Surgical staff behavior in operating theatres is one of the factors associated with indoor air quality and surgical site infection risk. The aim of this study was to apply an approach including microbiological, particle, and microclimate parameters during two simulated surgical hip arthroplasties to evaluate the influence of staff behavior on indoor air quality. During the first hip arthroplasty, the surgical team behaved correctly, but in the second operation, behavioral recommendations were not respected. Microbiological contamination was evaluated by active and passive methods. The air velocity, humidity, temperature, and CO2 concentration were also monitored. The highest levels of microbial and particle contamination, as well as the highest variation in the microclimate parameter, were recorded during the surgical operation where the surgical team behaved “incorrectly”. Turbulent air flow ventilation systems appeared more efficient than in the past and very low air microbial contamination was reached when behavior was correct. Therefore, adherence to behavioral recommendations in operating theatres is essential to not undermine the effectiveness of the heating, ventilation, and air conditioning systems and employed resources.