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The present study investigated behavioural responses of naïve rainbow trout Oncorhynchus mykiss, lake charr Salvelinus namaycush, lake whitefish Coregonus clupeaformis and goldfish Carassius auratus to food extracts as well as selected amino acids (at 10−6 M) and related chemicals in a flow‐through behaviour trough. Cysteine, the most potent olfactory stimulating amino acid determined electrophysiologically, enhanced locomotor activity, an initial arousal behaviour, in all species examined, in exactly the same fashion as did food extracts. The increased locomotor activity was followed by distinct species‐specific search behavioural patterns: 1) bottom searching in rainbow trout; 2) surfacing and jumping in lake charr; 3) exploratory behaviour against the trough window in lake whitefish; 4) gravel pecking in goldfish. Olfactory‐stimulating neutral amino acids alanine and serine and gustatory‐stimulating imino acid proline were effective, to varying degrees, in eliciting feeding behaviour. Arginine and glutamate suppressed the locomotor activity in rainbow trout, but triggered feeding behaviour in the other species. Agmatine, a decarboxylated arginine, stimulated feeding behaviour in all species tested, except rainbow trout. The observed behavioural responses were in close accordance with the electrophysiological threshold and specificity. Together, the present data demonstrate that single amino acids and closely related chemicals initiate feeding behaviour primarily by olfaction, and interchangeably and complemented by gustation, in naive fishes.