Species-specific sensitivity of earthworm coelomocytes to dermal metal (Cd, Cu, Ni, Pb, Zn) exposures: methodological approach

Plytycz, Barbara, Klimeka, Malgorzata, Homaa, Joanna, Mazura, Agnieszka Irena, Kruk, Jerzy and Morgan, Andrew John 2011. Species-specific sensitivity of earthworm coelomocytes to dermal metal (Cd, Cu, Ni, Pb, Zn) exposures: methodological approach. Pedobiologia 54 (Suppl) , S203-S210. 10.1016/j.pedobi.2011.06.002

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Abstract

Earthworm immune-competent cells (coelomocytes) can be adversely affected by soil metal contamination. The aim of the present paper was to perform comparative studies on the coelomocytes of four lumbricid species dermally exposed for 2 days to filter paper soaked with Ni, Cu, Zn, Cd, or Pb chlorides (1 mM). Coelomocytes were expelled by electro-stimulation, counted, and assayed ex vivo by flow cytometry and spectrofluorometry. The coelomocytes of Allolobophora chlorotica and Dendrobaena veneta contain both amoebocytes and riboflavin-storing eleocytes; consequently, they were subjected to assays based on flow cytometric quantification of autofluorescent eleocytes and spectrofluorometric measurement of riboflavin content in coelomocyte lysates. In Al. chlorotica, the number of coelomocytes, the percentage of eleocytes, and the amount of riboflavin were significantly lower in Cu-exposed worms although these cytometric parameters were less affected by Ni, Zn, Cd exposure, and entirely unaffected by Pb exposure. In D. veneta, such cellular effects were also only observed in Cu-exposed worms. The coelomic fluids of Aporrectodea caliginosa and Lumbricus rubellus are constitutionally impoverished of eleocytes; therefore, these species could not be subjected to assays measuring the riboflavin content of these granular cells. Rather, the amoebocytes of Ap. caliginosa and L. rubellus were subjected to flow cytometric measurements of in vitro neutral red uptake (NR). However, the NR uptake assay was technically demanding, requiring a strictly normalized incubation period over all samples to yield useful comparative data. In contrast, the riboflavin content in the coelomocyte lysates of eleocyte-rich species appears to be a robust and convenient immune-function biomarker of environmental stress.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Subjects:	G Geography. Anthropology. Recreation > GE Environmental Sciences Q Science > Q Science (General) Q Science > QH Natural history > QH301 Biology
Uncontrolled Keywords:	Aporrectodea caliginosa; Lumbricus rubellus; Allolobophora chlorotica; Dendrobaena veneta; Coelomocytes; Riboflavin
Additional Information:	from the 9th International Symposium on Earthworm EcologyXalapa, Veracruz, Mexico, 5th – 10th September 2010. Available online 1 September 2011.
Publisher:	Elsevier
ISSN:	0031-4056
Last Modified:	24 Mar 2017 03:16
URI:	https://orca.cardiff.ac.uk/id/eprint/21908

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