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Anthropogenic noise has been related to stress in captive animals; despite this there have been few studies on animal welfare assessment in walk-through zoo enclosures. We aimed to investigate the behavioural effects of noise on a male-female pair of two-toed sloths (Choloepus didactylus), housed in a walk-through enclosure in a zoo in the UK. The animals were filmed for 24 h per day, during three days per week, including days with potential low and high flow of visitors, for three weeks. Sound pressure measurement was performed four times each collection day (twice in the morning, once at noon and once in the afternoon), for 15 min per session, using a sound level meter. The number of visitors passing the enclosure during each session was also recorded. The videos were analysed using focal sampling, with continuous recording of behaviour. Correlations between noise and the behaviours expressed during, and in the 24 h after the acoustic recording, were investigated. The number of visitors correlated with the acoustic parameters. At the moment of exposure, higher levels of noise correlated with decreased inactivity, and longer expression of locomotion and maintenance behaviours for the male; the female spent more time inside a box in these moments. During the 24 h hours after exposure to loud noise, the female showed no behavioural changes while the male tended to reduce foraging. The behavioural changes observed in both individuals have already been reported in other species, in response to stressful events. Our study indicates the need for a good acoustic management in walk-through zoo enclosures where sloths are housed.