X-ray microtomography for imaging of developing spiders inside egg cocoons

• X-ray microtomography revealed spider embryo morphology in untouched cocoon.
• Spider embryos in one cocoon differed by one developmental stage.
• Site-dependent embryo malformation or embryogenesis disorders were not found.
• Survival rate after X-ray exposure depended on embryo age and scanning parameters.

Embryogenesis is especially sensitive to external factors. The changes in its course are often used as biomarkers of environmental impact. Since spider embryogenesis takes place inside cocoons, it is crucial to find a reliable tool to analyze this developmental phase with no intrusion into the cocoons. The aim of this study was to verify the efficacy of X-ray microtomography for non-invasive analysis of embryonic morphology and egg quantity in the cocoons of Xerolycosa nemoralis and Agelena labyrinthica from polluted and reference sites. X-ray microtomography slice images as well as 3D images and animations obtained from digital visualization of those slides were used to study the morphology of embryos and egg arrangement in the cocoons. Any disorders in embryogenesis or malformation of embryos in relation to site of origin have not been found, but inside an egg cocoon of X. nemoralis from the polluted site embryos differing form each other by one developmental stage were identified. Egg calculation revealed a K- reproductive strategy of X. nemoralis from polluted sites. Finally, future prospects and benefits, and weaknessess of this method for the study of spider cocoons have presented.