ECOTOXICOLOGY: ANNELIDS

Drewes C. D., Vining E. P., Callahan C. A. (1984): Non‐invasive electrophysiological monitoring: A sensitive method for detecting sublethal neurotoxicity in earthworms. Environmental Toxicology and Chemistry: An International Journal 3: 599-607. 
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Abstract
Earthworms were exposed, by external surface contact, to three chemical pollutants: dieldrin (a known neurotoxicant), dimethyl phthalate and fluorene (a possible neurotoxicant). After 48 h of exposure, LC50 values were determined and compared with concentrations required for sublethal neurotoxic effects on giant nerve fiber conduction velocity and escape reflex function, as assessed by noninvasive electrophysiological recordings. For dieldrin, sublethal decreases in giant fiber conduction velocity occurred at concentrations as low as 1.4 ppm, whereas much higher concentrations were required for lethality (48-h LC50 = 8,100 ppm). Other sublethal effects of dieldrin included hyperresponsiveness of giant fibers, reduction in giant fiber-mediated muscle potentials, tonic spasms of longitudinal shortening and ataxia. For fluorene, sublethal neurophysiological effects, including reduced conduction velocity and occasional “rebound” spiking of giant fibers, were evident at 100 and 250 ppm. By comparison, the 48-h LC50 for fluorene was 360 ppm. No sublethal neurophysiological effects were evident following treatment with dimethyl phthalate; the 48-h LC50 was 10,900 ppm. The results indicate that noninvasive electrophysiological recording of earthworm escape reflex function can be used as a sensitive means of detecting potential sublethal neurotoxic effects of pesticides and other environmental pollutants in these nontarget organisms.

Eyambe G. S., Goven A. J., Fitzpatrick L. C., Venables B. J., Cooper E. L. (1991): A non-invasive technique for sequential collection of earthworm (Lumbricus terrestris) leukocytes during subchronic immunotoxicity studies. Laboratory Animals 25: 61-67.
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Abstract
We have compared two methods for collecting earthworm leukocytes (coelomocytes) with respect to cell yield, viability and behaviour in immunoassays. Non-invasive extrusion was more efficient than puncturing the coelomic cavity. Extrusion does not produce trauma to earthworms maintained under long term laboratory conditions. Neither technique modified immune assays, as determined by erythrocyte and secretory rosette formation, and phagocytosis, since all 3 were functionally equivalent. After an initial extrusion, sequential leukocyte collections by extrusion are possible at intervals of 6 weeks without affecting total and differential cell counts and rosette formation.

Patricia C. S., Nerea G. V., Erik U., Elena S. M., Eider B., Manu S. (2017): Responses to silver nanoparticles and silver nitrate in a battery of biomarkers measured in coelomocytes and in target tissues of Eisenia fetida earthworms. Ecotoxicology and Environmental Safety 141: 57-63.
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Abstract
The current use and development of applications with silver nanoparticles (Ag NPs) could lead to potential inputs of these NPs to soils. Consequently, it is crucial to understand the ecotoxicological risks posed by Ag NPs in the terrestrial compartment. In the present investigation, the effects produced by PVP-PEI coated Ag NPs were assessed in Eisenia fetida earthworms in comparison with the soluble form (AgNO3). Earthworms were exposed for 1, 3 and 14 days to a range of sublethal concentrations of Ag (0, 0.05 and 50 mg/kg) and at each exposure time, apart from mortality and weight loss of individuals, metallothionein (MT) protein concentration and catalase (CAT) activity were quantified in earthworm tissues. In addition, cellular and molecular level endpoints (cell viability, absolute and relative trophic indices and transcription levels of catalase-cat- and metallothionein-mt-) were measured in coelomocytes extruded from exposed earthworms. Despite the lack of effects in traditional endpoints (mortality and weight loss), Ag NPs and AgNO3 posed changes at lower levels of biological complexity (biochemical, cellular and molecular levels). Both Ag forms induced similar changes in the metal detoxification mechanism (MT, mt) and in the antioxidant response system (CAT, cat) of E. fetida. In contrast, Ag form dependant cytotoxicity and subpopulation ratio alterations (eleocytes/amoebocytes) were recorded in extruded coelomocytes. Complementarily, the use of coelomocytes to assess molecular level endpoints represented a relevant alternative for development of non-invasive biomarkers.

Ečimović S., Grgić M., Bošnjaković R., Velki M. (2019): Biomarker responses in earthworm coelomocyte extract – Noninvasively collected sample for pesticide effect assessment. Chemosphere 234: 837-844. 
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Abstract
Earthworms are often used as model organisms in ecotoxicological research because of their natural habitat where they can be exposed to many different pollutants, including pesticides. Since a number of them has to be sacrificed for sample collection, it would be useful to develop non-invasive methods and techniques suitable for the analysis of target parameters. The aim of this study is to determine whether the coelomocyte extract, obtained by the non-invasive method, can be used to measure responses of biochemical biomarkers and to establish if it can be used in assessing the effects of pesticides already known to have a negative impact on the earthworms. In the present study Eisenia andrei earthworms were exposed for 48 h to organophosphates dimethoate and pirimiphos-methyl using the filter paper contact test. Following exposure, coelomocyte extracts were prepared and acetylcholinesterase (AChE) and carboxylesterase (CES) activities were measured. The percentage of inhibition of the measured enzymes in the coelomocyte extract was compared with the inhibition of the same enzyme activities in the samples obtained from the whole body homogenate. AChE and CES inhibition was observed at all concentrations for both pesticides in different types of samples. Compared to the coelomocyte extract, the level of AChE inhibition was slightly stronger in the whole body homogenate. Inhibition of CES at the same concentrations in different types of samples did not always coincide, especially in the case of dimethoate, however significant inhibition of CES in coelomocyte extract was recorded. This study indicates the possibility of using the coelomocyte extract for measurement of biochemical biomarkers and assessment of pesticide effects.