Abstract
The immunotoxicological effects of polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/Fs) mixtures on <em>Eisenia andrei </em>earthworms have never been studied. In this work we investigated these effects both for <em>in vitro </em>and <em>in vivo</em> exposure, using the viability and the phagocytic activity of coelomocytes as immunological biomarkers and the flow cytometry was used for analysis. The <em>in vitro </em>exposure revealed a cytotoxic effect of PCDD/Fs mixture (C2) containing 50&yen;10-3 ng/mL of 2, 3, 7, 8-TCDD and an induction of the phagocytic capacity at the mixture (C1) containing 25&yen;10-3 ng/mL of 2, 3, 7, 8-TCDD. In the <em>in vivo </em>filter paper exposure, the immunocompetence of earthworms was assessed after 3 h-exposure to mixtures of PCDD/Fs at the levels of C1, C2, C3 and C4 containing about; 0.05, 0.3, 0.5 and 0.83 ng of 2, 3, 7, 8-TCDD/cm&sup2;, respectively. Morphological observations showed an excessive secretion of mucus and body surface lesions in worms exposed to higher concentrations (C3 and C4), which revealed that these organisms were affected by PCDD/Fs either through skin and/or by feeding. The levels of the extruded cell yield decreased significantly at all the concentrations tested. However, the cell viability was shown to be unaffected by PCDD/Fs concentrations. It was also shown, that exposure to the highest PCDD/Fs concentrations; C2, C3 and C4 inhibited both phagocytic activity and efficiency.
Highlights
Ly Abstract on The immunotoxicological effects of polychlorinated dibenzo-p-dioxins/dibenzofurans e (PCDDs/Fs) mixtures on Eisenia andrei earths worms have never been studied
Earthworms have been extensively used as competence of oligochaete Eisenia andrei standard test organisms in soil toxicity test- using the pagocytosis assay as immunological Cell extrusion ing[1]
The results of the in vitro exposure showed that C2 presents a cytotoxic effect on cœlomocytes (Figure 2)
Summary
The mixture of PCDDs/Fs in nonane solution was obtained from Wellington laboratories (Ontario, Canada) and stored at 1°C. Earthworms have been extensively used as competence of oligochaete Eisenia andrei standard test organisms in soil toxicity test- using the pagocytosis assay as immunological Cell extrusion ing[1]. Total number of cells and initial cell viability were determined by diluting 20 mL of cell suspension with 20 mL of 0.4% trypan blue (Sigma-Aldrich) and this cell suspension was taining the cell suspension were exposed to dilutions of a PCDDs/Fs mixture; C1 and C2 (Table 2). These dilutions were done in DMSO (final concentration 0.004% per tube).
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