Urban green space can help mitigate the negative impacts of urban living and provide positive effects on citizens' mood, health and well-being. Questions remain, however, as to whether all types of green space are equally beneficial, and if not, what landscape forms or key features optimise the desired benefits. For example, it has been cited that urban landscapes rich in wildlife (high biodiversity) may promote more positive emotions and enhance well-being. This research utilised a mobile phone App, employed to assess people's emotions when they entered any one of 945 green spaces within the city of Sheffield, UK. Emotional responses were correlated to key traits of the individual green spaces, including levels of biodiversity the participant perceived around them. For a subsample of these green spaces, actual levels of biodiversity were assessed through avian and habitat surveys. Results demonstrated strong correlations between levels of avian biodiversity within a green space and human emotional response to that space. Respondents reported being happier in sites with greater avian biodiversity (p < 0.01, r = 0.78) and a greater variety of habitats (p < 0.02, r = 0.72). Relationships were strengthened when emotions were linked to perceptions of overall biodiversity (p < 0.001, r = 0.89). So, when participants thought the site was wildlife rich, they reported more positive emotions, even when actual avian biodiversity levels were not necessarily enhanced. The data strengthens the arguments that nature enhances well-being through positive affect, and that increased 'engagement with nature' may help support human health within urban environments. The results have strong implications for city planning with respect to the design, management and use of city green spaces.