title: A new and sensitive method for measuring in vivo and in vitro cytotoxicity in earthworm coelomocytes by flow cytometry.
abstract: 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 vivo 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 100mg/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 <400mg/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.
authors.affiliation: Department of Environmental Science, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Jin Il
authors.initials: JI
authors.lastname: Kwak
authors.affiliation: Department of Environmental Science, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Shin Woong
authors.initials: SW
authors.lastname: Kim
authors.affiliation: Department of Environmental Science, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea. Electronic address: anyjoo@konkuk.ac.kr.
authors.forename: Youn-Joo
authors.initials: YJ
authors.lastname: An
mesh_terms: D000818:Animals; D005434:Flow Cytometry; D066298:In Vitro Techniques; D008856:Microscopy, Fluorescence; D009835:Oligochaeta / Q000187:drug effects*; D012989:Soil Pollutants / Q000633:toxicity*; D013045:Species Specificity
chemical_list: D012989:Soil Pollutants
keywords: Calcein-AM; Coelomocytes; Cytotoxicity; Earthworm; Flow cytometry