The responses of wildlife to environmental factors are of conservation importance. However, the absence of relevant information due to inadequate studies, and lack of understanding of the influences of environmental factors on wildlife, particularly in the Zambezian bioregion, remain a conservation concern. For instance, there is a shortage of knowledge on the relationship between fixed effects of environmental factors and behaviour as well as nest group sizes of Smith’s bush squirrels, Paraxerus cepapi Smith, 1836. Our study examined the relationship between fixed effects of several environmental factors (i.e., with focus on ecological factors) and the behaviour as well as nest group sizes of the bush squirrels in and around Zambia’s Chembe Bird Sanctuary (CBS). Flight initiation distances (FIDs) as an index of bush squirrel behaviour, and nest group sizes were simultaneously surveyed to understand the drivers of biological and population responses, respectively. The results revealed that higher tree height (m) and larger canopy coverage (%) could increase FIDs for bush squirrels. The nest group sizes of bush squirrels could also increase with canopy coverage (%) and presence of termite mounds. By focusing on the two different squirrels' responses, the results of this study highlight the most important environmental factors to consider in minimizing the impacts of human activities on bush squirrels, especially in conservation planning and management by taking into consideration the bush squirrels’ natural history, habitat protection and safe distance between humans and bush squirrels.