Ligand arsenic complexation and immunoperoxidase detection of metallothionein in the earthworm Lumbricus rubellus inhabiting arsenic-rich soil.

Although earthworms have been found to inhabit arsenic-rich soils in the U.K., the mode of arsenic detoxification is currently unknown. Biochemical analyses and subcellular localization studies have indicated that As3+-thiol complexes may be involved; however, it is not known whether arsenic is capable of inducing the expression of metallothionein (MT) in earthworms. The specific aims of this paper were (a) to detect and gain an atomic characterization of ligand complexing by X-ray absorption spectrometry (XAS), and (b) to employ a polyclonal antibody raised against an earthworm MT isoform (w-MT2) to detect and localize the metalloprotein by immunoperoxidase histochemistry in the tissues of earthworms sampled from arsenic-rich soil. Data suggested that the proportion of arsenate to sulfur-bound species varies within specific earthworm tissues. Although some arsenic appeared to be in the form of arsenobetaine, the arsenic within the chlorogogenous tissue was predominantly coordinated with S in the form of -SH groups. This suggests the presence of an As::MT complex. Indeed, MT was detectable with a distinctly localized tissue and cellular distribution. While MT was not detectable in the surface epithelium or in the body wall musculature, immunoperoxidase histochemistry identified the presence of MT in chloragocytes around blood vessels, within the typhlosolar fold, and in the peri-intestinal region. Focal immunostaining was also detectable in a cohort of cells in the intestinal wall. The results of this study support the hypothesis that arsenic induces MT expression and is sequestered by the metalloprotein in certain target cells and tissues.

Resistance to copper toxicity in populations of the earthworms Lumbricus rubellus and Dendrodrilus rubidus from contaminated mine wastes.

Two arsenic and heavy metal-contaminated mine spoil sites, at Carrock Fell, Cumbria, United Kingdom, and Devon Great Consols Mine, Devon, United Kingdom, have been found to support populations of the earthworms Lumbricus rubellus Hoffmeister and Dendrodrilus rubidus (Savigny). Lumbricus rubellus and D. rubidus collected from the Devon site and an uncontaminated site were kept for 28 d in uncontaminated soil and in soil containing 750 mg/kg CuCl2, the state of the specimens being recorded using a semiquantitative assessment of earthworm health (condition index). The condition index remained high for all specimens except those of L. rubellus and D. rubidus from uncontaminated sites, which displayed 100% mortality. Bioavailability of Cu in the soils from one uncontaminated and two contaminated sites and in the uncontaminated soil treated with CuCl2 was determined using sequential extraction. Soils from Devon Great Consols had the greatest availability of Cu, Carrock Fell the lowest. Total tissue Cu for L. rubellus and D. rubidus from the contaminated sites did not change significantly for each species during the experiment. Total tissue concentrations of Cu for L. rubellus and D. rubidus from uncontaminated sites increased significantly during the first 7 d, after which mortality was 90%, making it impossible to continue the analysis.

Bioavailability of nonextractable (bound) pesticide residues to earthworms.

There is an ongoing debate regarding whether nonextractable (bound) pesticide residues in soils are occluded or may remain bioavailable in the long term in the environment. This study investigated the release of 14C-labeled residues, which were previously nonextractable after exhaustive extraction with organic solvents in soils, and their uptake by earthworms (Aporrectodea longa). After a 100-day incubation of soils treated with 14C-labeled atrazine, isoproturon, and dicamba and exhaustive Soxhlet extractions with methanol and dichloromethane, nonextracted 14C-labeled residues remaining in the soils were 18, 70, and 67%, respectively. Adding clean soil in the ratio of 7:1 increased the volumes of these extracted soils. After earthworms had lived in these previously extracted soils for 28 days, 0.02-0.2% of previously bound 14C activity was absorbed into the earthworm tissue. Uptake by earthworms was found to be 2-10 times higher in soils containing freshly introduced 14C-labeled pesticides as compared to soils containing nonextractable 14C-labeled residues. The differential bioavailability observed between freshly introduced 14C-labeled pesticides and those previously nonextractable may be related to the ease of transfer of the 14C activity into the solution phase. By the end of the 28-day incubation period, 3, 23, and 24% of previously nonextractable 14C-labeled isoproturon, dicamba, and atrazine residues, respectively, were extracted by solvents or mineralized to 14CO2. The amounts of 14C activity released were not significantly different in the presence or in the absence of earthworms in soils containing previously nonextractable residues. However, the formation of bound residues was 2, 2, and 4 times lower for freshly introduced 14C-labeled isoproturon, dicamba, and atrazine, respectively, suggesting that the presence of earthworms retarded bound residue formation.

Fate of ciliates in the earthworm gut: An in vitro study.

The behavior of the ciliateColpidium campylum was unaffected by exposure in vitro to high concentrations of pharyngeal and hindgut fluids from the earthwormLumbricus terrestris, but movement became abnormal and ceased in concentrations of midgut fluid as low as 1.5%. On mixingC. campylum culture with an equal volume of midgut fluid, which often contained resident astomatous ciliates, the.C. campylum cells were immediately immobilized and frequently disintegrated, while the astomatous ciliates continued to behave normally. Possible causes of the hostility of the midgut environment, and implications for earthworm nutrition, are discussed.