Objective The study aim was to explore the effect of feeding microenvironment on laboratory animals. Macroenvironment and microenvironment parameters were compared under the dynamic operation of a specific pathogen free laboratory animal room. Methods An isothermal body surface heat transfer model and a tray moisture and ammonia model were established in rabbits, and the laboratory animal room environment under static and dynamic operation were compared and analyzed using computational fluid dynamic simulation. Results The dynamic environment of the laboratory animal room under dynamic operation was not obvious, but the air flow of the microenvironment changed, resulting in heat dissipation, moisture dissipation, and exclusion of pollutants. Some microenvironment parameters may exceed the standard. Conclusions Changes in environmental factors under static and dynamic operation were observed. Enhanced control of the breeding microenvironment status could ensure the welfare and quality of laboratory animals.