Characterisation by Excitation-Emission Matrix Fluorescence Spectroscopy of Pigments in Mucus Secreted of Earthworm Eisenia foetida Exposed to Lead

The earthworm exposed to toxics shows physiological responses as: avoidance and mucus secretion. Heavy metals are particularly toxic to earthworms and the mucus secretion has been considered as a defence mechanism against undesirable substance. The chromophores present in the mucus secretion of Eisenia foetida have been poorly studied. Mucus secretion of E. foetida was induced by PbCl2. High PbCl2 concentrations provoked abundant mucus secretion which showed fluorescence when illuminated by UV light. Dialysis membrane separation, UV Visible and Excitation-Emission Matrix Fluorescence (EEM) spectroscopy were used to characterise the fluorescent pigments. EEM spectroscopy analysis of the mucus secretion signalled three excitation-emission peaks at: 310/380 nm, 370/520 nm and 440/520 nm. Two fluorophores were separated by dialysis. One of them matched the fluorescent compound riboflavin excitation-emission profile; the other is a protein with a peak 290/350 nm. Native-PAGE electrophoresis was conducted to assess the riboflavin-biding ability of the coelomic fluid protein produced by Eisenia foetida showing a high riboflavin-biding ability.