Reproductive damage and compensation of wild earthworm Metaphire californica from contaminated fields with long-term heavy metal exposure

Reproduction is a significant biological process for organisms responding to environmental stresses, however, little is known about the reproductive strategies of invertebrates under long-term exposure to contaminations. In this study, earthworm Metaphire californica (Kinberg, 1867) from contaminated fields with an increased metal gradient were collected to investigate their reproductive responses. The results showed heavy metals (Cd, Cu, Zn, and Pb) induced histological damage to earthworms' seminal vesicles, including tissue disorders and cavities, and decreases in mature spermatozoa. Sperm morphology analysis indicated deformity rates were up to13.2% (e.g. head swollen or missing) for worms from the most contaminated site, which coincided with DNA damages. Furthermore, the computer-assisted sperm analysis (CASA) system was employed for the evaluation of sperm kinetic traits. Results suggested earthworms exposed to higher contamination showed a lower sperm viability rate but faster sperm velocity after re-exposure with Cd solution (like the curvilinear velocity and straight-line velocity paraments) compared with those from relatively clean sites. The activities of lactate dehydrogenase and sorbitol dehydrogenase showed the highest 32.5% and 12.5% up-regulation respectively with the increased metal gradient. In conclusion, this study elucidated the earthworm reproductive toxicity, underlying reproductive compensation, metal stress-induced damages, and adaptive responses caused by heavy metal exposure, while also providing the possibility of sperm trait analysis (CASA) for related earthworm toxicological studies.