The oyster mushroom cultivation house typically has multiple layers of growing shelves that cause the disturbance of air circulation inside the mushroom house. Due to this instability in the internal environment, growth distinction occurs according to the area of the growing shelves. It is known that minimal air circulation around the mushroom cap facilitates the metabolism of mushrooms and improves their quality. For the purpose of this study, a CFD analysis FLUENT R16 has been carried out to improve the internal environment uniformity of the oyster mushroom cultivation house. It is found that installing a section of the working passage towards the ceiling is to maintain the internal environment uniformity of the oyster mushroom cultivation house. When all the environment control equipment – including a unit cooler, an inlet fan, an outlet fan, an air circulation fan, and a humidifier – were operated simultaneously, the reported Root Mean Square (RMS) valuation the growing shelves were as follows: velocity 23.86%, temperature 6.08%, and humidity 2.72%. However, when only a unit cooler and an air circulation fan operated, improved RMS values on the growing shelves were reported as follows: velocity 23.54%, temperature 0.51%, and humidity 0.41%. Therefore, in order to maintain the internal environment uniformity of the mushroom cultivation house, it is essential to reduce the overall operating time of the inlet fan, outlet fan, and humidifier, while simultaneously appropriately manage the internal environment by using a unit cooler and an air circulation fan.