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Plastic pollution has become a serious issue of global concern, and biodegradation of plastic wastes is representing one attractive environment-friendly alternative to traditional disposal paths. It is known that insects are involved in the plastic polymer degradation process, with reported evidence of tenebrionid beetle larvae capable to degrade polystyrene (PS), one of the most used plastics worldwide. Recently, a ribosomal RNA based survey on the insect gut microbiota of the lesser mealworm Alphitobius diaperinus has revealed differentially abundant microbial taxa between PS-fed larvae and control group. Following these findings, an enrichment bacterial culture was set up in liquid carbon-free basal medium with PS film as sole carbon source using PS-fed larvae of A. diaperinus as inoculum. After two-months the culture was analysed both by molecular and culture-based methods. Isolated bacteria which had become prevalent under the selective enrichment conditions resulted ascribable to three taxonomic groups: Klebsiella, Pseudomonas, and Stenothrophomonas. The predominance of these groups in PS-fed larvae was confirmed by using bacterial 16S rRNA gene amplicon sequencing, and it was consistent with the results of previous reports. Isolated bacteria were able to attach to PS surfaces and SEM observations showed the presence of thin fibrillar structures connecting the bacterial cells to the abiotic surface.

We describe in this paper the morphology of the male reproductive system, the spermiogenesis and the sperm ultrastructure of the drugstore beetle S. paniceum (L) (Coleoptera, Ptinidae). The male reproductive apparatus has a pair of long testes with six follicles, filled with cysts, each containing 128 germ cells. The sperm is about 480 μm long and is endowed of an apical spiral cylindrical acrosome, 1.7 μm long. The nucleus, 25 μm long, is also cylindrical at maturity. During spermiogenesis the nuclear envelope gives rise to two ovoid membranous structures flanking the nucleus for its whole length. The centriole adjunct material is configurated as concentric electron-dense rings. The flagellar components have a helical array; the axoneme has the typical 9 + 9+2 pattern for most of its length, but in the posterior flagellar region it modifies greatly its ultrastructure and is embedded in an electron-dense material. The two accessory bodies have a crescent-moon shape in the mature sperm, but at the beginning of spermiogenesis they are small structures lateral to the axoneme, later assuming a crescent-moon structure. A flattened cistern located between the axoneme and the plasma membrane is present in the early spermatids, but soon it fuses with the plasma membrane. Two symmetrical, largely crystallized mitochondrial derivatives are present for most of the flagellar length. S. paniceum and Lasioderma serricorne share several sperm structures. Ultrastructural results and molecular data confirm a close relationship between these two species and support their positioning within Ptinidae.

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.

Relatively few studies have focused on evolutionary losses of sexually selected male traits. We use light and electron microscopy to study the male and female reproductive anatomy of Apotomus ground beetles (Coleoptera, Carabidae), a lineage that we reconstruct as likely having lost sperm conjugation, a putative sexually selected trait. We pay particular attention to the structure of the testes and spermatheca. Both of these organs share a strikingly similar shape-consisting of long blind canals arranged into several concentric overlapping rings measuring approximately 18 mm and 19.5 mm in total length, respectively. The similarity of these structures suggests a positive evolutionary correlation between female and male genital organs. Males are characterized by unifollicular testes with numerous germ cysts, which contain 64 sperm cells each, and we record a novel occurrence of sperm cyst "looping", a spermatogenic innovation previously only known from some fruit fly and Tenebrionid beetle sperm. The sperm are very long (about 2.7 mm) and include an extraordinarily long helicoidal acrosome, a short nucleus, and a long flagellum. These findings confirm the structural peculiarity of sperm, testis, and female reproductive tract (FRT) of Apotomus species relative to other ground beetles, which could possibly be the result of shifts in sexual selection.

The fine structure of the female reproductive organs of the diving beetle Scarodytes halensis has been described, with particular attention to the complex organization of the spermatheca and the spermathecal gland. These organs are fused in a single structure whose epithelium is involved in a quite different activity. The secretory cells of the spermathecal gland have a large extracellular cistern with secretions; duct-forming cells, by their efferent duct, transport the secretions up to the apical cell region where they are discharged into the gland lumen. On the contrary, the spermatheca, filled with sperm, has a quite simple epithelium, apparently not involved in secretory activity. The ultrastructure of the spermatheca is almost identical to that described in a closely related species Stictonectes optatus. Sc. halensis has a long spermathecal duct connecting the bursa copulatrix to the spermatheca–spermathecal gland complex. This duct has a thick outer layer of muscle cells. Through muscle contractions, sperm can be pushed forwarding up to the complex of the two organs. A short fertilization duct allows sperm to reach the common oviduct where eggs will be fertilized. The different organization of the genital systems of Sc. halensis and S. optatus might be related to a different reproductive strategy of the two species.

Large-scale production of the black soldier fly [Hermetia illucens (L.) (Diptera: Stratiomyidae)] for use as aquaculture and poultry feed has developed into a global industry. Successful commercialization of the black soldier fly relies on optimizing the production of fecund adults. However, current mass-rearing protocols result in variable production of fertile eggs. To help lay a foundation for a better understanding of factors that may play a role in this variability, the morphology of the black soldier fly male reproductive tract and spermatozoa, associated spermatogenesis process, impact of age on the process, and the female spermatheca morphology were examined with various microscopic techniques (e.g., scanning electron microscope, transition electron microscope, and dissecting scope). The gross morphology of the male reproductive tract and female spermatheca appear to be similar to those found in other brachyceran flies. Male spermatozoa are long (~860 µm overall, ~8 µm head), apparently motile, and possess flagella with a typical 9 + 9 + 2 axoneme triplets. Germ cells go through incomplete mitotic divisions surrounded by somatic cyst cells in the testes. Spermatogenesis appears to be initiated during immature development (cryptocephalic pupa stage). From

In social wasps, female lifespan depends on caste and colony tasks: workers usually live a few weeks while queens as long as 1 year. Polistes dominula paper wasps infected by the strepsipteran parasite Xenos vesparum avoid all colony tasks, cluster on vegetation where parasite dispersal and mating occur, hibernate and infect the next generation of wasp larvae. Here, we compared the survival rate of infected and uninfected wasp workers. Workers’ survival was significantly affected by parasite sex: two-third of workers parasitized by a X. vesparum female survived and overwintered like future queens did, while all workers infected by a X. vesparum male died during the summer, like uninfected workers that we used as controls. We measured a set of host and parasite traits possibly associated with the observed lifespan extension. Infected overwintering workers had larger fat bodies than infected workers that died in the summer, but they had similar body size and ovary development. Furthermore, we recorded a positive correlation between parasite and host body sizes. We hypothesize that the manipulation of worker’s longevity operated by X. vesparum enhances parasite’s fitness: if workers infected by a female overwinter, they can spread infective parasite larvae in the spring like parasitized gynes do, thus contributing to parasite transmission.

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.

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.

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 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 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.

The sperm of three coccinellid species belonging to the subfamilies Chilocorinae, Coccinellinae and Epilachninae were studied under light and transmission electron microscopy. The basic sperm structure of these ladybirds is common to that of the other previously studied species, especially the acrosome in front of the basal body and not the nucleus, with this latter running parallel with the flagellar components. In the Chilocorinae Platynaspis luteorubra (Platynaspidini) the sperm are of the type 1, as in Scymnini and Coccinellini, since they exhibited a cylindrical basal body with 9 + 0 and then 9 + 2, microtubules continuing further in an initial flagellar portion with the only axoneme devoid of accessory structures. The sperm exhibits a thin nucleus and mitochondrial derivatives. Such uniformity of sperm ultrastructure could be indicative of the occurrence of a close relationship between Platynaspidini and Scymnini as also proposed in the previous studies. Conversely, they differ markedly from the sperm of type 3 observed in the Chilocorini Exochomus quadripustulatus. In the Coccinellini Propylea quatuordecimpunctata the sperm are also of the type 1, but they can be easily differentiated from those of the other Coccinellinae studied so far, because of their very short acrosome without the posterior extension, the relatively thicker mitochondrial derivatives and the cylindrical nucleus. Epilachna clandestina sperm resemble those of E. quadripustulatus but differ from them, because they exhibit an elliptical nucleus which is anteriorly very thin, and the asymmetrical mitochondrial derivatives in the anterior extremity, with the greater one starting at the same level of the basal body, rather than at the nucleus level, as it occurs in E. quadripustulatus. Because of the differences observed in the sperm ultrastructure we propose a new sperm type (Type 4) for E. clandestina. This study on ladybird spermatozoon ultrastructure clearly indicates that the current classifications of coccinellids do not reflect the natural history of this well-known insect family.

The sperm structure and spermiogenesis of the morphologically peculiar heteropteran Coptosoma scutellatum has been examined with electron microscopy. The sperm has a short monolayer acrosome, a cylindrical nucleus, a 9+9+2 axoneme and two mitochondrial derivatives with crystallized matrix. It shows the synapomorphies typical of the group, consisting of (a) two crystallized areas in the mitochondrial derivatives; (b) the presence of two bridges connecting the intertubular material of the flagellar axoneme to the flattened cisterns adhering to the inner sides of the mitochondrial derivatives; (c) the absence of accessory bodies. The most interesting feature is the presence, in the early spermatid, of an extended centriole adjunct material with the shape of a large ribbon-like structure. This material surrounds the posterior nuclear region and extends anteriorly along the nucleus. It is a microtubule-organizing-center (MTOC) producing the many microtubules surrounding the sperm components At the end of spermiogenesis, the centriole adjunct material modifies into two kidney-like structures accompanying the nuclear posterior end and surrounding the modified centriole.

In the present study, we describe the sperm morphology of 11 species of Elateriformia (9 elaterids, 1 lampyrid and 1 buprestid) using transmission electron microscopy. All species exhibited sperm that is not usually observed in insects in general. The most highlighted features are the displacement of the nucleus running parallel to the flagellar components, hitherto observed only in coccinellid and carabid beetles, and the presence of thin and dense structures along the nucleus, probably derived from the centriole adjunct, a feature that is so far exclusive to these insects. The other structures are a typical axoneme for insects with 9 + 9 + 2 microtubules, in a position diametrically opposite relative to the nucleus, two slender, symmetrical mitochondrial derivatives and a pair of discrete accessory bodies. This arrangement provides a bilaterally symmetrical flagellum, which favourably influences sperm hydrodynamics, as will be discussed. The occurrence of this unusual structural arrangement in the sperm of species from superfamilies that are phylogenetically as distant as Elateroidea and Buprestoidea support the monophyly of the infraorder Elateriformia, as proposed by some previous molecular studies.

Popillia japonica is an invasive scarab beetle native to Japan that in 1916 invaded New Jersey in USA. From that moment onwards, the insect has spread invading several US states, Canada, the Azores, Italy and, recently, Switzerland. It is a severe agricultural pest included in the EU priority pest list being able to feed on more than 300 plant species and having an important biotic potential. The general morphology of the reproductive apparatus shows paired testes, each of them having six testicular lobes grouped in threes. From the ventral part of each testicular lobe, each containing about 20 follicles, an efferent vessel originates that fuses with the other efferent vessels to form the deferent duct. A pair of long tubular accessory glands is present. The deferent ducts and accessory glands fuse together into an ejaculatory duct before entering the aedeagus. The sperm is a typical pterygote sperm, 110 μm long, composed of a head and a tail. In the head a three-layered acrosome of about 6 μm in length and a nucleus of about 18 μm long are present. During sperm maturation two C-shaped structures appear in the cytoplasm from the opposite sides of the nucleus that then disappear in late spermatids. In the tail a typical 9 + 9 + 2 flagellar axoneme and two mitochondrial derivatives are present. Moreover, in the head-tail transition region the centriolar adjunct forms a sheath from which three elongated accessory bodies originate. Two of these accessory bodies are placed alongside the axoneme, whilst the third one is placed beneath the mitochondrial derivatives. Mature sperm are grouped in cysts containing about 256 sperm cells. A morphological comparison with related species is provided.

The ultrastructure of the complex organisation of the spermatozoa in Harmonia axyridis and Adalia decempunctata (Coccinellidae) was studied, with particular emphasis on the origin of the anterior shifting of the axonemal structure, which becomes parallel to the nucleus in the sperm flagellum. In studying the spermiogenesis, a centriolar remodelling was observed with the long centriole, present in the early spermatids, transformed in the spermatozoa into an exceptionally long and narrowed basal body (about 0.16 × 3.5–4.0 μm long) displaying a 9 + 0 microtubular pattern in the proximal part and a 9 + 2 pattern in the following part; this is a characteristic not observed in any other pterygotan insect. The sperm also have a very long acrosome surrounded by a dense layer of material extending along the whole basal body. These two uncommon features were discussed in the light of sperm movement.

Sperm structure of seven species from different Carabidae tribes was studied. Carabus preslii and Carabus granulatus interstitialis form sperm bundles (spermatozeugmata) in which the heads of conventional insect sperm cells are embedded in the apical cap (spermatostyle), leaving the posterior flagella free. On the contrary, Pterostichus morio, Pterostichus melas, Pterostichus melanarius and Amara aulica form complex spermatozeugmata bearing conspicuous axial spermatostyles associated with lateral groups of sperm. Individual sperm cells are contained in chambers delimited by laminar extensions of the cortical region of the spermatostyle. Demetrias atricapillus shares the general spermatozeugma structure with the above species, but the anterior spermatostyle region has a different structure and posteriorly, the lateral groups of sperm are separated from the spermatostyle but remain connected to it by peduncles consisting of extensions of the cortical region of the spermatostyle. The sperm of the species examined in the study consist of a short, flat acrosomal cap and, with the exception of the Carabus species, show long nuclei which extend parallel to the axonemes, along the flagella. Two relatively small mitochondrial derivatives and two small accessory bodies flank the axoneme. These bodies become very thick in the posterior flagellar region of D. atricapillus. The study revealed different spermatozeugma models and different sperm organization in the Carabidae family.

Previous studies on the spermatogenesis of tenebrionid beetles showed the unusual formation of two antiparallel sperm bundles per cyst. In this work we reported this feature also in Tribolium castaneum using light and transmission electron microscopy. The sperm structure of T. castaneum, similar to other tenebrionids, consists of a three-layered acrosome, an elongated nucleus and a flagellum with a 9 + 9 + 2 axoneme, two accessory bodies and two asymmetric mitochondrial derivatives. The presence of two antiparallel sperm bundles per cyst also in Meloidae and Rhipiphoridae suggests that it is a strong trait synapomorphic for Tenebrionoidea. The huge degeneration of whole sperm cells in several cysts of testes during spermiogenesis is also described.

The sperm structure of several ladybird species belonging to different subfamilies of Coccinellidae was studied. Three main sperm types were clearly recognized, and were characterized by differences in acrosomal length, the presence of a dense coat around the acrosome, the length of the basal body, the amount of the centriole adjunct material, and the diameter of the mitochondrial derivatives. However, the whole group shares a pattern of the posterior sperm region uncommon for insects, in which the axoneme and other flagellar components are running parallel with the nucleus. As a general conclusion, this study has revealed an inconsistency between the sperm structure and the systematics of the group, indicating that the generic concepts within the group do not reflect a natural classification, a statement also shared by molecular studies.

We studied spermatogenesis and spermiogenesis in Adalia decempunctata (L), a beetle of the Coccinellidae family. The spermatocyte exhibits two centrioles which elongate to form a pair of primary cilia. A novel structure, appearing in cross sections as a dense droplet, is observed near the long centriole during spermiogenesis, and is soon accompanied by a procentriole (PCL). PCL structure consists of singlet microtubules, a central tubule and an incomplete cartwheel. The PCL persists until the end of spermiogenesis, when it vanishes together with the dense droplet. The sperm has an exceptionally long basal body and the nucleus is disposed parallel to the flagellar components, a peculiar trait shared by other species of the coccinellid group. The presence of a procentriole suggested by the use of antibodies is discussed.

The springtail Collembola are characterized by having rolled spermatozoa, with a long cylindrical extracellular structure adhering to the acrosome. This structure is produced by the secretory activity of the testes epithelial cells at almost the end of spermiogenesis. At the beginning of its formation, it is a thin extension with a helical wall and a dense axial region. Later the cylindrical structure shows an inner organization which is different in the several species examined: species of Entomobryidae contain material with a paracrystalline structure, whilst some of Symphypleona contain ovoid structures. The outer envelope of the extracellular structure consists of two overlapped layers orthogonally arranged, clearly identified by cryo-preparations. Immunoblot analysis and lectin stainings have indicated that the cylindrical structure has a glycoproteic composition. As the structure is no longer visible after the sperm transfer into the female spermatheca, it is suggested that it could contain enzymes able to activate the sperm unwinding process and possibly allowing the reacquisition of sperm motility.

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.

Sperm of the dictyopteran key taxon Cryptocercus punctulatus was examined. It has largely maintained a blattodean groundplan condition, with a three-layered acrosome, an elongate nucleus, a single centriole, a conspicuous centriole adjunct material, two connecting bands (=accessory bodies), and a long functional flagellum with a 9+9+2 axoneme provided with accessory tubules with 16 protofilaments and intertubular material. These sperm characters are shared with several other polyneopterans. The sperm of C. punctulatus is very similar to what is found in Periplaneta americana and species of other groups of roaches, including the sperm of Loboptera decipiens described here for the first time. The general sperm organization here described can be assumed for the groundplan of Insecta and Pterygota. The following evolutionary path can be suggested: after the split between Cryptocercidae and the common ancestor of Isoptera, the typical pattern of sperm formation was altered very distinctly, resulting in a duplication or multiplication (Mastotermitidae) of the centrioles. Mastotermes has maintained a certain sperm motility, but with a very unusual apparatus of multiple flagella with a 9+0 axoneme pattern. After the split into Mastotermitidae and the remaining Isoptera, sperm motility was completely abandoned, and different modifications of sperm components occurred, and even the loss of the sperm flagellum. J. Morphol. 276:361–369, 2015. © 2014 Wiley Periodicals, Inc.

The Ephemeroptera sperm axoneme is devoid of outer dynein arms (ODA) and exhibits a pronounced modification of the central pair complex (CPC), which is substituted by the central sheath (CS): a tubular element of unknown molecular composition. We performed a detailed ultrastructural analysis of sperm axonemes in the genera Cloeon and Ecdyonurus using quick-freeze, deep-etch electron microscopy, showing that the loss of the conventional CPC is not only concomitant with the loss of ODA, but also with a substantial modification in the longitudinal distribution of both radial spokes (RS) and inner dynein arms (IDA). Such structures are no longer distributed following the alternation of different repeats as in the 9 + 2 axoneme, but instead share a 32 nm longitudinal repeat: a multiple of the 8 nm repeat observed along the CS wall. Differently from the conventional CPC, the CS and the surrounding RS possess a ninefold symmetry, coherently with the three-dimensional pattern of motility observed in Cloeon free spermatozoa. Biochemical analyses revealed that ultrastructural modifications are concomitant with a reduced complexity of the IDA heavy chain complement. We propose that these structural and molecular modifications might be related to the relief from the evolutionary constraints imposed by the CPC on the basal 9 + 9 + 2 axoneme and could also represent the minimal set compatible with flagellar beating and progressive motility mechanically regulated as suggested by the geometric clutch hypothesis. © 2014 Wiley Periodicals, Inc.

The external gestation of sexually ripe females of the species Exogone naidina (Polychaeta, Syllidae) is described by means of SEM and TEM analysis. The eggs, embryos and juveniles are attached in close vicinity of each parapodial complex in a position immediately below each ventral cirrus and are connected to the female by a cup like structure. The formation of this adhesive disk is linked to secretory cells scattered between dermal cells of ripe female. This adhesive disk is present only in sexually mature animals and is considered as epitokous structure. The evolutive significance of ventral and dorsal attachment found within the Exogoninae is also discussed. © 2003 Elsevier Ltd. All rights reserved.

The spermiogenesis, the sperm structure and the sperm motility of Marchalina hellenica (Gennadius) were examined. In the early spermiogenesis a centriolar apparatus was identified, but this structure is not involved in the production of the sperm flagellum. As in other Coccoidea, the flagellar axoneme originates by the activity of the thickened tip of the numerous microtubules surrounding the nuclear anterior region close to the periphery of the cell. This region pushes against a narrow cytoplasmic layer, giving rise to a papilla. In this region a novel structure, consisting of a regular network of thin filaments, arranged orthogonally to the bundle of microtubules, is visible. The sperm flagellum consists of a series of about 260 microtubules, regularly arranged in rings around the axial nucleus. This latter extends in the middle part of the sperm length. As usual in scale insects, sperm form a bundle, which in M. hellenica is composed of 64 sperm cells, surrounded by somatic cyst cells. The sperm bundle has an helicoidal array, with a cap of dense material at its apex, lending the anterior and the posterior region of the sperm bundle with a different structural organization. This difference is responsible of the different speed gradient observed in the helical wave propagating along the sperm bundle.

The salivary glands of two species of Zoraptera, Zorotypus caudelli and Zorotypus hubbardi, were examined and documented mainly using transmission electron microscopy (TEM). The results obtained for males and females of the two species are compared and functional aspects related to ultrastructural features are discussed. The salivary glands are divided into two regions: the secretory cell region and the long efferent duct, the latter with its distal end opening in the salivarium below the hypopharyngeal base. The secretory region consists of a complex of secretory cells provided with microvillated cavities connected by short ectodermal ducts to large ones, which are connected with the long efferent duct. The secretory cell cytoplasm contains a large system of rough endoplasmic reticulum and Golgi apparatus producing numerous dense secretions. The cells of the efferent duct, characterized by reduced cytoplasm and the presence of long membrane infoldings associated with mitochondria, are possibly involved in fluid uptaking from the duct lumen.

The rectal pads of a species of the controversial polyneopteran order Zoraptera were examined using histological sections and TEM micrographs. Six pads are present along the thin rectal epithelium. Each pad consists of a few large principal cells surrounded by flattened junctional cells, which extend also beneath the principal cells. The cells are lined by a thin apical cuticle. No basal cells and no cavity have been observed beneath the pad. Principal cells have a regular layer of apical microvilli and are joined by intercellular septate junctions, which are interrupted by short dilatations of the intercellular space. At these levels the two adjacent plasma membranes are joined by short zonulae adhaerentes. In the cytoplasm, a rich system of strict associations between lateral plasma membranes and mitochondria forms scalariform junctions. Rectal pads share ultrastructural features with similar excretory organs of several neopteran groups, in particular with Blattodea (roaches and termites) and Thysanoptera, and are involved in fluid reabsorption and ion regulation.

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.

The structural features of eggs of Zorotypus caudelli Karny are described in detail. The egg is elliptic with long and short diameters of 0.6 and 0.3 mm respectively, and creamy white. The egg shows a honeycomb pattern on its surface, without any specialized structures for hatching such as an operculum or a hatching line. The fringe formed by a fibrillar substance secreted after the completion of the chorion encircles the lateral surface. The egg layer is composed of an exochorion, an endochorion, and a vitelline envelope. The exochorion and endochorion are electron-dense and homogeneous in structure. The exochorion shows a perforation of numerous branching aeropyles. The exo- and endochorion are connected by numerous small columnar structures derived from the latter. The vitelline envelope is very thin and more electron-dense than the chorion. A pair of micropyles is present at the equator on the dorsal side of the egg. Originating at the micropyle, the micropylar canal runs through the chorion obliquely. The structural features of the eggs of Zoraptera were compared with those of other polyneopteran and paraneopteran orders.

Endosymbionts of the Cardinium-like genus are described in the testes and other tissues of the proturan Acerella muscorum (Ionescu). Few endosymbionts are present in the large apical cells of functional testes, but they become numerous at the end of the reproductive cycle. They are also found within sperm cells where induce their degeneration. The Gram-negative endosymbionts are characterized by the presence of microtubule-like structures (MLC) in their cytoplasm. It is suggested a possible role of the endosymbionts in the elimination of degenerating sperm cells when the testes activity is ended, thus somewhat playing a role in the timing of the reproductive cycle of the proturan species.

Sperm ultrastructure and spermiogenesis of the dipluran Japygidae (Japyx solifugus, Metajapyx braueri and Occasjapyx japonicus) and Campodeidae (Campodea sp.) were studied with the aim of looking for potential characters for the reconstruction of the phylogenetic relationships of basal hexapods. Both Japygidae and Campodeidae share a common sperm axonemal model 9+9+2, provided with nine accessory microtubules. These microtubules, however, after their formation lose the usual position around the 9+2 and migrate between the two mitochondria. In Japygidae, four of these microtubules are very short and were observed beneath the nucleus after negative staining and serial sections. Accessory microtubules have 13 protofilaments in their tubular wall. Diplura have a sperm morphology which is very different from that of the remaining Entognatha (Protura+Collembola). On the basis of the present results, the presence of accessory microtubules suggests that Diplura are the sister-group of the Insecta s.s.. Moreover, Japygidae and Campodeidae differ with regards to the relative position of the sperm components, the former having the axoneme starting from beneath the nucleus (above which sits the short acrosome), while the latter having a long apical acrosome and a nucleus running parallel with the proximal part of the axoneme. The present study also allowed to redescribe the male genital system of Japyx.

The general organization of the male genital system, the spermatogenesis and the sperm structure of the proturan Acerella muscorum have been described. At the apex of testis apical huge cells are present; their cytoplasm contains a conventional centriole, a large amount of dense material and several less electron-dense masses surrounded by mitochondria. Spermatocytes have normal centrioles and are interconnected by cytoplasmic bridges. Such bridges seem to be absent between spermatid cells and justify the lack of synchronization of cell maturation. Spermatids are almost globular cells with a spheroidal nucleus and a large mass of dense material corresponding to the centriole adjunct. Within this mass a centriole is preserved. Mitochondria of normal structure are located between the nucleus and the plasma membrane. The spermatids are surrounded by a thick membrane. No flagellar structure is formed. Sperm have a compact spheroidal nucleus, a large cap of centriole adjunct material within which a centriole is still visible. A layer of mitochondria is located over the nucleus. The cytoplasm is reduced in comparison to spermatids; many dense bodies are interspersed with sperm in the testicular lumen. The sperm are small, immotile cells of about 2.5-3microm in diameter.

The sperm tail of bug insects has 9 + 9 + 2 flagellar axonemes and two mitochondrial derivatives showing two to three crystalline inclusions in their matrix. During spermiogenesis, the axoneme is surrounded by a membrane cistern which, at sperm maturity, reduces to two short cisterns on the opposite sides of the axoneme adhering to the mitochondrial derivatives. Filamentous bridges connect the intertubular material of the axoneme to these cisterns. Such bridges, which represent a peculiar feature of bug insects, are resistant to detergent treatment, whereas part of the intertubular material and the inner content of microtubular doublets are affected by the treatment. After freeze-fracture replicas, at the insertion of the bridges to the cisternal membrane, the P-face of this membrane shows a characteristic ribbon consisting of four rows of 11 ± 1 nm staggered intramembrane particles, 13 ± 2 nm apart along each row. The bridges could be able to maintain the axoneme in the proper position during flagellar beating avoiding distortion affecting sperm motility. J. Morphol. 2009. © 2009 Wiley-Liss, Inc.

The "zipper line" of Drosophila melanogaster and of Drosophila species characterized by giant spermatozoa (D. hydei, D. kanekoi and D. bifurca) was studied by electron microscopy using conventional thin-sections, lectin labeling and freeze-fracture replicas. In cross sections the membrane specializations are located either at the level of the short cistern close to the large mitochondrial derivative where a small tuft of glycocalyx is visible or, in species characterized by long spermatozoa, along a cistern beneath the plasma membrane. In correspondence of such cistern, the plasma membrane exhibits a thick and extended glycocalyx. At this level, as well as at the short tuft of D. melanogaster, alpha-mannose residues were detected. The "zipper" of D. melanogaster consists of rows of intramembrane particles longitudinally disposed along the sperm tail and associated with the external face of the plasma membrane. On the protoplasmatic face a narrow ribbon of transversal grooves is visible. Freeze-fracture replicas have revealed, in the region characterized by extended glycocalyx, the presence of a large ribbon of intramembrane particles disposed in parallel transversal rows, associated with the protoplasmatic membrane face. On the complementary external face a ribbon of parallel transversal grooves was observed. It is suggested that membrane specializations are mechanical devices to protect spermatozoa from torsion and bending in the seminal vesicles and then in the female storage organ.

The sperm ultrastructure of a few species of Limoniidae (Limonia nigropunctata; L. nubeculosa; Chionea n. sp.; C. alpina; C. lutescens) was studied. The two species of Limonia have a monolayered acrosome with crystallized material, a three-lobed nucleus in cross section, a ring of centriole adjunct material and a flagellum which consists of a 9 + 9 + 1 axoneme and a single mitochondrial derivative. The central axonemal tubule is provided with 15 protofilaments in its tubular wall, while the accessory tubules have 13 protofilaments and are flanked by the electron-dense intertubular material. The three species of Chionea share a monolayered acrosome, a nucleus with two longitudinal grooves, a centriole adjunct material which surrounds the centriole and the initial part of the axoneme. The axoneme is of conventional type, with 9 + 9 + 2 microtubular pattern, with accessory tubules provided with 13 protofilaments and intertubular material. However, in C. lutescens the accessory tubules start with 15 protofilaments and transform into a tubule with 13 protofilaments. These data are discussed in the light of the phylogenetic relationship between Limoniidae and Tipulidae. For this purpose, the sperm ultrastructure of Nephrotoma appendiculata was also considered comparatively.

Spermatozoa provide an unusual character system, with a limited number of components organized in a single cell. Similar spermatozoa occur in groups widely separated in the phylogenetic tree of Metazoa. Nevertheless, the character system contains phylogenetic information. Hexapoda have acquired spermatophores along with the switch from aquatic to terrestrial habitats, and related to this, a multitude of different sperm types. The aim of this study is a formal evaluation of the phylogenetic information content of spermatozoa. For the first time, sperm characters are coded for formal phylogenetic analyses. Different approaches are used and compared. Mainly due to a high level of homoplasy, the evaluation of sperm characters alone is insufficient for a reconstruction of the phylogeny of the group. Yet, a reliable reconstruction of the evolution of insect sperm is possible when character transformations are assessed using a phylogeny based on extensive molecular data. Important changes took place in the early evolution of Hexapoda. Sperm characters support some major clades (e.g. Hexapoda, Dicondylia, Polyneoptera, Psocodea), but important steps in the evolution are not reflected by transformations of spermatozoa, notably the rise of Pterygota or Holometabola. Important innovations are the formation of mitochondrial derivatives and the acquisition of accessory microtubules. Some features are conservative, whereas others evolved rapidly (e.g. presence or absence of the acrosome vesicle). Some groups are conservative in their sperm features (e.g. Odonata, Heteroptera), whereas the evolution of spermatozoa was distinctly accelerated in others (e.g. Ephemeroptera). The rate of evolution can change drastically in closely related groups. Profound changes in the morphologically uniform Zoraptera underline that sperm evolution can follow a pattern very different from the general somatic morphology. The mode of character reconstruction preferred here will be useful for the evaluation of specialized morphological character systems and strengthen the concept of evolutionary morphology.

The ancestral eukaryotes presumably had an MTOC (microtubule organizing center) which late gave origin to the centriole and the flagellar axoneme. The centrosome of insect early spermatids is in general composed of two components: a single centriole and a cloud of electron-dense pericentriolar material (PCM). During spermiogenesis, the centriole changes its structure and gives rise to a flagellar axoneme, while the proteins of PCM, gamma tubulin in particular, are involved in the production of microtubules for the elongation and shaping of spermatid components. At the end of spermiogenesis, in many insects, additional material is deposited beneath the nucleus to form the centriole adjunct (ca). This material can also extend along the flagellum in two accessory bodies (ab) flanking the axoneme. Among Homoptera Sternorrhyncha, a progressive modification of their sperm flagella until complete disappearance has been verified. In the Archaeococcidae Matsucoccus feytaudi, however, a motile sperm flagellum-like structure is formed by an MTOC activity. This finding gives support to the hypothesis that an evolutionary reversal has occurred in the group and that the cell, when a non-functional centriole is present, activates an ancestral structure, an MTOC, to form a polarized motile bundle of microtubules restoring sperm motility. The presence and extension of the centriole adjunct in the different insect orders is also enlisted.

Thrips spermiogenesis is characterized by unusual features in the differentiating spermatid cells. Three centrioles from which three individual short flagella are initially assembled, make the early spermatid a tri-flagellated cell. Successively, during spermatid maturation, the three basal bodies maintain a position close to the most anterior end of the elongating nucleus, so that the three axonemes are progressively incorporated in the spermatid cytoplasm, where they run in parallel to the main nuclear axis. Finally, the three axonemes amalgamate to form a microtubular bundle. The process starts with the formation of rifts at three specific points in each axonemal circumference, corresponding to sites 1,3,7 and leads to the formation of 9 microtubular rows of different length, i.e. 3 ""dyads"", 3 ""triads"" and 3 ""tetrads"". In the spermatozoon, the nucleus, the mitochondrion and the bundle of microtubules are arranged in a helicoidal pattern. The elongation of the spermatozoon is allowed by the deep anchorage of the spermatid to the cyst cell through a dense mass of material which, at the end of spermiogenesis, becomes a long anterior cylindrical structure. This bizarre ""axoneme"" does not show any trace of progressive movement but it is able to beat. According to the presence of dynein arms, sliding can take place only within each row and not between the rows. The possible molecular basis underlying the peculiar instability of thrips axonemes is discussed in light of the present knowledge on the organization of the axoneme in mutant organisms carrying alterations of the tubulin molecule. Cell Motil.

The sperm of the heteropteran bug Raphigaster nebulosa (Poda) are of two types, differing in length and size of their flagella. The thicker sperm are shorter than the thinner ones and have large mitochondrial derivatives. The presence of virus particles associated with the plasma membrane of thinner sperm is described for the first time; thicker sperm are immune to virus infection. The fact that virus particles are present on thinner sperm only initiates considerations on the transmission of virus.

We present here for the first time a 3D reconstruction of in situ axonemal outer dynein arms. This reconstruction has been obtained by electron tomography applied to a series of tilted images collected from metal replicas of rapidly frozen, cryofractured, and metal-replicated sperm axonemes of the cecidomid dipteran Monarthropalpus flavus. This peculiar axonemal model consists of several microtubular laminae that proved to be particularly suitable for this type of analysis. These laminae are sufficiently planar to allow the visualization of many dynein molecules within the same fracture face, allowing us to recover a significant number of equivalent objects and to improve the signal-to-noise ratio of the reconstruction by applying advanced averaging protocols. The 3D model we obtained showed the following interesting structural features: First, each dynein arm has two head domains that are almost parallel and are obliquely oriented with respect to the longitudinal axis of microtubules. The two heads are therefore positioned at different distances from the surface of the A-tubule. Second, each head domain consists of a series of globular subdomains that are positioned on the same plane. Third, a stalk domain originates as a conical region from the proximal head and ends with a small globular domain that contacts the B-tubule. Fourth, the stem region comprises several globular subdomains and presents two distinct points of anchorage to the surface of the A-tubule. Finally, and most importantly, contrary to what has been observed in isolated dynein molecules adsorbed to flat surfaces, the stalk and the stem domains are not in the same plane as the head. Cell Motil. Cytoskeleton 62:69–83, 2005. © 2005 Wiley-Liss, Inc.

The controversial mating of the strepsipteran Xenos vesparum was studied to investigate the possible sperm routes for fertilization. The female, which is a neotenic permanent endoparasite of Polistes wasps, extrudes only its anterior region, the “cephalothorax,” from the host abdomen. This region has an opening where both mating and larval escape occur. Observations with scanning and transmission electron microscopy revealed spermatozoa not only in the hemocoel, but also in the “ventral canal” (an extragenital duct peculiar to strepsipteran females) and in the “genital ducts” (ectodermal invaginations connecting the ventral canal to the hemocoel) of recently mated females. Xenos vesparum spermatozoa can reach the oocytes either through the hemocoel as a result of a hypodermic insemination, or by moving along the extragenital ducts, which are later used by first instar larvae to escape. The hypothesis of hypodermic insemination is reconsidered in the light of behavioral and ultrastructural evidence. J. Morphol. © 2005 Wiley-Liss, Inc.

Nassonow's gland consists of a number of cells with ducts that open on to the ventral surface of the brood canal in the cephalothoracic region of a neotenic female strepsipteran. The structural organization of the gland is reminiscent of the class 3 of the epidermal gland cells as defined by Noirot and Quennedey [Ann. Rev. Entomol. 19 (1974) 61], which consists of secretory and duct forming cells. The ultrastructure of the Nassonow's gland is described in female Xenos vesparum (Rossi) parasitic in the social wasp Polistes dominulus Christ. The large secretory cells are clustered in groups of three to four, rich in smooth endoplasmic reticulum and produce a secretion made up of lipids. In young females, just before mating, the ultrastructure of the cells and their inclusions indicate that they are active. In old-mated females the Nassonow's gland degenerates. Microvilli line an extracellular cavity and there are pores present in the irregularly thick cuticle of the efferent duct. The small duct forming cells, intermingle with epidermal cells, overlap secretory cells and produce a long efferent duct, the cuticle of which becomes thick close to its opening in the brood canal. Nassonow's gland could be the source of a sex pheromone, which might be capable of attracting the free-living male to a permanently endoparasitic female.

The axonemal organization expressed in the sperm flagella of the cecidomyiid dipteran Asphondylia ruebsaameni is unconventional, being characterized by the presence of an exceedingly high number of microtubular doublets and by the absence of both the inner dynein arms and the central pair/radial spoke complex. Consequently, its motility, both in vivo and in vitro, is also peculiar. Using monoclonal antibodies directed against posttranslational modifications, we have analyzed the presence and distribution of glutamylated and glycylated tubulin isoforms in this aberrant axonemal structure, and compared them with those of a reference insect species (Apis mellifera), endowed with a conventional axoneme. Our results have shown that the unorthodox structure and motility of the Asphondylia axoneme are concomitant with: (1) a very low glutamylation extent in the α-tubulin subunit, (2) a high level of glutamylation in the β-subunit, (3) an extremely low total extent of glycylation, with regard to both monoglycylated and polyglycylated sites, either in α- or in β-tubulin, (4) the presence of a strong labeling of glutamylated tubulin isoforms at the proximal end of the axoneme, and (5) a uniform distribution of glutamylated as well as glycylated isoforms along the rest of the axoneme. Thus, our data indicate that tubulin molecular heterogeneity is much lower in the Asphondylia axoneme than in the conventional 9+2 axoneme with regard to both isoform content and isoform distribution along the axoneme. Cell Motil. Cytoskeleton 58:160–174, 2004. © 2004 Wiley-Liss, Inc.

Summary Helicobacter pylori is the causative agent of peptic ulcer disease. A major virulence factor of H. pylori is VacA, a toxin that causes massive vacuolization of epithelial cell lines in vitro and gastric epithelial erosion in vivo. Although VacA is exported over the outer membrane and is released from the bacteria, a portion of the toxin remains associated with the bacterial surface. We have found surface-associated toxin to be biologically active and spatially organized into distinct toxin-rich domains on the bacterial surface. Upon bacterial contact with host cells, toxin clusters are transferred directly from the bacterial surface to the host cell surface at the bacteria–cell interface, followed by uptake and intoxication. This contact-dependent transfer of VacA represents a cost-efficient route for delivery of VacA and potentially other bacterial effector molecules to target cells.

A microtubule organizing centre (MTOC) has been described in the spermatid of the hemipteran Matsucoccus feytaudi (Coccoidea). This structure, revealed as a fluorescent ring by treatment with γ-tubulin antibody, gives rise to a bundle of microtubules which surrounds the elongated cylindrical nucleus. This microtubule bundle has been considered an atypical sperm flagellum provided with sperm motility. A comparison of the M. feytaudi MTOC with the material associated with the centriole of Drosophila melanogaster spermatids confirms the great similarity between the two structures, both involved in the nucleation of microtubules. Like the D. melanogaster material associated with the centriole, the M. feytaudi MTOC is a transient structure which disappears or degenerates at the end of spermiogenesis and is no longer visible in the mature sperm.