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A mixed‐stock fishery occurs when multiple populations of a fish species are exploited together in a common area where they aggregate outside the breeding season (e.g. for feeding or overwintering), and the aggregation is known as a mixture. Recreational fishing often exploits such mixtures, and estimating the proportional contributions of populations to fisheries promotes more sustainable resource use. Ten DNA microsatellites were assayed in a mixture of bull trout Salvelinus confluentus Suckley overwintering in the Nechako River, upper Fraser River, British Columbia, and in baseline population samples from 14 tributaries putatively contributing to the overwintering mixture. A DNA microsatellite‐based mixed‐stock fishery analysis suggested that five populations together contributed 0.80 to the mixture. Most of the errors associated with the mixture estimates were attributable to uncertainty in baseline allele frequencies. Radiotracking data confirmed that tributary populations contributing to the mixture estimated by genetic analysis also contained individuals that moved between spawning tributaries and overwintering sites. The results better resolve habitat use by potadromous bull trout in the upper Fraser River and, in combination with assessments of baseline population‐specific spawning abundances and productivity, will better inform a decision of whether or not allowing some harvest within the current catch‐and‐release fishery is biologically sustainable.

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