Cytology and Cytogenetics

Several cytological aspects have been considered in tardigrades. Firstly, the cell constancy which is not a true eutely being several mitoses present even after hatching, even though some organs, such epidermis and nervous ganglia, have the same cell number in juveniles and adults. The total number of these cells is speciesspecific. Then the ultrastructure of cuticle, epidermis, feeding and digestive apparatus, excretory and osmoregulatory organs, muscles, nerve cells, sensory cells and storage cells has been considered. Instead, the ultrastructure of the germ cells has been considered in the chapter on reproduction. With regard to chromosome number and shape, it has been observed that generally there is little difference among the species (n ¼ 5 or n ¼ 6), but several cases of polyploid populations exist, often very similar to diploid populations from a morphological point of view. In most cases the polyploid populations do not have males and reproduce by apomixis. Studies on the genome size have confirmed the presence of polyploid populations, as well as the presence of nuclei with multiple amounts of DNA within the same specimen. The genome size of the tardigrades is always relatively small and does not seem related to phylogenetic lineages. Studies on tardigrade genomes have placed this phylum at the centre of discussions on the evolution of Metazoa and have considered the role of horizontal gene transfer in animal evolution with contrasting results

Dormancy in Freshwater Tardigrades

For more than two centuries, tardigrades have been well known for their ability to undergo dormancy. However, this capability has been well studied mainly in the so-called limnoterrestrial species, i.e., in the species colonizing moist terrestrial habitats, such as mosses, lichens, and leaf litter. In these kinds of substrates, tardigrades are active only when a film of water is available around their body so in this condition they behave like aquatic animals. When the substrate dries or freezes, tardigrades achieve dormancy (quiescence) by entering cryptobiosis, specifically anhydrobiosis or cryobiosis, respectively. In freshwater habitats, both forms of cryptobiosis have been verified only in species able to live both in freshwater and terrestrial habitats. In the truly freshwater (or limnic) species, anhydrobiosis has not been verified, while cryobiosis has been confirmed in a few species. Another dormancy phenomenon bound to diapause is frequent in freshwater species: encystment (sometimes found even in limnoterrestrial species). The cyst state, which involves deep structural and physiological modifications, has been known from the beginning of the past century, but only recently has its morphology and inducing factors been studied in depth. Although data on molecular mechanisms allowing cryptobiosis are available, this information does not exist for encystment

Reproduction, Development and Life Cycles

In tardigrades reproduction occurs only through eggs, fertilized or unfertilized, and therefore only through gametes. Tardigrades exploit several reproductive modes, amphimixis, self-fertilization and thelytokous parthenogenesis (both apomixis and automixis). These modes are often in close relationship with the colonized environment. As regards sexuality, tardigrades can be gonochoristic (bisexual or unisexual) or hermaphroditic. The anatomy of the reproductive apparatus of males, females and hermaphrodites and the maturative patterns of male and female germinal elements are presented and discussed, as well as the ultrastructure of spermatozoa and eggs, including their phylogenetic implications. In addition, mating and fertilization patterns, embryonic and post-embryonic development, sexual dimorphism and parental care are considered and discussed. Finally, vegetative reproduction does not occur in tardigrades, and their capability to regenerate is limited to a physiological tissue restoration of a few cells

Experimentally induced repeated anhydrobiosis in the Eutardigrade Richtersius coronifer

Tardigrades represent one of the main animal groups with anhydrobiotic capacity at any stage of their life cycle. The ability of tardigrades to survive repeated cycles of anhydrobiosis has rarely been studied but is of interest to understand the factors constraining anhydrobiotic survival. The main objective of this study was to investigate the patterns of survival of the eutardigrade Richtersius coronifer under repeated cycles of desiccation, and the potential effect of repeated desiccation on size, shape and number of storage cells. We also analyzed potential change in body size, gut content and frequency of mitotic storage cells. Specimens were kept under non-cultured conditions and desiccated under controlled relative humidity. After each desiccation cycle 10 specimens were selected for analysis of morphometric characteristics and mitosis. The study demonstrates that tardigrades may survive up to 6 repeated desiccations, with declining survival rates with increased numberof desiccations. We found a significantly higher proportion of animals that were unable to contract properly into a tun stage during the desiccation process at the 5th and 6th desiccations. Also total number of storage cells declined at the 5th and 6th desiccations, while no effect on storage cell size was observed. The frequency of mitotic storage cells tended to decline with higher number of desiccation cycles. Our study shows that the number of consecutive cycles of anhydrobiosis that R. coronifer may undergo is limited, with increased inability for tun formation and energetic constraints as possible causal factors

Environmental Adaptations:Encystment and Cyclomorphosis

Stressful environmental conditions generally limit animal survival, growth, and reproduction and may induce dormancy in the form of various resting stages. Tardigrades represent one of a few animal phyla in which different forms of dormancy are frequently encountered. One of these forms, cryptobiosis, a quick response to sudden changes in the environment, has gained a great deal of attention, whereas much less is known of the slower emerging form of dormancy, diapause. In this review we present the current knowledge of diapause in tardigrades. Diapause in tardigrades, represented by encystement and cyclomorphosis, is likely controlled by exogenous stimuli, such as temperature and oxygen tension, and perhaps also by endogenous stimuli. These stimuli initiate and direct successive phases of deep morphological transformations within the individual. Encystment is characterized by tardigrades that lie dormant\u2014in diapause\u2014within retained cuticular coats (exuvia). The ability to form cysts is likely widespread but presently only confirmed for a limited number of species. In tardigrades, cyclomorphosis was first reported as a characteristic of the marine eutardigrade genus Halobiotus. This phenomenon is characterized by pronounced seasonal morphological changes and in Halobiotus involves stages with an extra protecting cuticle. Cyst formation in moss-dwelling limnic species may also occur as part of a seasonal cyclic event and can thus be viewed as part of a cyclomorphosis. Therefore, whereas diapause generally seems to be an optional response to environmental changes, it may also be an obligate part of the life cycle. The evolution of encystment and cyclomorphosis finds its starting point in the molting process. Both phenomena represent an adaptation to environmental constraints. Notably, the evolution of diapause is not necessarily an alternative to cryptobiosis, and some tardigrades may enter both forms of dormancy. The simultaneous occurrence of several adaptive strategies within tardigrades has largely increased the resistance of these enigmatic animals toward extreme environmental stress