Simple SummaryEnrichment programmes are used to enhance the wellbeing of captive animals in zoos. We aimed to evaluate the effects of an enrichment programme, which included novel objects and scent stimuli, on one group of adult European wildcats hosted at Parco Natura Viva-Garda Zoological Park, Italy. We assessed the behavioural changes following the introduction of novel objects (blocks and rags) and scent (synthetic F3) via observations over four study periods (baseline, rags, F3 rags, blocks). Our results suggest that the enrichment programme did not substantively affect the behaviour of the zoo-housed wildcats but showed potential for both the introduction of novel manipulative objects (rags) and the administration of semiochemicals (F3). However, further work is needed to better assess the effects of the synthetic F3 on welfare in zoo-housed wildcats.AbstractObjects and semiochemicals may be used as enrichment in zoos. Domestic cats release Fraction 3 of Facial Pheromone (F3) by rubbing the muzzle to convey relational and territorial information. We aimed to evaluate whether and how the introduction of novel objects and scent stimuli could change the behaviour of one group (N = 5 subjects) of adult European wildcats (Felis silvestris silvestris) hosted at Parco Natura Viva-Garda Zoological Park, Italy. We assessed the behavioural changes following the introduction of novel objects (blocks and rags) and scent (synthetic F3) via observations over four experimental conditions (baseline, rags, F3 rags, blocks) using continuous focal animal sampling. Our results showed that no behavioural differences were found between the different conditions and the baseline, except for the condition with blocks when significantly less exploration was observed. Between conditions, wildcats performed significantly less individual explorative, affiliative, and agonistic behaviours, but more individual inactivity, when exposed to rags after F3 administration. Our findings suggest that the enrichment programme did not substantively affect the behaviour of the zoo-housed wildcats. However, the behavioural differences recorded between conditions suggest that, while novel objects introduced as visual stimuli (blocks) do not affect the wildcat behaviour, novel manipulative objects (rags) might impact their behaviour. Moreover, the changes in affiliative and agonistic behaviours displayed during the condition with exposure to rags sprayed with F3 suggest that such semiochemical could play an appeasement role within this study group.