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Background, Aim, and Scope: Hexanitrohexaazaisowurtzitane (CL-20) is a relatively new energetic compound sharing some degree of structural similarity with hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), a known neurotoxic compound. Previously, we demonstrated using a noninvasive electrophysiological technique that CL-20 was a more potent neurotoxicant than RDX to the earthworm Eisenia fetida. In the present study, we investigated the effect of CL-20 exposure and subsequent recovery on muscarinic acetylcholine receptors (mAChRs) to further define the mechanism of reversible neurotoxicity of CL-20 in E. fetida., Materials and Methods: We used a noninvasive electrophysiological technique to evaluate neurotoxicity in CL-20-treated worms, and then measured how such exposures altered levels of whole-body mAChR in the same animals., Results and Discussion: A good correlation exists between these two types of endpoints. Effect on mAChR levels was most prominent at day 6 of exposure. After 7 days of recovery, both conduction velocity and mAChR were significantly restored. Our results show that sublethal concentrations of CL-20 significantly reduced mAChR levels in a concentration- and duration-dependent manner, which was accompanied with significant decreases in the conduction velocity of the medial and lateral giant nerve fibers. After 7-day post exposure recovery, worms restored both neurochemical (mAChR) and neurophysiological (conduction velocity) endpoints that were reduced during 6-day exposures to CL-20 concentrations from 0.02 to 0.22 microg/cm(2)., Conclusions and Perspectives: Our findings support the idea that CL-20 induced neurotoxic effects are reversible, and suggest that CL-20 neurotoxicity may be mediated through the cholinergic system. Future studies will investigate other neurotransmission systems such as GABA, glutamate, and monoamine. Ion channels in the nerve membrane should be examined to further define the precise mechanisms underlying CL-20 neurotoxicity.

Concentrations of Hg and Se were determined for a total of 125 Common Loon (Gavia immer) eggs collected from lakes in Alberta, Saskatchewan, Ontario, Quebec, New Brunswick and Nova Scotia, Canada between 1972 and 1997. Resulting data were compared to Hg and/or Se concentrations known or suspected to cause reproductive impairment in birds. Organic (methyl) Hg analyses were also performed on a subset of 24 loon eggs. Thirty-nine of 125 eggs had total Hg levels exceeding those (0.6 microg g(-1) ww, or approximately 2.5 microg g(-1) dw) previously reported to be associated with reproductive impairment in common loons (Barr, 1986), and 9 of 125 eggs had Hg concentrations higher than the level associated with reproductive impairment in birds generally 1 microg g(-1) ww; (Thompson, 1996). Selenium concentrations in loon egg samples were less than levels known to cause reproductive impairment in birds. A weak but significant positive correlation was observed between egg-Hg and -Se concentrations (r = 0.511, p < 0.05). On average, methylmercury accounted for about 87% of total Hg in 24 eggs analysed for both total and organic Hg. In this subset of eggs, the relationship between organic (methyl) Hg and Se was significant (r = 0.538, p = 0.007) while that found between inorganic Hg and Se in the same eggs was not significant (r = 0.353, p = 0.091). This relationship was unexpected and was contrary to relationships established for organic and inorganic Hg vs. Se in adult loon liver and kidney tissue (Scheuhammer et al., 1998b).