structure, which imitates the lamination of rostrum, is superior in the aspect of torsional resistance compared with the nonlaminated or uniform laminated structure. Additionally, the bionic method is feasible and efficient.     

Morphology and distribution of antennal sensilla in egg parasitoid,Ooencyrtus nezarae (Hymenoptera: Encyrtidae)

Morphology of antennal sensilla and their distribution were investigated in adults of Ooencyrtus nezarae, an egg parasitoid of Riptortus pedestris, using scanning electron microscopy. Male antennae was found to be significantly greater in overall length than female antennae. The antenna of O. nezarae was composed of the radicula, scape, pedicel, funicle and clava in both sexes, with seven types of sensilla identified: sensillum trichodea; s. finger‐like; s. placoidea; s. chaetica; s. basiconica; s. coeloconica, and s. campaniform. They occur in varying number and distribution along the antennae. Two sensillum types were further categorized into additional subtypes, with two subtypes in s. trichodea and three in s. chaetica. Among all characterized sensilla, s. trichodea subtype 1 and s. placoidea were multiporous, indicating that the primary function of these sensilla is olfactory. Sensillum trichodea was the most abundant sensillum type on the antennae of both sexes. Sexual dimorphism was only observed from the subtype 1 sensilla of s. trichodea in males and the subtype 3 sensilla of s. chaetica in females. The morphological information established in our study may provide useful information for further investigations in sensory physiological function of each morphological type of sensilla and their related behavior in this egg parasitoid.     

General morphology and ultrastructure of the female reproductive apparatus of Trichomalopsis shirakii crawford (Hymenoptera,Pteromalidae)

The morphology and ultrastructure of the female reproductive system were examined for a larval–pupal parasitoid Trichomalopsis shirakii Crawford of Oulema oryzae Kuwayama using light and electron microscopes. The reproductive system includes two ovaries, two pairs of accessory glands, an unbranched venom gland, a large venom reservoir and a Dufour gland. Each ovariole contains follicles and oocytes at different stages of maturation. A fibrous layer covers the surface of mature egg. The accessory glands are made up of a layer of secretory cells surrounded by muscle fibers. In these secretory cells, numerous mitochondria, electron‐dense secretory granules and vesicles filled with dense granular particles are present. These granular particles appear as virus‐like particles (VLPs). The venom gland consists of a single layer of secretory cells which are organelle rich with abundant rough endoplasmic reticulum, mitochondria and vesicular organelles, a layer of duct cells and an inner intima. The reservoir consists of a muscular sheath, epidermal cells with few organelles and an intima layer. The Dufour gland has a relatively large lumen surrounded by a single layer of columnar epithelial cells which are characterized by clusters of smooth endoplasmic reticulum and lipid droplets. Aside from the venom, the fibrous layer coating the egg and the granular particles which may be VLPs have been discovered in our study. They may serve as one of the parasitoid‐associated factors in their host–parasitoid relationship and play a role in host immune suppression. Microsc. Res. Tech. 79:625–636, 2016. © 2016 Wiley Periodicals, Inc.     

The structure and formation of the egg membranes in Nasonia vitripennis (Walker) (Hymenoptera, Pteromalidae)

A histochemical and fine structural study of the vitelline membrane and chorion was carried out. Following the uptake of protein at the oöcyte plasma membrane during yolk synthesis, the egg membranes are laid down in sequence in the intercellular space between the oöcyte and the follicle cells. The follicular epithelial cells alone contribute towards their formation. The structure of the chorion is much simpler than that of Bombus, the only other hymenopteran previously investigated.     

Fracture load and microcrack comparison of crowns manufactured from tooth‐shaped and traditional blocks

This study intended to analyze microcracks and fractographic markings on the surface of all ceramic crowns after milling and compare the fracture loads. 90 crowns were manufactured from two feldspathic (Priticrown‐Pr and Vita Mark II‐Vi) and a lithium disilicate (EmaxCAD‐Em) blocks (n = 30). Two groups (n = 15) were prepared for each ceramic. In the first group, crowns were analyzed twice via the fluorescent penetrant method for microcrack detection, after the manufacturing process and thermal cycles. The load to fracture test was applied at a crosshead speed of 0.5 mm/min until catastrophic failure. Second group crowns were directly cemented onto the Co‐Cr dies following the manufacturing process and loaded to fracture. Fractographic markings were analyzed through scanning electron microscope. Spearman correlation analysis, Kruskal–Wallis H test, Mann–Whitney U test, and Wilcoxon Signed Rank test were applied (α = .05). Fracture loads of Em crowns were higher than other groups (p < .05), with and without the aging procedure. Except for second group Pr (r = ?.532), no significant relationship was found between microcrack numbers and fracture loads (p > .05). Thermal cycling did not affect microcrack numbers and fracture loads (p > .05). Tooth‐shaped multilayered Pr blocks did not provide an advantage in terms of microcrack and fracture loads.     

Morphological description of the immature stages of Hermetia illucens (Linnaeus, 1758) (Diptera: Stratiomyidae)

In the dipteran genus Hermetia, only 6 of the 78 valid species have documented immature stages: H. albitarsis Fabricius, 1805, H. aurata Bellardi, 1859, H. concinna Williston, 1900, H. illucens (Linnaeus, 1758), H. panamensis Greene, 1940 and H. pulchra Weidemann, 1830. In particular, H. illucens stands out due to its reported applicability for forensic, medical and economic purposes. Here, we described the morphology of eggs and immature stages of this species, with a view to detecting differences between instars and in the pupal stage, which should eventually help properly identifying larval age. We utilized both optical and scanning electron microscopy tools. The eggs are elliptical and elongated, and color varies from cream white to yellowish. The larvae are apodal, hemichephalic and holopneustic, flattened dorso‐ventrally and may be recognized by the head elongated, dorsal and ventral chaetotaxy of the cephalic capsule, thoracic and abdominal segments, and the morphology of the anterior and posterior spiracles. The pupae are adecticous and coarctate, tegument dark brown and pruinescence varying from brown to golden. The overall morphology across instars is similar, but marked variations were observed in the shape of the antennal articuli and the shape of the setae (first instar compared to the others). Our results supplement the biological information on Hermetia illucens and should aid the proper identification and aging of juveniles in the field, as a way to minimize errors in the calculation of the post‐mortem interval.     

Observations on the oral organelle,cytopharynx, and subpellicular structure of a Dileptus sp.

We used scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to observe the oral organelle, cytopharynx, and subpellicular structure of a Dileptus sp. The main results were as follows: (a) the cytostome was located on the ventral surface of the base of the beak, surrounded by a periportal matrix that integrated 135 microtube bundles. When these microtube bundles contract, radially arranged into a disk, the cytostome was closed. When these microtube bundles were stretch, they fell into the cytostome and opens. The diameter of the cytostome was about 16 μm regardless of its closure or opening, indicating that the contraction or elongation of these microtube bundles did not change the size of the cytostome, which was only related to whether it blocked the cytostome, thus determining the opening and closing of the cytostome. There were many microtube bundles on two sides of the feeding trough, which could widen or narrow the feeding trough and facilitate beak feeding. (b) The cytopharynx was basket‐like without a bottom with a diameter of about 6 μm and was woven from two kind fibers about 0.08 and 0.19 μm. (c) There were two types of extrusomes under the pellicle. Using transmission electron microscopy，the Type I extrusomes showed narrow and long egg shape, its cross section was circular which is composed by various electronic density of concentric. Using the scanning electron microscope, they were two slightly thin clavate, the length was about 5 μm, the diameter of the middle section was about 0.75 μm, and the diameter of the two ends was about 0.32 μm, they were distributed abundantly between the microtubule fasciculi which were located on both sides of the gap on the feeding groove. Using transmission electron microscopy, the Type II extrusomes showed egg shape. Using the scanning electron microscopy, they were about 1.6 × 0.8 μm in size, they were distributed abundantly under the body pellicle while rarely the proboscis. In addition, many different of developmental stages two types of extrusomes could be also seen in the cytoplasm. (d) There were very well‐developed fibrous systems under the pellicle that were woven from fibers about 0.14 μm in diameter that attached to the pellicle and bound some organelles in the cytoplasm (e.g., mitochondria, extrusomes) and other structures to the cytoplasm and maintained cell morphology. The results of this study not only supplement and enrich the morphological contents of the Dileptus sp., but also provide the basis for the study of the taxonomy of the Dileptus sp. It also provides a new method for researchers to explore the morphology and structure of ciliate cells under the cortex by SEM.     

Propagation of transient out-of-plane shear waves in viscoelastic layered media

Propagation of two-dimensional transient out-of-plane shear waves in multilayered viscoelastic media is investigated. The multilayered medium consists of N different isotropic, homogeneous and linearly viscoelastic layers with more than one discrete relaxation time. The top surface of the layered medium is subjected to dynamic out-of-plane shear tractions; whereas, the lower surface is free or fixed. A numerical technique is employed to obtain the solution, which combines the Fourier transform with the method of characteristics. The numerical results are displayed in curves denoting the variations of the shear stresses with time at different locations. These curves reveal clearly the scattering effects caused by the reflections and refractions of inclined waves at the boundaries and at the interfaces of the layers. The curves also display the effects of viscous damping in the wave profiles. By suitably adjusting the material constants, the curves for the case of elastic layers are also obtained as a special case. The curves further show that the numerical technique applied in this study is capable of predicting the sharp variations at the wave fronts.     

Ultrastructure of oocytes of the Urostreptus atrobrunneus (Diplopoda,Spirostreptida, Spirostreptidae): A potential urban centers plague

The knowledge of the process of egg formation is indispensable for understanding the mechanisms involved in the reproduction of different species. In this context, the objective of this work was to describe the ultrastructure of the oocytes of Urostreptus atrobrunneus (Spirostreptida), a potential plague of urban centers in different locations of São Paulo State. The lack of knowledge about the morphology, physiology, and the reproductive behavior of the species have hindered an effective control of it. The oocytes of U. atrobrunneus presented three development stages: young oocyte or type I; intermediary oocyte or type II; and mature oocyte or type III. During the oocyte development, the cytoplasm become filled with several globules of protein, drops of lipids, and sphaerocrystals, and it was not observed in many organelles in the oocytes with exception of mitochondria, abundant, principally in young oocytes. The vitelline membrane is also deposited in a discontinuous form and the chorion does not present differentiation of layers. The follicular epithelium alters its shape according to the development phase of the oocyte. Part of the vitellus is from exogenous origin and part is endogenous. Before this, only two studies about the ultrastructural analysis of the female germ cells of diplopods were published. Microsc. Res. Tech. © 2012 Wiley Periodicals, Inc.     

The biomechanical,chemical and physiological adaptations of the eggs of two Australian megapodes to their nesting strategies and their implications for extinct titanosaur dinosaurs

Megapodes are galliform birds endemic to Australasia and unusual among modern birds in that they bury their eggs for incubation in diverse substrates and using various strategies. Alectura lathami and Leipoa ocellata are Australian megapodes that build and nest in mounds of soil and organic matter. Such unusual nesting behaviours have resulted in particular evolutionary adaptations of their eggs and eggshells. We used a combination of scanning electron microscopy, including electron backscatter diffraction and energy‐dispersive X‐ray spectroscopy, to determine the fine structure of the eggshells and micro‐CT scanning to map the structure of pores. We discovered that the surface of the eggshell of A. lathami displays nodes similar to those of extinct titanosaur dinosaurs from Transylvania and Auca Mahuevo egg layer #4. We propose that this pronounced nodular ornamentation is an adaptation to an environment rich in organic acids from their nest mound, protecting the egg surface from chemical etching and leaving the eggshell thickness intact. By contrast, L. ocellata nests in mounds of sand with less organic matter in semiarid environments and has eggshells with weakly defined nodes, like those of extinct titanosaurs from AM L#3 that also lived in a semiarid environment. We suggest the internode spaces in both megapode and titanosaur species act as funnels, which concentrate the condensed water vapour between the nodes. This water funnelling in megapodes through the layer of calcium phosphate reduces the likelihood of bacterial infection by creating a barrier to microbial invasion. In addition, the accessory layer of both species possesses sulphur, which reinforces the calcium phosphate barrier to bacterial and fungal contamination. Like titanosaurs, pores through the eggshell are Y‐shaped in both species, but A. lathami displays unique mid‐shell connections tangential to the eggshell surface and that connect some adjacent pores, like the eggshells of titanosaur of AM L#4 and Transylvania. The function of these interconnections is not known, but likely helps the diffusion of gases in eggs buried in environments where occlusion of pores is possible.     

HRTEM of 1DSnTe@SWNT nanocomposite located on thin layers of graphite

The method for imaging of highly sensitive nanostructures unstable under electron beam irradiation is introduced. To reduce charge and thermally generated beam damage, highly conductive multilayered graphene or thin graphite layers were used as supports for nanostructures. Well‐defined crystalline structure of graphite layers enables image reconstruction by Fourier filtering and allows maintaining high quality of images. The approach was tested for imaging of highly sensitive quasi one‐dimensional SnTe nanocrystals hosted inside single‐walled carbon nanotubes. Relying on the filtered images and the image simulation, the structure of one‐dimensional SnTe was established as a chain of fcc NaCl type unit cells, connected by the [001] edges with <110> direction coinciding with nanotube axis.     

Complementary techniques for the characterization of thin film Ti/Nb multilayers

An aberration corrector on the probe-forming lens of a scanning TEM (STEM) equipped with an electron energy-loss spectrometer (EELS) and X-ray energy-dispersive spectrometer (XEDS) has been employed to investigate the compositional variations as a function of length scale in nanoscale Ti/Nb metallic multilayers. The composition profiles of EELS and XEDS were compared with the profiles obtained from the complementary technique of 3D atom probe tomography. At large layer widths (h≥7 nm, where h is the layer width) of Ti and Nb, XEDS composition profiles of Ti/Nb metallic multilayers are in good agreement with the EELS results. However, at reduced layer widths (h≈2 nm), profiles of EELS and atom probe exhibited similar compositional variations, whereas XEDS results have shown a marked difference. This difference in the composition profiling of the layers has been addressed with reference to the effects of beam broadening and the origin of the signals collected in these techniques. The advantage of using EELS over XEDS for these nanoscaled multilayered materials is demonstrated.     

Defense reactions of <i>Dermatobia hominis</i> (Diptera: Cuterebridae) larval hemocytes

The defense reactions against biological (Histoplasma capsulatum and Escherichia coli) and non-biological materials (China ink and nylon thread) were tested in vivo in third instar larvae of Dermatobia hominis. The cellular defense performed by larval hemocytes was observed under electron microscopy. China ink particles were phagocytosed by granular cells 5 h after injection. E. coli cells were internalized by granular cells as early as 5 min after injection and totally cleared 180 min post-injection, when many hemocytes appeared disintegrated and others in process of recovering. H. capsulatum yeasts provoked, 24 h after being injected, the beginning of nodule formation. Nylon thread was encapsulated 24 h after the introduction into the hemocoel. Our results suggest that granular cells were the phagocytic cells and also the responsible for the triggering of nodule and capsule formation. In the presence of yeasts cells and nylon thread, they released their granules that chemotactically attracted the plasmatocytes that on their turn, flattened to surround and isolate the foreign material.     

Confocal microscopy of the in-situ crystalline lens

The fine structure of the in-situ rabbit crystalline ocular lens from the ex-vivo rabbit eye was observed with a confocal scanning laser microscope in the scattered light mode. The images were observed through the full thickness of the cornea and aqueous humour to a depth of 50 μm in the anterior ocular lens. The following structures were observed from optical sections of the ocular lens: two concentric regions of the lens capsule, epithelial cells, lens sutures, and surface and interior regions of individual lenticular fibres. The observed lateral resolution of the microscope objective was degraded by imaging across thick (millimetre) structures. This study shows the feasibility of obtaining high-contrast images of transparent objects across 1.7 mm of ocular tissue (cornea and aqueous humour) using confocal light microscopy.