Abstract Adrenaline and cortisol, the major stress hormones, are known for its direct control on stress response in fish. Likewise, as an important stress modifier hormone, thyroid hormone has also been implicated in stress response of fish. We tested whether the hypothesis on the phenomenon of inter-hormonal interference, a process that explains the hormonal interactions, operates in fish particularly between adrenaline, cortisol and thyroid hormones. To achieve this goal, indices of acid–base, osmotic and metabolic regulations were quantified after adrenaline challenge in propranolol pre-treated air-breathing fish (Anabas testudineus). Short-term adrenaline (10ngg−1) injection for 30min produced a rise in plasma cortisol without affecting plasma T3 and T4. On the contrary, blocking of adrenaline action with a non-selective blocker, propranolol (25ngg−1) for 90min reduced plasma cortisol along with plasma T4 and that indicate a possible interference of these hormones in the absence of adrenaline challenge. Similarly, a reduction in plasma T3 was found after adrenaline challenge in propranolol pre-treated fish and that suggests a functional synergistic interference of adrenaline with T3. Adrenaline challenge in these fish, however, failed to abolish this propranolol effect. The remarkable systemic hypercapnia and acidosis by propranolol pre-treatment were reversed by adrenaline challenge, pointing to a direct action of adrenaline on acid–base indices probably by a mechanism which may not require β-adrenergic receptor systems. Interestingly, the prominent adrenaline-induced hyperglycemia, hyperlactemia and hyperuremea were not altered by propranolol treatment. Similarly, adrenaline challenge promoted and propranolol reduced the osmotic competencies of the gills, kidneys and liver of this fish as evident in the sodium and proton pump activities. The modified physiologic actions of adrenaline and its modified interaction with THs and cortisol in blocked fish indicate an interaction of adrenaline with cortisol and THs. Our physiologic evidences thus support the hypothesis of the phenomenon of inter-hormonal interference.