Nudibranchs are renowned for their complex ecological and chemical defensive adaptations. How environmental heterogeneity shapes their spatial distribution and secondary metabolite profiles remains poorly understood, particularly in the upwelling region of Arraial do Cabo (Brazil). This study investigated how multi-scale environmental variables - specifically temperature and substrate characteristics - influence nudibranch assemblages and chemical profiles across two sites. Field surveys were integrated with computational analyses and Thin-Layer Chromatography to assess inter- and intra-specific variations. Environmental heterogeneity was found to partially modulate nudibranch abundance. Deviations from micro-habitat correlations led to the formulation of a high mobility hypothesis, suggesting wider home ranges that decouple nudibranchs from their immediate substrate. The mismatch between high crude extraction yields and faint or absent chemical bands in juveniles suggested a higher proportion of primary metabolites that went undetected under the performed mobile phases and dilution. This study delivers a major methodological contribution by establishing an innovative, reproducible sampling and extraction protocol for individual, small-bodied specimens, preserving the resolution necessary to detect fine-scale intra-specific variation in future ecological and chemical investigations. Finally, this work represents a pioneering, multi-scale attempt to systematically cross-reference nudibranch ecology with environmental heterogeneity, ranging from macro-oceanographic gradients down to micro-scale habitat features.
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