A new and sensitive method for measuring in vivo and in vitro cytotoxicity in earthworm coelomocytes by flow cytometry.

This study describes a new and sensitive method for measuring the in vivo and in vitro cytotoxicity of 2 earthworm species, Eisenia andrei and Perionyx excavatus , exposed to copper. Specifically, we measured the number of coelomocyte cells that were affected by copper following in viv o and in vitro exposure by flow cytometry, after calcein acetoxymethyl ester (calcein-AM) staining. We found that the coelomocyte viability of both earthworm species was noticeably reduced in the in vivo cytotoxicity test at concentrations of 100 mg/kg copper in dry soil. However, pathological symptoms, such as mucous secretion and bleeding, swelling, thinning, and fragmentation, and burrowing symptoms were not evident following exposure to copper levels of <400 mg/kg dry soil. In conclusion, the present study demonstrates that calcein-AM is a more sensitive test of earthworm coelomocyte cytotoxicity compared to the traditional individual level toxicity test. Therefore, this test could be used to detect low levels of metal contamination in soils. [ABSTRACT FROM AUTHOR]