Abstract
Tardigrades are microinvertebrates inhabiting almost all aquatic and terrestrial ecosystems throughout the world. They are known for their ability to enter into cryptobiosis and to survive extreme environmental conditions (e.g. lack of water, very high and low temperatures, high doses of radiation, vacuum space). Thanks to these abilities, tardigrades are excellent model organisms for various types of studies, e.g. ecological, ethological, physiological, astrobiological, biotechnological or medical, or even in integrative taxonomy. For most of these studies well-established tardigrade cultures are essential. Here we present a review of methods/protocols used in tardigrade culturing in the past. Based on this data and on our several years of experience in tardigrade culturing, we tried different methods and developed new ones that seem to be optimal. Here, we propose our own simple protocols for culturing herbivorous, omnivorous and carnivorous eutardigrade species in environmental chambers as well as in room conditions. We also describe methods for culturing rotifers, nematodes and algae, used as food sources for tardigrades. Moreover, many years of tardigrade culturing allowed us to describe the problems that may occur during culturing, explain their causes and propose solutions. We believe that these simple protocols will be very useful for many scientists planning tardigrade applications in their studies.
Highlights
There is at present no standard method for tardigrade culturing, and such a standard, unified approach for all tardigrade taxa will never be proposed
We describe useful tips and tricks for eutardigrade culturing and discuss possible problems that may arise during such laboratory culturing, as well as propose pos sible solutions
We reared tardigrades at room temperature, which ranged from 21 to 24°C, with relative humid ity (RH) estimated at 40–60% and light conditions naturally regulated in a day/night cycle
Summary
There is at present no standard method for tardigrade culturing, and such a standard, unified approach for all tardigrade taxa will never be proposed. This is due to the fact that different tardigrade species have different envir onmental requirements. The main differences concern the walking substrate, type of food, ambient temperature and photoperiod (for more details see Table I and the Discussion section) Such a diversity of applied protocols, for e.g. culturing of the same species, could be potentially problematic for comparing the results and conclusions of different experi ments focused on different aspects of tardigrade biology. Algae (Pseudochlorella sp.) Nematodes (Pristionchus iheritieri, Panagrolaimus rigidus, Caenorhabditis elegans) Algae Nematodes (C. elegans) Rotifers (L. inermis) and algae (C. vulgaris) Rotifers (L. inermis). We think that the presented protocols can be useful for different types of studies, e.g. ecological, ethological, physiological, astrobiological, biotechnologi cal or medical, or even in integrative taxonomy
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