Ecotourism is a strategy for biodiversity conservation, but it involves possible negative effects on animal health and welfare. Large predators such as crocodilians are one of the great public attractions sought after for tourist interactions. Interactions with wild animals and humans can hyperstimulate the hypothalamic‐pituitary‐adrenal axis in the short term, in the case of crocodilians showing an increased corticosterone level, which is indicative of physiological stress. Between September and December 2019, we simulated interactions between tourists and Amazonian crocodilians at Anavilhanas National Park in Central Amazonia, Brazil to evaluate the effects of handling and use of photographic flashes on black caiman (Melanosuchus niger) and spectacled caiman (Caiman crocodilus) on circulating corticosterone and lactate. Corticosterone levels increased 1.7‐fold during handling and 2.7‐fold when exposed to photographic flashes in black caiman but not in spectacled caiman. Increased corticosterone concentrations in black caiman were characterized by an increase caused by handling and were more intense after flashes than in controls, but the combination of handling and flash had no effect. During handling in simulated tourist interactions, anaerobic respiration increased lactate in black caiman but not in spectacled caiman. The effect of simulated tourist interactions with Amazonian crocodilians was dependent on the handling and especially on flash use in black caiman. The results can assist management, conservation programs, and public policies, especially in programs based on tourism interaction with Amazonian crocodilians.