Role of artificial burrows in Hexagenia toxicity tests: Recommendations for protocol development

Abstract
Hexagenia are an important component of fish and waterfowl diets, provide an ecological link in the conversion of detritus to usable nutrients and are useful test organisms for monitoring trends in aquatic contaminants. Consequently, Hexagenia bilineata were used in toxicity tests to determine their sensitivity to leachate from spent shale oil and to evaluate the influence of including artificial glass burrows in exposure chambers. Gill beat frequency and mortality were significantly higher (p = 0.01) in nymphs that were not afforded an opportunity to burrow than in those exposed to the toxicant but that had access to artificial burrows. Molting frequency was depressed in Hexagenia lacking burrows, and it was further decreased as the amount of toxicant increased. Thus, thigmotactic stress accentuated by the presence of shale oil leachate was relieved by including artificial burrows in the exposure chambers. The resulting toxicity data are more ecologically meaningful because the burrowing life history characteristic of the Hexagenia was addressed and incorporated into the test protocol.