Sperm ultrastructure and spermiogenesis of Coniopterygidae (Neuroptera, Insecta)

The spermiogenesis and the sperm ultrastructure of several species of Coniopterygidae have been examined. The spermatozoa consist of a three-layered acrosome, an elongated elliptical nucleus, a long flagellum provided with a 9+9+3 axoneme and two mitochondrial derivatives. No accessory bodies were observed. The axoneme exhibits accessory microtubules provided with 13, rather than 16, protofilaments in their tubular wall; the intertubular material is reduced and distributed differently from that observed in other Neuropterida. Sperm axoneme organization supports the isolated position of the family previously proposed on the basis of morphological data.     

Sperm ultrastructure in Chironomoidea (Insecta, Diptera)

The fine structure of spermatozoa from several species of chironomids, of Culicoides sp. (Ceratopogonidae) and of Odagmia pontina (Simulidae) was studied. A synapomorphic feature, consisting of nine kidney-shaped structures forming the centriole adjunct, was found in the chironomid species. All members of Chironomoidea share a mono-layered acrosome and a flagellar axoneme, provided with accessory tubules with 15 protofilaments in their tubular wall. The axoneme has a 9+9+2 pattern, but in an unidentified species of chironomid, a 9+9+0 model was observed where the central complex and the spokes are missing. Sperm motility is, however, maintained in all the examined species. The spermatozoa of this taxon have the tendency to complete maturation during their progression along the deferent ducts. Thus, in the proximal region of these ducts, they often show remnants of the spermatid cytoplasm.     

On Afromantispa and Mantispa (Insecta,Neuroptera, Mantispidae): elucidating generic boundaries

The genus Afromantispa Snyman & Ohl, 2012 was recently synonymised with Mantispa Illiger, 1798 by Monserrat (2014). Here morphological evidence is presented in support of restoring the genus Afromantispa stat. rev. to its previous status as a valid and morphologically distinct genus. Twelve new combinations (comb. n.) are proposed as species of Afromantispa including three new synonyms.     

The spermatogenesis and the sperm structure of Terebrantia (Thysanoptera, Insecta)

Spermatogenesis and the sperm structure of the terebrantian Aeolothrips intermedius Bagnall are described. Spermatogenesis consists of two mitotic divisions; the second is characterized by the loss of half of the spermatids, which have pyknotic nuclei. Early spermatids have two centrioles, but when spermiogenesis starts, a third centriole is produced. The three basal bodies give rise to three flagella; later these fuse into a single flagellum which contains three 9 + 0 axonemes. The basal bodies are surrounded by a large amount of centriole adjunct material. During spermiogenesis this material contributes to the shifting of the three axonemes towards the anterior sperm region parallel to the elongating nucleus, and it is transformed into a dense cylinder. In the mature spermatids the three axonemes amalgamate to create a bundle of 27 doublet microtubules. Near the end of spermiogenesis the dense cylinder of the centriole adjunct lies parallel to the nucleus and the axonemes. It ends where the mitochondrion appears at half-sperm length. We confirm that Terebrantia testes have a single sperm cyst; their sperm are characterized by a cylindrical nucleus, three axonemes fused into one, a small mitochondrion and a short cylindrical centriole adjunct which corresponds to the dense body described in a previous work. The acrosome is lacking. At the midpoint of the anterior half of the sperm the outline of the cross-section is bilobed, with the nucleus contained in a pocket evagination of the plasma membrane. These characters are discussed in light of a comparison between Tubulifera and Terebrantia.     

The spermatodesm of Cloeon dipterum (L.): fine structure and sperm movement

Sperm bundles of the ephemeropteran Cloeon dipterum (L.) were found in the seminal vesicle. They are apically protected by cyst cells and sperm anterior regions are embedded in a secretion produced by these cells. Cyst cells further degenerate making sperm cells free. The sperm bundles are able to perform a forward progression only when sperm are stick together with their tail posterior tips. On the contrary, when sperm have their tail ends free, they beat but are unable to progress. The functional significance of the sperm bundles is discussed.     

Morphology of the male reproductive system and sperm ultrastructure of the egg parasitoid Gryon pennsylvanicum (Ashmead) (Hymenoptera,Platygastridae)

Gryon pennsylvanicum is a platygastrid hymenopteran that has lately received increasing attention in Europe due to its possible use in biological control of the conifer seed bug pest Leptoglossus occidentalis. Here the male reproductive system and the spermatogenesis of this species, along with those of Gryon muscaeformis, are examined ultrastructurally for the first time. The male genital system is formed by a pair of testes, each containing only one follicle, a pair of accessory glands and deferent ducts connected to a single ejaculatory duct. All the stages of spermatogenesis are described in detail. Characteristic features of the Gryon spp. sperm, which are 100 μm long, are the presence of a polygonal nucleus, only one mitochondrial derivative, the occurrence of the centriole adjunct and a typical insect 9 + 9 + 2 flagellar axoneme. The single derivative, however, results from a process in which one of the two mitochondria is lost during spermiogenesis. Unlike in other insects, two centrioles occur in spermatids as a consequence of the ameiotic parthenogenesis. These characteristics stand as a valuable tool for phylogenetic inferences. Furthermore this study suggests a useful strategy for laboratory mass rearing.     

The male reproductive system of Zorotypus caudelli Karny (Zoraptera): Sperm structure and spermiogenesis

Considering the overall uniformity of the morphology of Zoraptera, the structural diversity of the male genital system is remarkable. Structures related to the male reproductive system of Zorotypus caudelli differ profoundly from those of Zorotypus hubbardi. The testes are elongated rather than spherical, the seminal vesicle is apparently absent, and the deferent ducts are very long. A feature shared by these two species and other zorapterans examined is that the two accessory glands are closely adherent to each other and form a single large structure, from which the ejaculatory duct originates. This is a potential zorapteran autapomorphy. Another feature possibly present in the groundplan of the order is the strong elongation of the sperm cells. This may be connected with a reproductive strategy of males trying to avoid re-mating of females with other males after the first copulation. The extremely long and coiled spermathecal duct of Z. caudelli and other zorapteran species is possibly correlated with the sperm elongation, and both features combined may result in a sexual isolating mechanism. The short duration of mating of Zorotypus barberi and Zorotypus gurneyi suggests that the male introduces sperm into the female tract up to the opening of the spermathecal duct using their long coiled aedeagus. A thick glycocalyx around the sperm in the distal part of the deferent ducts probably protects the sperm cells during their forward progression towards the long spermathecal duct, and is removed when they reach the apical receptacle. The spermatogenesis of Z. caudelli follows a pattern commonly found in insects, but differs distinctly from that of Z. hubbardi in the number of spermatids in each sperm cyst. An unusual and possibly autapomorphic feature of Z. caudelli is a disconnection of sub-tubules A and B at the level of microtubule doublets 1 and 6 of the mature sperm cells. It is conceivable that this results in a shorter period of sperm motility. The character combination found in different zorapteran species supports the view that the sperm, a very compact functional unit, does not evolve as a unit, but like in other more complex body regions, sperm components can also be modified independently from each other. This results in different mosaic patterns of plesiomorphic and derived features in a very compact entity in different species of the very small and otherwise uniform order Zoraptera. In Z. caudelli, for instance, the bi-layered acrosome and small accessory bodies are plesiomorphic states among several others, whereas the mitochondrial derivatives and the elongate nucleus are apparently derived conditions. Other combinations likely occur in other zorapteran species. Only few but noteworthy sperm characters indicate possible phylogenetic affinities of Zoraptera. A possible synapomorphic feature, the presence of dense laminae radiating in a cartwheel array between neighbouring centriolar triplets, is shared with Phasmatodea and Embioptera. Another potential synapomorphy shared with Phasmatodea is the presence of 17 protofilaments in the tubular wall of the outer accessory microtubules.     

Specialization of the endoplasmic reticulum in the apex of the midgut cells of Aeshna cyanea (Insecta,Odonata)

Summary Lamellar stacks of cisternae regularly spaced and bound to each other by fine filaments are observed in the apex of midgut cells of Aeshna cyanea. These cisternae are connected with the rough endoplasmic reticulum but are devoid of ribosomes except on their terminal sacs. Their location and their structure, quite different in fed and starved animals, suggest that they are involved in intracellular transport.     

The fine structure of the female reproductive system of Zorotypus caudelli Karny (Zoraptera)

The general structure of the female genital system of Zorotypus caudelli is described. The ovarioles are of the panoistic type. Due to the reduction of the envelope (tunica externa) the ovarioles are in direct contact with the hemolymph like in some other insect groups, Plecoptera included. The calices are much larger in Z. caudelli then in Zorotypus hubbardi and their epithelial cells produce large amounts of secretions, probably protecting the surface of the eggs deposited on the substrate. Eggs taken from the calyx bear a series of long fringes, which are missing in the eggs found in the ovariole, and in other zorapteran species. The long sperm of Z. caudelli and the long spermathecal duct are likely related to a sexual isolating mechanism (cryptic female choice), impeding female re-mating. The apical receptacle and the spermathecal duct - both of ectodermal origin - consist of three cell types. In addition to the cells beneath the cuticle lining the lumen, two other cell types are visible: secretory and canal cells. The cytoplasm of the former is rich in rough endoplasmic reticulum cisterns and Golgi complexes, which produce numerous discrete dense secretory bodies. These products are released into the receiving canal crossing the extracellular cavity of secretory cells, extending over a series of long microvilli. The secretion is transported towards the lumen of the apical receptacle of the spermatheca or to that of the spermathecal duct by a connecting canal formed by the canal cells. It is enriched by material produced by the slender canal cells. Before mating, the sperm cells are enveloped by a thick glycocalyx produced at the level of the male accessory glands, but it is absent when they have reached the apical receptacle, and also in the spermathecal duct lumen. It is likely removed by secretions of the spermatheca. The eggs are fertilized at the level of the common oviduct where the spermathecal duct opens. Two micropyles at the dorsal side of the equator level possibly facilitate fertilization. The presence of these two micropyles is a presumably derived feature shared with Phasmatodea. The fine structure of the female reproductive system of Z. caudelli does not allow to assess the phylogenetic position at the present stage of knowledge. The enlarged calyx and the temporary presence of long fringes on the eggs are potential autapomorphies of Z. caudelli or may indicate relationships with other Zorotypus species.     

New findings on sperm ultrastructure of Xenos vesparum (Rossi) (Strepsiptera,Insecta)

The systematic position of insect order Strepsiptera is still under debate. It was, therefore, thought of interest to examine the ultrastructure of a strepsipteran in a search for synapomorphies shared with Coleoptera, Diptera, or any other insect order. The fine structure of spermatozoa and the spermatid from Xenos vesparum (Rossi) was re-examined using scanning and transmission electron microscopy and a fixation technique that permits the visualization of the macromolecular organization of the organelles. The spermatozoon was shown to possess several traits that are characteristics of insects in general, such as a 9 + 9 + 2 axoneme, two mitochondrial derivatives containing a crystalline material and two 'zipper lines' present along the sperm tail. Seventeen protofilaments occurred along most of the accessory tubules, which reduced to 16 posteriorly. An acrosome is absent. The neck region contains a prominent centriolar adjunct, which gives rise to two accessory bodies which adhere to the mitochondrial derivatives, and to slender strands of the so-called intertubular material found between the accessory tubules. Of interest is the finding that the glycocalyx consists of prominent filamentous strands, similar to those found in siphonapterans, mecopterans and basal dipterans.     

Ultrastructure of euspermatozoa and paraspermatozoa in the marine gastropod Adelomelon beckii (Caenogastropoda, Volutidae)

The sperm morphology of Adelomelon beckii is described by optical and transmission electron microscopy. Both euspermatozoa and paraspermatozoa were found in the specimens studied. Euspermatozoa are filiform and have an elongate nucleus capped by an acrosome. A small basal plate lies between the base of the acrosome and the nucleus. The mid-piece consists of U-shaped mitochondria wrapped helically around the central axoneme. A dense annulus at the junction of the mid-piece and glycogen piece is found, ending in a short end-piece, composed of the axoneme surrounded by a plasma membrane. Two types of paraspermatozoa are found, both vermiform but differing internally with respect to the disposition and number of axonemes, as well as to the types of secretory vesicles. We suggest the use of paraspermatozoa as a systematic character to reveal phylogenetic relationships in this family.     

Ultrastructural studies of the termite (Odontotermes obesus) gut microflora and its cellulolytic properties

The major gut microflora colonizing the hind gut of a higher termite,Odontotermes obesus, included morphologically diverse bacteria, both coccoid and rod-shaped, along with spirochaetes, pseudomonads and actinomycetes. Flagellated protozoa were totally absent. When the gut extract was inoculated on plates containing carboxymethyl cellulose or cellobiose, higher numbers of bacteria grew than on plates without cellulosic sources. The gut homogenate exhibited strong hydrolytic activity when carboxymethyl cellulose,p-nitrophenyl--d-glucoside or xylan were used as substrate, indicating the role of gut microbiota in the process of cellulose and hemicellulose digestion. Activities were highest in the hind gut, and the paunch was probably the major site of polysaccharide digestion in this higher termite.In vitro cultivation of some of the isolates revealed both cellulase and xylanase activities. To our knowledge, this is the first report on ultrastructural studies of the higher termiteOdontotermes obesus.     

Comparative morphology of spermatozoa and reproductive systems of zorapteran species from different world regions (Insecta,Zoraptera)

The male and female reproductive apparatus of Zorotypus magnicaudelli (Malaysia), Zorotypus huxleyi (Ecuador) and Zorotypus weidneri (Brazil) were examined and documented in detail. The genital apparatus and sperm of the three species show only minor differences. The testes are larger in Z. magnicaudelli. Z. huxleyi lacks the helical appendage in the accessory glands. A long cuticular flagellum is present in Z. magnicaudelli and in the previously studied Zorotypus caudelli like in several other species, whereas it is absent in Z. weidneri, Z. huxleyi, Zorotypus hubbardi, Zorotypus impolitus and Zorotypus guineensis. Characteristic features of the very similar sperm are the presence of: a) two dense arches above the axoneme; b) a 9 + 9+2 axoneme with detached subtubules A and B of doublets 1 and 6; c) the axonemal end degenerating with enlarging accessory tubules; d) accessory tubules with 17 protofilaments; e) three accessory bodies beneath the axoneme; and f) two mitochondrial derivatives of equal shape. The first characteristic (a) is unknown outside of Zoraptera and possibly autapomorphic. The sperm structure differs distinctly in Z. impolitus and Z. hubbardi, which produce giant sperm and possess a huge spermatheca. The presence of the same sperm type in species either provided with a sclerotized coiled flagellum in males or lacking this structure indicates that a different organization of the genital apparatus does not necessarily affect the sperm structure. The flagellum and its pouch has probably evolved within Zoraptera, but it cannot be excluded that it is a groundplan feature and was reduced several times. The fossil evidence and our findings suggest that distinct modifications in the genital apparatus occurred before the fragmentation of the Gondwanan landmass in the middle Cretaceous.     

Ultrastructural and phylogenetic studies on Blastocystis isolates from cockroaches

Four Blastocystis isolates from cockroaches were established and these isolates were morphologically confirmed as Blastocystis organisms by light and/or electron microscopy. As these isolates were morphologically indistinguishable from Blastocystis isolated from other animals, phylogenetic analyses were conducted using their small subunit ribosomal RNA genes. A analyses of these sequences with previously reported ones that had been classified into nine Blastocystis clades indicated the presence of a new clade that comprised only Blastocystis organisms from cockroaches (clade X). A clade comprised of amphibian and reptilian Blastocystis organisms (clade IX) was located at the basal position of the Blastocystis tree together with the common ancestor of Proteromonas and Protoopalina, clade X emerged after the divergences of these two basal clades and its branching position was clearly supported by bootstrap analysis.     

An enigmatic new stick insect from the Philippine Islands (Insecta: Phasmatodea)

A new genus and species of stick insect is described and figured from Mount Halcon, on the Philippine island of Mindoro. Conlephasma enigma gen. et sp. n. is a stout, flightless, and apparently ground-dwelling species with vivid integumental colors. When disturbed, specimens spray a defensive secretion from the prothoracic exocrine glands. The systematic position of Conlephasma within Euphasmatodea is unclear. The elongated galealobulus and the trichome area located laterally in the galea, represent unusual apomorphic characters of the maxilla that could indicate affinities with Necrosciinae or Pseudophasmatinae. All tibiae exhibit the anareolate condition. Euplantulae are of two types: those of tarsomeres I–IV feature a nubby microstructure, whilst the one on the ventral side of the pretarsus is smooth. Males are characterized by the presence of a well-developed vomer on the tenth abdominal segment. A distinctive and apomorphic trait of female terminalia is represented by the elongated tenth abdominal tergum. Conlephasma can represent an interesting taxon for studies on the evolution of the stick and leaf insects.     

Distance segregation and compound sex chromosomes in mantispids (Neuroptera: Mantispidae)

The sex chromosomes segregate precociously in prometaphase I of male meiosis, without prior synapsis or any physical connection, in 4 species of American mantispids (Neuroptera: Mantispidae). Segregational movements are interpolar, and are implemented through chromosomal fibers. Univalent autosomes, present from diakinesis on in several species, are capable of a similar distance segregation in prometaphase. The sex chromosomes are XX —XY , as is characteristic of the Order, with the exception of Entanoneura phthisica in which both elements are compound —X¹X²X³Y¹Y²Y³in the male, and X¹X¹X²X²X³X³ in the female. In tetraploid sectors of gonial origin in testes of this species no sex bivalents are formed; a distance segregation of 6 sex univalents to each pole is effected, but — as observed in the one individually identifiable pair — segregation separates complete homologues, Y¹ from Y¹, X¹ from X¹, etc. In all species the male meiotic spindle is formed by the collocation of individual chromosomal spindle units within which bivalents become deformed; the timing and degree of deformation vary with the species. In karyotype the American species conform to a common pattern with the known Japanese and European species; diploid numbers range only from 18 to 22, and each complement carries the family insigné of one pair of disproportionately large autosomes in a set of small and rather uniformly sized chromosomes.     

Chromosome numbers in antlions (Myrmeleontidae) and owlflies (Ascalaphidae) (Insecta,Neuroptera)

A short review of main cytogenetic features of insects belonging to the sister neuropteran families Myrmeleontidae (antlions) and Ascalaphidae (owlflies) is presented, with a particular focus on their chromosome numbers and sex chromosome systems. Diploid male chromosome numbers are listed for 37 species, 21 genera from 9 subfamilies of the antlions as well as for seven species and five genera of the owlfly subfamily Ascalaphinae. The list includes data on five species whose karyotypes were studied in the present work. It is shown here that antlions and owlflies share a simple sex chromosome system XY/XX; a similar range of chromosome numbers, 2n = 14-26 and 2n = 18-22 respectively; and a peculiar distant pairing of sex chromosomes in male meiosis. Usually the karyotype is particularly stable within a genus but there are some exceptions in both families (in the genera Palpares and Libelloides respectively). The Myrmeleontidae and Ascalaphidae differ in their modal chromosome numbers. Most antlions exhibit 2n = 14 and 16, and Palparinae are the only subfamily characterized by higher numbers, 2n = 22, 24, and 26. The higher numbers, 2n = 20 and 22, are also found in owlflies. Since the Palparinae represent a basal phylogenetic lineage of the Myrmeleontidae, it is hypothesized that higher chromosome numbers are ancestral for antlions and were inherited from the common ancestor of Myrmeleontidae + Ascalaphidae. They were preserved in the Palparinae (Myrmeleontidae), but changed via chromosomal fusions toward lower numbers in other subfamilies.     

Ultrastructural and cytochemical studies on the cytodifferentiation of duodenal endocrine cells

Summary The development and cytodifferentiation of endocrine cells that produce the gastrointestinal hormones gastrin, cholecystokinin and secretin have been studied by a combined fluorescence-cytochemical, immunocytochemical and ultrastructural approach. The results show that, during development, several ultrastructurally distinct cell types exhibit COOH-terminal gastrin and cholecystokinin immunoreactivity. Furthermore, some cells simultaneously contain both gastrin- and cholecystokinin-specific antigenic determinants. Studies on the time course of development of gastrin and cholecystokinin cells, together with the above-mentioned data, suggest that gastrin cells may be converted into cholecystokinin cells in development. During this period, gastrin, cholecystokinin and secretin cells store the biogenic monoamine, 5-hydroxytryptamine a feature not displayed by the adult counter-parts of these cells. In the adult duodenum, characteristic enterochromaffin (EC) cells store 5-hydroxytryptamin for which, evidence for a possible hormonal role has been presented. Taken together, our data indicate that the differentiation of duodenal endocrine cells occurs in distinct steps, each involving a restriction in the biosynthetic repertoire of the cell.