Investigation of the spermathecal morphology, reproductive strategy and fate of stored spermatozoa in three important thysanopteran species.

In insects, females can keep sperm capable of fertilisation over a long period with the help of the spermatheca. The effectiveness of storing fertile sperm is expected to reflect in the reproductive strategy and, thus, the morphology of the involved organs. In this work, we focused on the relationship between reproduction and morphology in the haplodiploid Thysanoptera, especially if a loss of these traits occurs under thelytoky. The spermathecal morphology and the fate of stored spermatozoa were studied by microscopic techniques (high-resolution x-ray computed tomography and transmission electron microscopy) in three species with different reproductive modes and lifestyles (Suocerathrips linguis, Echinothrips americanus, Hercinothrips femoralis). Mating experiments were conducted to analyse the use of the transferred sperm in the thelytokous H. femoralis. Results show that the spermathecae are relatively simple, which can be explained by the availability of sperm and the short lifespan of the females. However, the spermatheca in H. femoralis seems to be vestigial compared to the arrhenotokous species and females do not use sperm for fertilisation. No substantial change was observed in the structure of spermatozoa, despite an enlargement of the sperm organelles being measured during storage in all three species. The results of this work demonstrate differences in the morphology of the spermatheca, especially concerning the reproduction mode, promoting the understanding of the complex interaction between morphology and behaviour.

Direct three-dimensional imaging for morphological analysis of electrospun fibers with laboratory-based Zernike X-ray phase-contrast computed tomography.

Electrospinning is a well-established and widely used method for the production of protein-based fibrous biomaterials. The visualization of the morphology and the characterization of sample features related to the three-dimensional (3D) structure, like the porosity and fibers thickness, is crucial for the design and fabrication of tailor-made and application-optimized materials. Here, we evaluated the benefits of using 3D X-ray imaging in a laboratory setup with a resolution in the sub-micrometer range for the characterization of electrospun gelatin fibrous mats. We used phase-contrast X-ray computed tomography at the nanoscale (nano-CT) for the evaluation of the time-course morphological changes of the mats induced by chemical cross-linking of the gelatin fibers. We present an image processing protocol that enables the segmentation of the fibers and quantification of the mats porosity, the analysis of the shape and size of the pores, and of the fibers thickness and orientation. We compared the results obtained from the processed nano-CT data with those obtained with the conventional methods used for the characterization of electrospun fibrous materials, and we discuss the advantages and limitations of each method when applied to gelatin electrospun samples. Our results reveal that the use of phase-contrast nano-CT provides quick additional and relevant information for the characterization of fibrous mats and, thus, provides beneficial insights for the design and fabrication of novel fibrous materials.