Longitudinal in vivo MR imaging of live earthworms

Earthworm (Oligochaeta, Lumbricidae) species are used widely in eco-toxicological tests especially with contaminated soils. These long-term tests are reliable, but a high sample size is needed. Magnetic resonance imaging (MRI) can produce fast, robust, sensitive, and longitudinal morphological results using a small sample size. Performing longitudinal in vivo examinations of earthworms using MRI requires the need for anesthetics to completely avoid earthworm's moving. Our goal was to develop a simple and non-invasive method to anesthetize earthworms for in vivo longitudinal imaging studies. We investigated a number of different anesthesia methods and found that propan-2-ol and its vapor was optimal. We used a commercial sequential nanoScan® PET/MRI system (Mediso Ltd, Hungary, Budapest) to explore feasibility of MR imaging in immobilized earthworms. It was possible to visualize via micro MRI the brain, gastrointestinal tract, seminal vesicles, calciferous gland (Morren gland), and main blood vessels of the circulatory system. Our findings show the possibilities to examine changes in morphology using MRI of certain organs using a reversible, long-term immobilization method.