Sperm structure of the diving beetle Deronectes moestus incospectus (Leprieur, 1876) (Hydroporinae, Dytiscidae) and considerations on extracellular material surrounding sperm bundles.

The sperm cells of the diving beetle Deronectes moestus incospectus are characterized by sperm conjugation leading to the formation of sperm bundles of 64 units each. These bundles are formed at the end of spermatocyte cell divisions occurring in the testes and can be detected in the anterior region of the deferent ducts (first type of sperm conjugation). Fusions of some sperm bundles can occur at the end of the deferent ducts. The sperm bundles show sperm-head stacks (sperm rouleaux) and are surrounded by a cup of extracellular material secreted by the epithelial cells of the deferent ducts. This material extends posteriorly around the sperm bundle to cover the nuclei and the initial region of the sperm flagella. The cup extracellular material consists of fine tubules, and is no longer visible in sperm bundles at the posterior end of the deferent ducts. The sperm cells of D. moestus incospectus have an axoneme with a 9 + 9 + 2 pattern and unusual mitochondrial derivatives having a matrix showing dense dots and a small crystallized domain. Two thin elongated accessory bodies are located between the mitochondrial derivatives and the axoneme. The extracellular material can have different morphologies in the various families of Adephaga, but all are produced by the epithelium of the deferent ducts. Thus it is reasonable to assume that it has the same function in the different groups.

Fine structure of the female genital system of diving beetle Stictonectes optatus (Seidlitz, 1887) (Dytiscidae-Hydroporinae) and evidence of mating plug formation.

The general organization of the female genital system of the diving beetle Stictonectes optatus was studied, clarifying the complex structure of the spermatheca and spermathecal gland. The two structures adhere closely to each other, sharing a small area of their cuticular epithelium. A long duct connects the bursa copulatrix to the spermatheca, where the sperm are stored. The sperm reach the common oviduct, where egg fertilization occurs, via a fertilization duct. The spermathecal gland cells have extracellular cisterns where secretions are stored. Thin ducts composed of duct-forming cells transport these secretions to the apical gland region and into the spermathecal lumen. Soon after mating, the bursa copulatrix is almost completely occupied by a plug secreted by the male accessory glands. The secretions of the bursa epithelium seem to contribute to plug formation. Later this plug becomes large and spherical, obstructing the bursa copulatrix.

The sperm structure of the diving beetles Stictonectes optatus (Seidlitz, 1887) and Scarodytes halensis (Fabricius, 1787) (Dytiscidae, Hydroporinae) with evidence of a spermatostyle in the sperm conjugation.

The structure of the male genital organs and spermiogenesis of two diving beetles, Stictonectes optatus and Scarodytes halensis were studied for the first time. S. optatus shows unifollicular testes consisting of a long tubule apically forming a globular structure. The deferent duct epithelia show a secretory activity involved in the spermatostyle organization. They are connected with two very large accessory glands. Sc. halensis has a more common structure of the male genital apparatus with unifollicular cylindrical testes and very long deferent ducts. Sc. halensis accessory glands are smaller than those of S. optatus. The sperm structure in both species is characterized by a small acrosome, a flattened nucleus with a lateral extension containing a centriole from which a long flagellum originates. Both species exhibit sperm conjugation with long sperm bundles showing nuclei orderly arranged in sperm-heads stacks and free flagella. In addition, S. optatus has a thick layer of secretion surrounds these sperm-head stacks. Such a secretion is considered a spermatostyle. This finding represents the first record about the presence of this structure among Dytiscidae. In the flagellum, a typical axoneme with a 9 + 9 + 2 microtubular complex, and two mitochondrial derivatives are present in both species. Those of S. optatus have a peculiar shape with the apical side, in cross-section, displaying pointed corners. Two small accessory bodies are located between the axoneme and the two mitochondrial derivatives.

New Findings on the Sperm Structure of Tenebrionoidea (Insecta, Coleoptera).

The sperm ultrastructure of a few representative species of Tenebrionoidea was studied. Two species belong to the Mordellidae (Mordellistena brevicauda and Hoshihananomia sp.), one species to Oedemeridae (Oedemera nobilis), and one species to Tenebrionidae (Accanthopus velikensis). It is confirmed that Mordellidae are characterized by the lowest number of spermatozoa per cyst (up to 64), a number shared with Ripiphoridae. In contrast, in the two other families, up to 512 spermatozoa per cyst are observed, the same number present, for example, in Tenebrionidae. Also, as in the other more derived families of tenebrionoids studied so far, during spermatogenesis in O. nobilis and A. velikensis, sperm nuclei are regularly distributed in two sets at opposite poles of the cysts. On the contrary, the Mordellidae species do not exhibit this peculiar process. However, during spermiogenesis, the bundles of sperm bend to form a loop in their median region, quite evident in the Hoshihananomia sp., characterized by long sperm. This process, which also occurs in Ripiphoridae, probably enables individuals to produce long sperm without an increase in testicular volume. The sperm looping could be a consequence of the asynchronous growth between cyst size and sperm length. The sperm ultrastructure of the Mordellidae species reveals that they can be differentiated from other Tenebrionoidea based on the shape and size of some sperm components, such as the accessory bodies and the mitochondrial derivatives. They also show an uncommon stiff and immotile posterior flagellar region provided with only accessory tubules. These results contribute to a better knowledge of the phylogenetic relationship of the basal families of the large group of Tenebrionoidea.

The sperm ultrastructure of members of basal Tenebrionoidea (Coleoptera).

The sperm ultrastructure of some beetles of Tenebrionoidea was studied with particular attention to those of the Ripiphoridae, Mordellidae, and Meloidae. These three groups are often thought to form a clade, which is the sister group of the remaining Tenebrionoidea. The testes of the two former families have thinner but longer spermatic cysts containing fewer and longer sperm. Within each cyst all sperm cells have the same orientation, but cross sections showed that the orientation of the axonemes alternate between adjacent cysts, possibly due to the cysts bending on themselves. In both families the sperm has a bilayered acrosome and the flagellum, which shows mitochondrial derivatives starting laterally to the nuclear base, has a typical 9 + 9+2 axoneme with accessory tubules provided with 16 protofilaments in their wall, and well-structured triangular shaped accessory bodies. In Mordellistena sp (Mordellidae) sperm, both mitochondrial derivatives and accessory bodies are somewhat asymmetrical. Moreover, the flagellum shows a very thin and long tail end provided with only accessory tubules. Meloidae species have testes with thicker sperm cysts containing numerous shorter sperm. Within the individual cysts the sperm flagella exhibit an alternating orientation of their axonemes as consequence of a peculiar spermatogenetic process. The flagellar structure is similar to that of the above-mentioned species, but the accessory bodies are not well defined and constituted by fuzzy material. In Mylabris hieracii (Meloidae) sperm, the acrosome is flat with a conspicuous perforatorium and its nucleus has a peculiar quadrangular section. Berberomeloe majalis sperm has a large acrosome with an unusual pentagonal perforatorium. The centriolar structure of Mylabris variabilis shows a complex of dense radial links connecting the microtubular structures to the plasma membrane. These results suggest that Ripiphoridae have a closer relationship with Mordellidae than with Meloidae. These findings are in agreement with results obtained with molecular data.

The mitogenome of the true bug Nysius cymoides (Insecta, Heteroptera) and the phylogeny of Lygaeoidea.

The complete mitochondrial genome of the true bug (Homoptera) Nysius cymoides (Spinola, 1837) is herein described and used for phylogenetic comparison with other species of Lygaeoidea. The mtDNA has a gene order and other molecular features typically observed in hexapods, and a long A + T-rich region, due to the occurrence of several repeat units. The phylogenetic analyses support the monophyly of all families except Rhyparochromidae.

The sperm ultrastructure of Pytho depressus (Linnaeus, 1767) (Coleoptera, Pythidae).

The sperm ultrastructure of Pytho depressus (Pythidae) is described in this study. The sperm are short cells, about 85-90 µm long, with an acrosome consisting of three layers, a cylindrical nucleus, which at its base has the initial region of two mitochondrial derivatives. The flagellum has two well-developed triangular accessory bodies, and a 9 + 9+2 axonemal pattern with accessory tubules provided with 16 protofilaments in their wall. The structure and shape of the accessory bodies are diagnostic characters within the superfamily. The sperm morphology of P. depressus can be easily distinguished from those of Ripiphoridae, Meloidae and Tenebrionidae. The P. depressus sperm are organized in cysts as in other species of the group but the sperm are not well aligned and show an antiparallel orientation, a feature also observed in other tenebrionids. The phylogenetic implications of the observed sperm features are discussed in the context of comparative sperm ultrastructure of other insect species.

EZmito: a simple and fast tool for multiple mitogenome analyses.

Complete mitochondrial genome data are frequently applied to address phylogenetic/phylogeographic issues at different taxonomic levels in ecology and evolution. While sample preparation/sequencing is becoming more and more straightforward thanks to dropping costs for next-generation sequencing (NGS), data preparation and visualization remains a manually intensive step that may lead to errors if improperly conducted. We have elaborated, and here introduce, EZmito, a simple and intuitive, freely accessible Web Server aimed at automating some of these tasks. EZmito is divided into three main tools: EZpipe that assembles DNA matrices for phylo-mitogenomic analyses; EZskew that calculates genome, strand, and codon nucleotide compositional skews and EZcodon which computes Relative Synonymous Codon Usage statistics as well as amino acid usage frequency over multiple mitogenomes. Output is produced in tabular format as well as publication-quality graphics.

The ultrastructure of sperm and female sperm storage organs in the water strider Gerris lacustris L. (Heteroptera) and a possible example of genital coevolution.

The fine structural organization of the male and the female inner reproductive apparatuses of the water-strider Gerris lacustris was studied. The sperm of the species shows a long helicoidal acrosome provided with longitudinal tubules, and a short nucleus. The flagellum is characterized by crescent mitochondrial derivatives and a 9 + 9 + 2 axoneme, as occurs in all Heteroptera. The female reproductive apparatus is characterized by an extremely long spermathecal duct, filled with sperm, which plays the role of the main sperm storage organ. The duct has a thin epithelium surrounded by a complex of secretory and duct-forming cells. The spermathecal duct flows into the gynatrial sac. This region, together with the fertilization chamber, exhibits a simple epithelium with deep apical plasma membrane invaginations, and it does not show conspicuous secretions. The basal cell region shows plasma membrane infoldings forming thin cytoplasmic bands hosting mitochondria and large intercellular spaces. This organization is typical of epithelia active in fluid reabsorption. Two lateral large gynatrial glands open into the gynatrial sac. Such glands also exhibit secretory and duct forming cells. The same structure of these glands is also present along the proximal region of the fecundation canal. The duct forming cells of these regions have very wide ducts with peculiar cuticular finger-like structures at their opening into the gland duct lumen. The results of the present study suggest the occurrence of a coevolution between the sperm and the spermathecal duct lengths.

Coevolution between female seminal receptacle and sperm morphology in the semiaquatic measurer bug Hydrometra stagnorum L. (Heteroptera, Hydrometridae).

The coevolution between sperm length and size of the female sperm-storage organs is described for the first time within Heteroptera. The long sperm of the measurer bug Hydrometra stagnorum is characterized by the unusually long acrosome with its anterior region helically arranged, and by a very short nucleus. The sperm flagellum has a 9 + 9+2 conventional axoneme and crystallized mitochondrial derivatives. The female spermatheca consists of an extraordinarily long spermathecal duct ending with an apical spermathecal bulb into which flows also the secretions of a relatively short spermathecal gland. Both spermathecal duct and gland have a thin epithelium lined by a cuticle, beneath which a complex of secretory and duct forming cells are present. The secretions of these two structures flow into the apical spermathecal bulb. A thick layer of muscle fibers surrounds the epithelium. These results confirm the opinion that the dimensions of the female reproductive sperm-storage organs are able to drive the sperm morphology.

Ecosystem Birth Near Melting Glaciers: A Review on the Pioneer Role of Ground-Dwelling Arthropods.

As glaciers retreat, their forelands represent "natural laboratories" for the study of primary succession. This review describes how certain arthropods conquer pristine ground and develop food webs before the establishment of vascular plants. Based on soil samples, pitfall traps, fallout and sticky traps, gut content studies, and some unpublished data, we compare early arthropod succession on glacial forelands of northern Europe (Iceland, Norway including Svalbard, and Sweden) and of the Alps (Austria, Italy). While macroarthropod predators like ground beetles (Coleoptera: Carabidae), harvestmen (Arachnida: Opiliones), and spiders (Arachnida: Araneae) have usually been considered as pioneers, assumed to feed on airborne prey, this review explains a different pattern. Here, we highlight that springtails (Collembola), probably feeding on biofilm made up of algae or cyanobacteria, are super-pioneers, even at high altitudes and under arctic conditions. We also point out that macroarthropod predators can use locally available prey, such as springtails or non-biting midges (Diptera: Chironomidae). Pioneer arthropod communities vary under different biogeographical and climatic conditions. Two pioneer food webs, from northern Europe and the Alps, respectively, differed in structure and function. However, certain genera and orders were common to both. Generalists and specialists live together in a pioneer community. Cold-adapted specialists are threatened by glacier melting.

Evidence for Cryptic Diversity in the "Pan-Antarctic" Springtail Friesea antarctica and the Description of Two New Species.

The invertebrate terrestrial fauna of Antarctica is being investigated with increasing interest to discover how life interacts with the extreme polar environment and how millions of years of evolution have shaped their biodiversity. Classical taxonomic approaches, complemented by molecular tools, are improving our understanding of the systematic relationships of some species, changing the nomenclature of taxa and challenging the taxonomic status of others. The springtail Friesea grisea has previously been described as the only species with a "pan-Antarctic" distribution. However, recent genetic comparisons have pointed to another scenario. The latest morphological study has confined F. grisea to the sub-Antarctic island of South Georgia, from which it was originally described, and resurrected F. antarctica as a congeneric species occurring on the continental mainland. Molecular data demonstrate that populations of this taxon, ostensibly occurring across Maritime and Continental Antarctica, as well as on some offshore islands, are evolutionarily isolated and divergent and cannot be included within a single species. The present study, combining morphological with molecular data, attempts to validate this hypothesis and challenges the taxonomic status of F. antarctica, suggesting that two additional new species, described here as Friesea gretae sp. nov. and Friesea propria sp. nov., are present in Continental Antarctica.

New findings on the sperm ultrastructure of Carabidae (Insecta, Coleoptera).

The sperm structure of several species belonging to different tribes of the large Carabidae family is described. Some species of Nebriinae, such as Nebria brevicollis and Notiophilus biguttatus, have free conventional insect sperm. Their sperm type can be regarded as the ancestral model for Carabidae. All the other species examined, either with isolated sperm such as Calomera nemoralis, Scarites sp., Duvalius andreinii and Anillus florentinus or with spermatozeugmata and sperm associated to spermatostyles such as Typhloreicheia usslaubi, Brachinus italicus, Carabus convexus, Calathus fuscipes, Calathus montivagus, and Paraphorus mendax, showed sperm with long nucleus and a parallel axoneme running the length of the tail starting from the apical bell-like acrosome. C. nemoralis, like Cicindela campestris previously studied, has a sperm structure similar to that of several other Carabidae, confirming their correct assignment to the family. C. convexus has the same sperm structure as previously studied C. preslii and C. interstitialis, indicating that the spermatozeugmata of the group consist only of an apical cap in which the anterior sperm regions are embedded. Unlike other Carabidae with spermatozeugmata, Carabini have the typical sperm organization with acrosome, nucleus and flagellum in a regular sequence. A. florentinus, (Trechinae) shows major differences, such as the absence of an acrosome and an extremely long nucleus that reaches the end of the tail in close association with the axoneme. T. usslaubi (Scaritinae) has slender spermatozeugmata with orderly quartets of sperm. The posterior region of the sperm tail is also unusual, showing a perfect circular section and a plasma membrane reinforced by a dense underlying layer. The present observations confirm that spermatozeugmata, can vary in shape and size among different species of the Carabidae. Such diversity may be the result of the male reproductive strategy, different in each species, that enhances the efficiency of sperm transfer to the female.

The complete mitochondrial genome of the springtail Allacma fusca, the internal phylogenetic relationships and gene order of Symphypleona.

Symphypleona (sensu stricto) are a group of Collembola (=springtails) that, despite displaying some variation in gene order, have been poorly investigated under the phylomitogenomic perspective. How families and subfamilies of this taxon are evolutionary related is still partially unknown. For this reason we sequenced, and herein described, the complete mitochondrial genome sequence of Allacma fusca (Sminthuridae). This sequence, alongside others from the literature, is here used to study the phylogenetic relationships among Symphypleona.

The mitogenome of the jumping bristletail Trigoniophthalmus alternatus (Insecta, Microcoryphia) and the phylogeny of insect early-divergent lineages.

The complete mitochondrial genome of the machilid Trigoniophthalmus alternatus (Silvestri 1904) is herein described and applied to phylogenetic analyses, inclusive of the most early-divergent lineages of hexapods. Both gene content and order generally conform with the organization of the arthropods' mitochondrial genome. One gene translocation involving trnA is the autapomorphic character observed in this species. Another peculiar molecular feature is the long size of the A + T-rich region, due to the occurrence of repeat units. The phylogenetic analyses support the typical placement, along the hexapods' tree, of Ectognatha, Monocondylia and Dicondylia, with Diplura as the adelphotaxon of all true insects.

The mitochondrial genome of the springtail Bourletiella arvalis (Symphypleona, Collembola).

The complete mitochondrial genome of the springtail Bourletiella arvalis (Fitch, 1863) is herein described and applied to a Bayesian phylogenetic analysis, inclusive of all the Collembola mitochondrial DNAs sequenced so far. The gene content and order, as well as the nucleotide composition, conform with the well-known features of hexapods' mitochondrial genomes. The phylogenetic analysis supports the monophyly of Collembola, Poduromorpha, Entomobryomorpha and Symphypleona. However, no mtDNA from Neelipleona is available to date, therefore limiting the application of mitochondrial genomes to further investigate springtail systematics.

Redescription and neotype designation of the Antarctic springtail Folsomotoma octooculata (Collembola: Isotomidae).

The growing interest in Antarctic collembolan species has led us to re-evaluate both the morphological and molecular aspects of different species adapted to the extreme Antarctic environment. The genus Folsomotoma includes nine species mainly distributed in Antarctic and subantarctic areas and a few additionally from Australia and South America. We herein designate the neotype of Folsomotoma octooculata (Willem 1901) and redescribe its morphological characters with particular emphasis on the sensory and ordinary chaetotaxy. Furthermore, based on analyses of both our new and previously published morphological data for F. octooculata, we propose a systematic key of the species for the genus.

A new cave-dwelling species of Deuteraphorura from northern Italy (Collembola, Onychiuridae).

A new cave-dwelling species, Deuteraphorura pierolucciisp. n., is described from northern Italy. The size of adult specimens, number, and distribution of dorsal and ventral pseudocelli, and number of compound vesicles of the post antennal organ were used to distinguish it among other congeneric species.

Spermiogenesis and sperm ultrastructure of Machilontus sp (Insecta: Archaeognatha) with phylogenetic consideration.

The sperm structure of the jumping bristletail Machilontus sp has been described. The species shares several sperm characteristics with other genera of the same order Archaeognatha. During late spermiogenesis the spermatid bends at half of its length with the two limbs closely apposed within the same plasma membrane. The sperm has a helicoidal bi-layered acrosome with a filamentous perforatorium and a long nucleus. The elongated flagellum consists of an axoneme with 9 accessory microtubules external to the 9+2, two rows of conventional mitochondria and two accessory bodies. The accessory bodies are located lateral to the axoneme and are probably responsible for the shifting of the accessory tubules in two opposite groups of 5 and 4 tubules, respectively. These sperm characteristics represent common traits of all Archaeognatha.