Morphological characterization of the mouthparts of the vector leafhopper Psammotettix striatus (L.) (Hemiptera: Cicadellidae)

The leafhopper Psammotettix striatus (L.) (Hemiptera: Cicadellidae), is one of the most significant economic pests of wheat in Western China. This insect vectors a phytoplasma that causes wheat blue dwarf (WBD), a severe disease limiting wheat production in the Western China. A microscopic analysis of the ultrastructure of the mouthparts of the adult was conducted using scanning electron microscopy and the putative functions of the mouthparts were determined. The piercing-sucking mouthparts of P. striatus are of the conventional type comprising a three-segmented labium with a deep groove in the anterior side, a stylet fascicle consisting of two mandibular and two maxillary stylets, and an uppermost small cone-shaped labrum. The mandibular stylets, located laterad of the maxillary stylets, have sculpture on their tips, which may function in tearing plant tissue, cutting channels into the plant tissues, and attaching the body to the host plant during molting. The maxillary stylets are interlocked to form two separate compartments, a larger food canal and a smaller salivary canal. Two dendritic canals are also found in each maxilla and one in each mandible. Four kinds of sensilla were found on the labium: s. trichodea I, s. trichodea II, s. basiconic I, s. basiconic II. These may be involved in host recognition and are likely chemo- or mechanosensory, or both.     

The Sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae) digestive tract: histology, ultrastructure and its physiological significance

The Sunn pest, Eurygaster integriceps Put. (Hemiptera: Scutelleridae), is a key pest of wheat in the Middle East and some other areas which causes severe qualitative and quantitative damage. The objective of the current work is to describe the morphology of the midgut of E. integriceps adult. Microscopic studies revealed that foregut consists of oral cavity, pharynx and oesophagus likely other phytophagous Hemiptera. In the Midgut, four anatomical regions could be identified: the first ventriculus (V1), the second ventriculus (V2), the third ventriculs (V3), and the fourth ventriculus (V4). The microvilli and perimicrovillar membrane (PMM) were found in V1-V3 regions with columnar cells characterized by presence of mitochondria, rough endoplasmic reticulum and basal infoldings in the basal portion. However, V2 and V3 showed less developed basal plasma membrane infoldings. Three cell types: columnar, endocrine and regenerative cells were found in V1-V3. The V4 region showed different histological features from the other three midgut regions by showing a vacuolated epithelium with crypts storing symbiotic bacteria. The hindgut had a short ileum followed by a well-developed rectum with an epithelial cell layer and a thin cuticular intima. The current results suggest V1-V3 midgut regions play a role in enzyme and absorption, whereas V4 seems to have no function in digestion.     

Morphology,distribution and abundance of antennal sensilla in three stink bug species (Hemiptera: Pentatomidae)

The neotropical stink bugs, Euschistus heros, Piezodorus guildinii and Edessa meditabunda, are important pests of soybean and other crops throughout Central America and in South America from Northern Argentina to Brazil. Mate finding and host plant location in these species depend largely on their chemical communication, and semiochemicals are important mediators of these behaviors. In this study scanning electron microscopy was used to examine the external morphology, distribution and abundance of antennal sensilla on males, females and 5th instar nymphs of these species. Nine morphologically different sensilla types were found: trichod sensilla, type 1 and 2 (ST1 and ST2), long and short basiconic sensilla (SB1, SB2, and SB3), slit-tipped and knob-shaped basiconic sensilla, long chaetic sensilla (Sch) and coeloconic sensilla (Sco). Differences were detected in the abundance and arrangement of the sensilla over the antennal segments in individuals of the same species and among the species studied. The Sch, Sco and the slit-tipped and knob-shaped basiconic sensilla accounted for the major difference in sensilla types among the species. The ST1 was the most abundant type and was restricted to the flagellum. The pedicel of E. heros differs from the pedicels of P. guildinii and E. meditabunda mainly by the absence of Sch. There was a sexual dimorphism in ST1, SB1 and SB2, and this may be an indicative of their roles in detection of male-produced sex pheromone and odors derived from the host plants. The SB2 was lacking in the antennal tip of both sexes and 5th instar nymphs, but was abundant on the second flagellar segment of females of the three species. The same types of sensilla were found on 5th instar nymphs, but always in significantly lower numbers. The morphology and putative functions of each sensilla were compared and discussed.     

Ultra-structure and histochemistry of digestive cells of Podisus nigrispinus (Hemiptera: Pentatomidae) fed with prey reared on bt-cotton

The toxic effects of Bt proteins from the body of the prey/host to predators and parasitoids have been investigated as a route of impact on non-target insects of Bt crops. This research aimed to analyze ultrastructural changes and histochemistry in digestive cells of the middle region of the midgut of Podisus nigrispinus fed, since its second instar, with Spodoptera frugiperda reared on Bt cotton variety Acala90 B, which expresses the toxin Cry 1Ac, and its non-Bt isoline Acala 90. Fragments of the midgut of P. nigrispinus were analyzed by electron microscopy. For the histochemical analysis, Bromophenol Blue, Periodic Acid Schiff von Kossa, Alcian Blue pH 2.5 and Sudan Black were used. The Cry1Ac toxin of Bt cotton ingested by S. frugiperda promotes a disorganization in the perimicrovillar matrix of P. nigrispinus (third trophic level), thus generating ultrastructural changes in the digestive cells, as elongation of microvilli, presence of spherocrystals and granules of different electron densities, in addition to altering the distribution pattern of glycogen, lipids and calcium of these cells in the median region of the midgut. Thus, we conclude that species moderately susceptible to Cry1Ac toxin, such as S. frugiperda, can acquire this toxin and expose it to P. nigrispinus, which can interfere with your ability to predation.     

Morphology of the male reproductive system and spermiogenesis in Hypanthidium foveolatum (Alfken, 1930) (Hymenoptera: Apidae: Megachilinae)

The morphological aspects of male reproductive tract, spermiogenesis and spermatozoa are typical for each species and reflect its evolution, establishing a unique source of characters, which has been used to help solve phylogenetic problems. In Hypanthidium foveolatum the reproductive tract is composed of the testes comprising 28 testicular tubules, deferent ducts, seminal vesicles, accessory glands and an ejaculatory duct. The differentiation of spermatids occurs within cysts of up to 128 germ line cells each one. During the early spermatid phase, the nucleus resembles that of somatic cells. There follows a gradual chromatin condensation with an increase in nuclear electron density. In the spermatozoon, the nucleus contains heterogeneous chromatin with a loose appearance. The acrosome, shaped with the active participation of the Golgi complex, shows an electron-dense perforatorium involved by four electron-lucent acrosomal vesicle projections. The sperm tail presents an axoneme with a 9 + 9 + 2 microtubule pattern and two mitochondrial derivatives, which appear with different sizes. A dense crystalloid is formed initially in the mitochondrial matrix of the large derivative. The mitochondrial derivatives’ differentiation occurs concomitantly with an axoneme outgrowth. The centriolar adjunct is observed near the axoneme, anterior to the smaller mithocondrial derivative and exhibits an approximately triangular shape in cross-sections. Microtubules were observed around the head region and flagellar components during spermiogenesis.     

Morphostructural analysis of the male reproductive system and DNA barcoding in Balclutha brevis Lindberg 1954 (Homoptera,Cicadellidae)

Balclutha brevis Lindberg 1954 is an allochthonous leafhopper infesting an invasive grass, Pennisetum setaceum, in Sicily and in mainland Europe; therefore, this species could compete with populations of native species, thus contributing to the loss of biodiversity. Considering the ecological implications of B. brevis, investigations on all its biological aspects represent, therefore, a premise for further studies in applied sciences. Based on the lacking ultrastructural data about the reproductive systems of the Auchenorrhyncha, we carried out morphostructural investigations on the male reproductive system of B. brevis. Further, a first report of DNA barcoding analysis (amplification and sequencing of Cytochrome Oxidase I gene) has also been performed to characterize B. brevis compared to other congeneric species. From a morphological point of view, the male reproductive system of B. brevis has an organization comparable to the general anatomical features of most of the Auchenorrhyncha species; however, comparing our data with those concerning the different groups of Cicadomorpha, some considerations are discussed. As for the histological and ultrastructural investigations, our results show a secretory activity of the various examined structures, mainly in the lateral ejaculatory ducts and in the accessory glands. The latter, in particular, show morphostructural differences comparing the distal tract to the proximal one; moreover, the histochemical techniques showed the possible presence of a lipid component in the peculiar cytoplasmic granules found in the gland cells. The significance of these findings in the accessory glands is discussed. Finally, the ultrastructural features found in the seminal vesicles are different from those of the lateral ejaculatory ducts and are indicative of the different roles played by these structures in the organization of the spermatozoa bundles.     

Electrohydrodynamic (EHD) drying of rapeseed (Brassica napus L.)

The drying rate and germination parameters of rapeseeds (PF variety) treated by a high-voltage electrostatic field (HVEF) were investigated. The experiment was laid out as a factorial arrangement based on a completely randomized design with three replications. The results showed that the electrostatic field had a significant effect (P < 0.01) on decreasing rapeseed moisture content. Average drying rate for 8, 9, and 10 kV electrostatic fields over a time of 270 min increased by 1.78, 2.11, and 2.47 times, respectively, compared with that of the control. Drying rate increased with increasing voltage. Moreover, the results obtained from the germination experiment showed that the EHD had significant effects on the stemlet and radicle lengths of the rapeseed sprouts compared to that of the control. No significant effects were observed on the seed germination percentage and speed.     

Scanning electron microscopy studies of antennal sensilla of bruchid beetles,Callosobruchus chinensis (L.) and Callosobruchus maculatus (F.) (Coleoptera: Bruchidae)

The bruchid beetle, Callosobruchus chinensis (L.) and C. maculatus (F.) (Coleoptera: Bruchidae), are important stored-product insects of stored legume seeds. In this study, the external morphologies of the antennal sensilla on the antennae of both female and male adults of these two species were described using scanning electron microscopy. Antennae of both species are made up of the scape, pedicel, and nine segments of flagellomeres. Antennae of female and male C. maculatus and female of C. chinensis are serrate in shape, while those of male C. chinensis are pectinate. Eight morphological sensilla types were recorded in both sexes, including Böhm bristles (BB), two types of sensilla trichoid (ST1, ST2), sensilla chaetica (SC), two types of sensilla basiconic (SB1, SB2), grooved pegs (GP), and sensilla cavity (SCa). The number of ST1 and SB1 of the male were significantly greater than those of the female of C. chinensis, and the number of ST2 and SB1 of the male were significantly more abundant than those of the female of C. maculatus. The possible functions of the above sensilla types are discussed in light of previously published literature.     

Morphology of the male reproductive system and spermatozoa in Centris Fabricius, 1804 (Hymenoptera: Apidae, Centridini)

The genera Centris and Epicharis constitute the Centridini and are widely distributed in the Neotropical region. Centris is also found in the southern portion of the Neartic region, although both genera are more abundant in the humid tropical regions. To describe the structure of the male reproductive system and spermatozoa, light and transmission electron microscopy were used. The male reproductive system of Centris sp. is formed by a pair of testes, a pair of deferent ducts, a pair of seminal vesicles, a pair of accessory glands and an ejaculatory duct connected to the external genitalia, the aedeagus. In this species, testes and the pre-vesicular deferens ducts as well as the seminal vesicles are encapsulated in a single conjunctive capsule, the scrotal membrane. Each testis consists of four testicular follicles, made up of cysts with up to 64 germinative cells. Histologically, the seminal vesicles are formed by a simple cylindrical epithelium, basal membrane and muscular tunic. The spermatozoa of Centris analis, C. fuscata, C. tarsata and Centris sp. are morphologically similar. They have two easily distinguishable regions: the head and flagellum. The head is formed by the two-layer acrosome, the linear nucleus and the flagellum, the centriole adjunct, the axoneme of pattern 9+9+2 microtubules, two asymmetric mitochondrial derivatives and two accessory bodies. These Centris species share various morphological characteristics of the male reproductive system and spermatozoa with the other bees previously described, indicating that several characteristics are synapomorphic for the family Apidae. Studies on the morphology of the male reproductive system and spermatozoa in Hymenoptera have demonstrated the diversity of the information provided by these reproductive structures, which can be used in taxonomy studies and the phylogeny of this important group of insects.     

Anatomical structure and ultrastructure of the endocarp cell walls of Argania spinosa (L.) Skeels (Sapotaceae)

The anatomical and histochemical study of young and adult endocarps of Argania spinosa (sampled from Tindouf; Algeria) shows a general structure that is similar to that of majority of stone fruits. These samples consist of tissues that contain lignified and cellulosic cell walls. The majority of the tissues are composed of sclerenchyma cells; with very thick lignified cell walls and conducting tissues. Coniferyl lignins are abundant in the majority of the lignified tissues. However, the coniferyl lignins appear at the primary xylem during lignification. Syringyl lignins are present in small quantities. The electron microscopy observation of the sclerenchyma cell walls of the young endocarp shows polylamellate strates and, cellular microfibrils in arced patterns. This architecture is observed in the cell walls of the adult endocarp only after the incubation of the tissue in methylamine. These configurations (arcs) are the result of a regular and complete rotation with a 180° variation in the microfibril angle; the complete and symmetrical arcs show a helicoidal mode of construction. The observation of the sclerenchyma cells revealed the capacity of helicoidal morphogenesis to adjust itself under the influence of topological constraints, such as the presence of a large number of pit canals, which maintain symplastic transport.     

Optical illusions in scanning electron micrographs: the case of the eggshell of Acrosternum (Chinavia) marginatum (Hemiptera: Pentatomidae)

Scanning electron microscopy revealed that-as is common in this family of the Hemiptera-the eggs of the green stink bug Acrosternum (Chinavia) marginatum are roughly barrel-shaped and possess at their apical pole a row of slender extensions, the aero-micropylar processes. The outer surface of the eggshell carries hexagonally arranged pits. The analysis of cross-fractured eggshells showed that the pits have slender basal extensions with transverse diaphragms. When scanning electron micrographs of the egg surface of A. marginatum are viewed upside down, the perception flips and the pits appear as elevations to all observers addressed. Thus, we are dealing with an optical illusion, which is known as the 'shape-from-shading effect'. The perceived dents remain robust to changes in the angle of recording (zero to ca. 60 degrees tilt), the magnification (ca. x100 to x1400), and the number of pits included in the micrograph (one to several hundred). When through appropriate positioning of the specimen under the electron beam, contrast is significantly reduced and the distinct shadows at the slope of the pits are eliminated, the optical illusion does not appear. It is inferred that shades provide the decisive clue that determines whether bumps or dents will be perceived. Owing to the low resolution of their compound eyes, the shape-from-shading effect on the eggshell of the bug is in all likelihood not perceived by insects.     

Kinetics of ultrasonic extraction of extractive substances from garden (Salvia officinalis L.) and glutinous (Salvia glutinosa L.) sage

The kinetics of ultrasonic extraction of extractive substances (ES) from dry herbs of garden (Salvia officinalis L.) and glutinous (Salvia glutinosa L.) sage using petroleum ether, 70% ethanol or water at 40 degrees C, as well as the composition of dry extracts, were studied. The mechanism of ultrasonic extraction is confirmed to occur in two steps: first, dissolution of the ES near the particle surface (washing) and, second, diffusion from the solid particles to the bulk of the liquid extract (slow extraction). The process is described mathematically using three concepts of the unsteady diffusion through plant material, the film theory and the empirical equation of Ponomaryov. The yield of ES increases with increasing solvent polarity, and nearly the maximum concentration of ES in liquid extracts is achieved for about 20 min. The composition of extracts depends on both the extraction conditions applied and the plant material.     

Ultrastructural characterization of the spermatozoa of Aethalion reticulatum Linnaeus 1767 (Hemiptera: Auchenorrhyncha: Aethalionidae)

The Hemiptera order is currently divided into four suborders. Among them the Auchenorrhyncha suborder is considered to be paraphyletic. Morphology of insect spermatozoa has provided promising characteristics that can be used for phylogenetic inference. In this study, Aethalion reticulatum (Aethalionidae) spermatozoa were examined by light and electron microscopy. The head of the spermatozoa is composed of an acrosome and a nucleus. The nucleus is linear and filled with compact chromatin but has electron-lucid spaces. The centriole adjunct initiates parallel to the nucleus and terminates in the region anterior to the mitochondrial derivatives. Flagella consist of an axoneme, two mitochondrial derivatives and two accessory bodies and the axoneme has the typical 9 + 9 + 2 microtubule pattern. The mitochondrial derivatives are symmetric. The accessory bodies are long and are shaped like a half moon when viewed from a cross-section. The presence of accessory bodies differs from other species of Cicadomorpha previously studied. Spermatozoa morphology of other Auchenorrhyncha families can reveal synapomorphies and contribute to systematics of the suborder.     

Micromorphology of cactus-pear (Opuntia ficus-indica (L.) Mill) cladodes based on scanning microscopies

Cladode ultrastructural features of two prickly and two spineless Opuntia ficus-indica cultivars were examined using environmental scanning electron and atomic force microscopies. Observations focused on cladode as well as spine and glochid surface micromorphologies. Prickly cultivars were characterized by abundant cracked epicuticular wax deposits covering the cladode surface, with an amorphous structure as observed by AFM, while less abundant waxy plates were observed by ESEM on spineless cultivar cladodes. Further AFM observations allowed a rough granular and crystalloid epicuticular wax structure to be distinguished in spineless cultivars. Regarding spine micromorphology, prickly cultivars had strong persistent spines, observed by ESEM as a compact arrangement of oblong epidermal cells with a rough granular structure. However, deciduous spines in spineless cultivars had a broken transversely fissured epidermis covering a parallel arrangement of fibres. Through AFM, the deciduous spine surface presented an irregular hilly and smooth microrelief while persistent spines exhibited rough helical filamentous prints.ESEM and AFM studies of cladode surfaces from prickly and spineless cactus pear cultivars revealed valuable micro-morphological details that ought to be extended to a large number of O. ficus-indica cultivars.     

Male reproductive system structure and accessory glands ultrastructure of two species of Triatoma (Hemiptera,Reduviidae, Triatominae)

Morphological studies in male reproductive system of insect, and accessory glands functions, can contribute to the knowledge of reproductive aspects in Triatominae. The present study aims to describe the structure of male reproductive system under scanning electron microscopy, and the ultrastructure of accessory glands during the process of maturation, in T. brasiliensis and T. melanica of different ages.These system consist of two testis, two vas deferens, two seminal vesicles, four pair of accessory glands and one ejaculatory duct. The four mesodermal male accessory glands of T. melanica and T. brasiliensis, present secretion storage in the firsts days of adult life in starved males. During the maturation process of male accessory glands of T. brasiliensis and T. melanica, granules with different sizes, shapes and electron density were seen. In T. melanica small vacuoles are released into the gland lumen in one-day-old males, however, after three and five days few granules are found in the cell cytoplasm. The secretory granules in T. brasiliensis increase in size and amount, in the five days old adult. The secretory pathway in the male accessory glands of the two species seems to be merocrine and apocrine suggesting that different substances are being produced at different times. The cell microvilli in T. brasiliensis are longer and narrower, these can be absent in some regions of the cell surface of T. melanica. Maturation of male accessory glands in the triatomine species studied occurred without adult blood meal. Its function can led to new insights for the comprehension of reproductive aspects in Triatominae.     

Ultrastructural study of the ovary of the sugarcane spittlebug Mahanarva fimbriolata (Hemiptera)

The present study describes the ultrastructure of meroistic telotrophic ovaries of the sugarcane spittlebug Mahanarva fimbriolata. In this type of ovary, nurse cells, oogonia, and prefollicular tissue are located at the terminal (distal) regions or tropharium of ovarioles. Oocytes in different developmental stages, classified from I to V, are observed in the vitellarium. Stage I oocytes do not exhibit intercellular spaces in the follicular epithelium, suggesting that synthesis and production of yolk during this stage occurs only through endogenous processes. Small yolk granules of different electron densities are present in the cytoplasm. Few lipid droplets are observed. Stage II oocytes exhibit small intercellular spaces in the follicular epithelium. More protein as well as lipid yolk granules are observed in the cytoplasm. In stage III oocytes, intercellular spaces in the follicular epithelium are larger than those observed in the previous stage. Electrondense protein granules of various sizes, larger than those observed in stage II oocytes predominate in the cytoplasm. Smaller lipid droplets are also present. In stage IV oocytes, the follicular epithelium exhibits large intercellular spaces. Our data clearly indicate that the opening of these spaces in the follicular epithelium of M. fimbriolata oocytes increases as the intake of exogenous proteins intensifies, that is, in stages IV and V oocytes. During these stages, granular yolk becomes viscous due to the lysis of granules. In stage V oocytes, viscous yolk predominates in the cytoplasm. This type of yolk, however, has not been described for other orders of insects. The chorion of M. fimbriolata oocytes consists of an external layer (exochorion) and an internal one (endochorion), which is in direct contact with the oocyte. Numerous small pores that probably facilitate oxygenation of the internal structures inside the eggs are observed in the exochorion.     

Phagocytosis of latex beads and bacteria by hemocytes of the triatomine bug Rhodnius prolixus (Hemiptera: Reduvidae)

Insect circulating hemocytes are primarily responsible for the immune defense against parasites and pathogens. Here, we have analyzed phagocytosis of both biotic (bacteria) and abiotic (latex) particles by circulating hemocytes of 5th-instar nymphs of the triatomine bug Rhodnius prolixus. The following hemocyte types were identified: prohemocytes, plasmatocytes, granulocytes, oenocytoids and adipohemocytes. There was a considerable change in the relative percentage of plasmatocytes and prohemocytes in the hemolymph after challenge with both latex beads and bacteria. Granulocytes and oenocytoids also change their relative percentage in response to latex bead and Staphylococcus aureus, respectively. No significant change was observed in adipohemocytes at any time or treatment. Our data demonstrated that plasmatocytes were the only cell type involved in phagocytosis of foreign particles. As in mammal cells, phagocytosis by both zipper and trigger mechanisms were observed for the uptake of latex beads and bacteria. Neither melanization nor micro-aggregation was observed towards latex particles or Escherichia coli. On the other hand, R. prolixus produced a strong melanization reaction against S. aureus, thus showing that differences exist in the responses to E. coli and to S. aureus. Ultrastructural changes observed in plasmatocytes, adipohemocytes and oenocytoids suggest that these hemocyte types are directly involved in the immune defense of R. prolixus against foreign particles.     

Generalised vector dominance: Off-diagonal terms and asymptotic SU(4)

Asymptotic SU(4) symmetry is imposed on generalised vector dominance models with off-diagonal terms, relating the magnitudes of these terms in the various sectors V = ?, ω, φ, ψ. While the structure functions are modified, their ratios remain similar to those of the diagonal model. Strangeness and charm production are suppressed for low Q² < M_φ², M_ψ² respectively.     

Moisture gradient vector calculation as a new method for evaluating NMR images of corn (Zea Mays L.) kernels during drying.

Magnetic resonance imaging (MRI) was used to study the moisture migration inside single corn kernel during artificial drying. MRI data were taken every half an hour during drying while the intact kernel was kept inside the magnet. The moisture distribution was not uniform inside the kernel during drying, with the fastest moisture loss being detected in the endosperm, whilst the scutellum maintained the water. Resultant gradient vectors were visualized, as a new evaluation method, from each pixel of the subtraction of segmented MRI images showing the movement of proton density inside the kernel. The data matrix containing the gradient vectors can be stored for latter use as input parameters and to check the mass transfer models.