We investigated the influence of acclimation on results of in situ bioassays with cutthroat trout in metal-contaminated streams. Cutthroat trout (Oncorhynchus clarki) were held for 21 days (1) in live containers at a reference or "clean" site having dissolved metals near detection limits (0.01 ??g/L cadmium [Cd] and 2.8 ??g/L zinc [Zn]; hardness 32 mg/L as CaCO3) and (2) at a site in a mining-impacted watershed having moderately increased metals (0.07 ??g/L Cd and 38 to 40 ??g/L Zn; hardness 50 mg/L as CaCO3). The 96-hour survival of each treatment group was then tested in situ at five sites from September 5 to 9, 2002, and each group exhibited a range of metal concentrations (0.44 to 39 ??g/L arsenic [As], 0.01 to 2.2 ??g/L Cd, and 0.49 to 856 ??g/L Zn). Survival was 100% at three sites for both treatments. However, a higher percentage of metal-acclimated fish survived at the site with the second highest concentrations of Cd and Zn (0.90 and 238 ??g/L, respectively) compared with fish acclimated at the reference site (100% vs. 55%, respectively). Survival was 65% for acclimated fish and 0% for metal-nai??ve fish at the site with the largest metal concentrations (2.2 ??g/L Cd and 856 ??g/L Zn). Water collected from the site with the largest concentrations of dissolved metals (on October 30, 2002) was used in a laboratory serial dilution to determine 96-hour LC50 values. The 96-hour LC50 estimates of nai??ve fish during the in situ and laboratory experiments were similar (0.60 ??g Cd/L and 226 ??g Zn/L for in situ and 0.64 ??g Cd/L and 201 ??g Zn/L for laboratory serial dilutions). However, mortality of nai??ve cutthroat trout tested under laboratory conditions was more rapid in dilutions of 100%, 75%, and 38% site water than in situ experiments. ?? 2007 Springer Science+Business Media, LLC.