A survey of trout species was carried out to assess the trout stocks' quality inhabiting the Palena River Basin, one of the most important rivers for recreational angling in Chilean Patagonia. Six sampling campaigns were carried out over 16 selected stretches of the river between February 2012 and June 2013. A total of 912 trout were collected, 57% rainbow trout (Oncorhynchus mykiss), 33.3% brown trout (Salmo trutta), and 9.6% brook trout (Salvelinus fontinalis). Also, one tiger trout (Salmo trutta × Salvelinus fontinalis) (a hybrid between brown and brook trout) was recorded. Results showed that both rainbow trout and brown trout shared the same reaches of the river, with very similar feeding habits, while brook trout inhabited smaller and more enclosed streams. Maturity stages (V-VI) were recorded from the fish sampled over the period spring-summer-autumn in the case of rainbow trout; autumn-winter for brook trout, and summer-autumn for brook trout. One of the study's most notable features was the small size of the mature brook trout, reaching 17.5 cm in length for gravid females and 12.4 cm for mature males at age 2+. The maximum age recorded for rainbow and brown trout was 6+, with a maximum length of 69.8 and 58.5cm, respectively, while for brook trout, the maximum age recorded was 3+ for a female with a length of 29.9 cm length and 2+ for a male of 16.6 cm.