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The males of Carabus lefebvrei produce sperm bundles of quite variable size as a consequence of a sexual selection via sperm competition. The ultrastructural features of spermatogenesis of the species and the sequence of events leading to sperm bundle formation along the deferent duct lumen revealed the origin and evolution of the apical structure protecting the sperm bundles. Elongated layers of the electron-dense material of variable size and thickness are visible over the microvilli of the distal deferent duct epithelium. They likely correspond to sections through different levels of discoidal secretions. Then, the sperm present in the duct lumen attach to one side of such elongated secreted structures. Later on, these structures bent to take a cap shape and increase their size and thickness by apposition of additional secretions produced by the proximal deferent duct epithelium. These results reconcile the different opinions about the formation of the structures called the cap and the rod (spermatostyle). In our work, we in fact obtained evidence that both these structures have the same origin, even though at the end of their formation they exhibit a different appearance. The reported results indicate that, differently from the spermatostyle, in the initial stages of cap formation, the sperm adhere only to one side of the cap rather than around the whole structure, as it occurs instead along the spermatostyle rod. Under this aspect, the cap structure is similar to the protection present in the sperm of some whirligig beetle.

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.

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 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.

Halyomorpha halys (Heteroptera: Pentatomidae) is an insect pest native to Asia that has spread over the last two decades to most of the North America, parts of South America, Europe and North Africa. Its impact is significant as it can feed on more than 300 host plants, rendering affected fruits and vegetable crops unsellable or of lower quality. Various chemical and biological methods have been used to control this pest, with varying degrees of success. The sterile insect technique (SIT) is a pest control method involving the sterilization of insects via ionizing radiation and their subsequent mass release into the field. In the present contribution, the spermiogenesis of H. halys was studied from an ultrastructural point of view in both irradiated and non-irradiated adult males. In both cases, we observed ultrastructural characteristics typical of hemipteran sperm cells: bridges connecting the mitochondrial derivatives and the axonemal microtubules, the absence of accessory bodies, and the presence of two or three crystalline inclusions within the mitochondrial derivatives, an acrosome composed of tightly packed tubules, and an atypical, plaque-shaped microtubular organizing center (MTOC) in the centriolar region. Moreover, in the same region, we seldom observed the presence of two centrioles in the spermatids, one of which disappeared at a later stage of maturation. This feature is a novelty for insect spermiogenesis. The cysts of irradiated adults were not all uniformly affected by the radiation. However, irradiated cysts sometimes exhibited a general disorganization of sperm arrangement, incomplete divisions of sperm cells resulting in multiple copies of the same organelle within the same cell, failure to reabsorb the cytoplasm, and the lack of axonemes. Finally, rod-shaped viruses or virus-like particles were observed in vasa deferentia independently of irradiation.

The sperm ultrastructure and the male and female genital apparatus of Zorotypus shannoni were examined and documented in detail, mainly using transmission electron microscopy micrographs. The findings suggest an evolutionary trend shared with Z. hubbardi and Z. impolitus . The three species are characterized by enlarged mitochondrial derivatives and related modifications. Giant sperm are probably a synapomorphy of Z. hubbardi and Z. impolitus , whereas an intermediate condition of this feature is found in Z. shannoni. The monophyletic origin of Z. caudelli , Z. magnicaudelli , Z. huxleyi and Z. weidneri is suggested by characteristically modified axonemes. The presence of extra-acrosomal material is also an unusual feature for Zoraptera, but this condition also occurs in the majority of polyneopteran groups. The long and convoluted female spermathecal duct with secretory and duct-forming cells is a constant feature in Zoraptera. The enlarged seminal receptacle suggests an evolutionary link between the male genital structures and the sperm size on one hand, and the size of the female spermatheca on the other. The small and otherwise uniform group Zoraptera exhibits a remarkable variation of sperm types and genital structures, suggesting the impact of different types of selection. It is likely that cryptic female choice plays a major role in shaping the genital apparatus.

Ground beetle (Carabidae) spermatozoa are often aggregated in bundles of different sizes and are covered by a secretory material produced by the deferent duct epithelium. These secretions can have different shapes being either similar to a cap or to a rod (spermatostyle). Through a detailed study of the different regions of the male reproductive systems, we were able to describe the origin of this protective structure. We have obtained evidence that the secretions from the duct epithelium have initially an irregular shape, with the spermatozoa present in the duct lumen adhering to the structure with their heads on one side only. Later on, along their maturation, these structures bend, taking a cap shape. Hence, this process of secretion and successive evolution of the structures is very similar to that occurring in the spermatostyles. In conclusion, we obtained clear evidence that the two models of the sperm bundle protection share the same origin and thus they can be considered as homologous.

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.

The large group of Dytiscidae, with more than 4300 species, is characterized by an extreme variation in male and female genital apparatus, with Hydroporinae exhibiting the greatest variation. Important structures, such as the spermatheca and the spermathecal gland, show a variable configuration in the several taxa. In Scarodytes halensis, different from Stictonectes optatus, the two structures are fused in a single complex organ; each region of this complex, however, exhibits an epithelium with a quite different ultrastructure. The epithelium of the region corresponding to the spermathecal gland consists of secretory and duct-forming cells, while the region homologous to the spermatheca has a simple epithelium. Sc. halensis is also provided with a very long spermathecal duct connecting the bursa copulatrix to the complex organ. This duct shows an epithelium rich of microtubules and junctional structures to support the thick layer of muscles surrounding the duct. A short fertilization duct is present between the complex organ and the common oviduct. The different organization of the genital apparatus in the two species suggests the presence of a different reproductive strategy. 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.

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.