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Rome was home to some of the first public toilets in modern times
There are many cafes and restaurants bordering the walls of the Vatican that have restrooms but they are often for paying customers only. However, sometimes a break for a caffè or spremuta d'arancia (fresh squeezed orange juice) goes well with a bathroom stop, and the €1 for an espresso can be worth it. Just ask the barman, "Dov'è il bagno? (Where's the bathroom?)"
The toilets in and around the Vatican are open from 7:00 in the morning until 18:00 in the evening during the winter months and 19:00 during the summer months. During special events, particularly ones expecting a large influx of people, the Vatican places extra portable toilet facilities around the city.
It is important to remember that not all restrooms in the Vatican or in Italy are accessible to those with disabilities and not all restrooms will have disabled stalls.
This map roughly represents where the public toilets are located. The green symbol represents the outdoor Vatican restrooms. The orange symbols inside the walls represent the restrooms inside the Museums. The purple elephant symbols represent changing tables in the women’s restrooms.