The behavioral effects of alcohol in worms are most often assessed by exposing the animals to alcohol and then tracking their locomotion either on an agar surface (crawling) or in a liquid medium (swimming) (e.g., Alaimo et al., 2012; Davies et al., 2003; Morgan and Sedensky, 1995; Speca et al., 2010). Tissue alcohol concentrations, which are substantially lower than exogenous concentrations, continue to rise slowly over the course of at least 50 minutes of exposure (Alaimo et al., 2012). Additional behavioral assays determine the effects of alcohol on egg-laying or hypercontraction of the body wall muscles (Davies et al., 2003; Hawkins et al., 2015). Worms develop acute functional tolerance to alcohol over the course of a 30-minute continuous exposure, which is observed as a decrease in locomotor sedation caused by alcohol despite increasing tissue alcohol concentrations (e.g., Davies et al., 2004; Jee et al., 2013; Mathies et al., 2015; Raabe et al., 2014). In addition, worms can develop chronic tolerance to alcohol, which is observed by withdrawal-induced clumping behavior after 20 hours of exposure (Davies et al., 2004), by withdrawal-induced
