The strike started at 8:30 am and ran until the end of service, with a break between 5:00 pm and 8:00 pm.
Meanwhile, Rome’s Barberini and Repubblica stations on the Line A remained closed for repair works, causing further delays and disruption.
The latter station is expected to reopen on Wednesday, after being closed for 364 days.
Traffic was heavier than usual on Tuesday, while delays and cancellations caused confusion and chaos among locals and tourists in the Italian capital.
This strike was called to protest the health and safety conditions of ATAC staff, who claim they’re forced to work in dangerous conditions.
The USB trade union said staff are exposed to frequent attacks and the alleged presence of “lethal” fine dust in the air in the tunnels and stations on Rome’s underground network.
The strike had originally been scheduled for 20 June but was postponed by five days.