To date, there is no reported standard mating ratio for Nigerian native chickens despite their large population in the country. This study investigated the effect of mating ratio on behaviour, semen quality and level of stress. Sixty‐three (9 cocks and 54 hens) sexually matured birds were randomly assigned to three mating ratios (MR) of 1 cock to 3 hens (1C:3H), 1 cock to 6 hens (1C:6H) and 1 cock to 9 hens (1C:9H), and each MR was replicated three times. Each cock spent 2 weeks in a particular MR before it was moved to another. By the 6th week, all cocks had experienced the three mating ratios. Behaviour of the hens and sexual behaviour of the cocks were observed in the morning and evening, respectively, for 6 weeks. At the end of 2nd week in each MR, semen samples were collected and evaluated for motility, morphology, liveability and concentration, and blood samples were also taken from the cocks to determine the heterophil/lymphocyte (H:L) ratio, an indicator of stress. There was also no significant effect of mating ratio on the fertility of eggs and hatchability of the chicks. The number of hen–hen aggression instances, frequency of visits to the nest box and duration of nesting was not influenced by the mating ratio. However, the duration of nest seeking was higher at 1C:3H and lower at 1C:9H. A mating ratio of 1C:9H is therefore recommended for farmers interested in raising Nigerian native chickens.