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1. The conservation of small aquatic habitats is fundamental to preserving diversity in Mediterranean freshwater ecosystems. Amphibians are particularly endangered, as their chemical world, represented by both abiotic and biotic scent trails, is highly susceptible to environmental changes. 2. Breeding site fidelity in the northern spectacled salamander, Salamandrina perspicillata, an Italian endemic vertebrate, was investigated with respect to the ability of this species to locate its own home water rather than that from other places. The choice of aquatic habitat for spawning was investigated in dark conditions by comparing different types of experimental water (home breeding water vs. breeding water of allopatric populations vs. mineral water vs. distilled water). Fifty-eight reproductive females were collected at four breeding sites in the Lepini Mountains (Latium, central Italy). The number of eggs spawned by each specimen in the different types of experimental water was counted. 3. The very large majority of the eggs were found in the home breeding water. When the eggs counted in the other types of water were compared, no differences were found. 4. These findings revealed that in the northern spectacled salamander the choice of spawning site was affected by the scent trail of its own aquatic habitat. This new information has improved knowledge of the auto-ecology of S. perspicillata, thus contributing to strategies for its conservation. Copyright © 2007 John Wiley & Sons, Ltd.