Visitors have a significant impact on the indoor climate of buildings housing works of art, and the relationship between the number of visitors, the indoor air quality and the protection of exposed works of art is an important factor in the overall study of the indoor climate of heritage buildings without mechanical ventilation and/or air-conditioning systems. In view of these concerns and the lack of studies on natural ventilation in heritage buildings, this study aims to analyse the performance of natural ventilation in the São Cristóvão Church in Lisbon, Portugal. For the preparation of this study, an analysis of the natural ventilation of this church was carried out by creating a model in the CONTAM software, and the indoor air quality was analysed based on different international standards and guidelines for carbon dioxide levels and air flow rates (ACH). Estimating the current ventilation strategy, an average ACH of 0.75 h−1 was estimated during the time the church is open, and an ACH of 0.15 h−1 was estimated during the time the doors were closed. In a yearly analysis, an average ACH of 0.30 h−1 was obtained. These air exchange values guarantee EN 16798-1 category I air quality for 72% of the year and category II air quality for 18% of the year. Different natural ventilation strategies were analysed: (a) three scenarios exploring different cross ventilation scenarios; (b) a scenario assuming that the church is closed all year round; and (c) a scenario estimating an increase in the number of visitors, giving an idea of the variations in human pollutants and possible consequences. Taking into account the air infiltration and the fact that masses, an occasional situation with a high number of visitors, are always held just before the church closes, it is guaranteed that carbon dioxide levels will never exceed the limit of 350 ppm above the outdoor values imposed by EN-13779, registering a maximum of 291 ppm.
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