Salmonid fishes, native to the northern hemisphere, have become naturalized in many austral countries and appear linked to the decline of native fishes, particularly galaxiids. However, a lack of baseline information and the potential for confounding anthropogenic stressors have led to uncertainty regarding the association between salmonid invasions and galaxiid declines, especially in lakes, as these have been much less studied than streams. We surveyed 25 lakes in the Aysén region of Chilean Patagonia, including both uninvaded and salmonid-invaded lakes. Abundance indices (AI) of Galaxias platei and salmonids (Salmo trutta and Oncorhynchus mykiss) were calculated using capture-per-unit-effort data from gillnets, minnow traps, and electrofishing. We also measured additional environmental variables, including deforestation, lake morphometrics, altitude, and hydrological position (i.e., lake order). An information-theoretic approach to explaining the AI of G. platei revealed that by far the strongest effect was a negative association with the AI of salmonids. Lake order was also important, and using structural equation modeling, we show that this is an indirect effect naturally constraining the salmonid invasion success in Patagonia. Supporting this conclusion, an analysis of an independent data set from 106 mountain lakes in western Canada showed that introduced salmonids are indeed less successful in low-order lakes. Reproductive failure due to insufficient spawning habitat and harsh environmental conditions could be the cause of these limits to salmonid success. The existence of this effect in Chilean Patagonia suggests that low-order lakes are likely to provide natural ecological refugia for G. platei. Finally, pristine, high-order lakes should be actively protected as these have become rare and irreplaceable unspoiled references of the most diverse, natural lake ecosystems in Patagonia.