"The significant increase of ceasefire violations in the Donetsk region represents an unprecedented level of violence" since the latest truce agreement in 2015, the office of EU foreign policy chief Federica Mogherini said in a statement.
The Organisation for Security and Cooperation in Europe (OSCE) said late on Saturday that a team of its monitors carrying out an observation mandate in eastern Ukraine had come under fire 50 kilometres (30 miles) south of rebel hub Donetsk.
There was no damage to the vehicle or people, and the patrol left the area immediately, it said. The OSCE also said it had observed a "significantly higher number of ceasefire violations" in the Donetsk region.
In a separate incident on Thursday, a patrol of monitors in rebel territory observed a bullet hole in their vehicle, but it was not clear precisely when the car was hit.
The incidents of targeting monitors are "unacceptable," the EU statement said, adding that "a sustainable ceasefire is urgently needed."
More than 9,200 people have been killed since the Moscow-backed insurgency erupted in eastern Ukraine in April 2014, a month after Russia annexed the Crimean peninsula, with the rebels now controlling parts of Ukraine's Donetsk and Lugansk regions in the east of the country bordering Russia.
A series of periodic truce deals in 2015 have abated some of the violence, which Kiev and its Western allies accuse Moscow of stirring, but casualties around the line of contact continue to be reported on a regular basis.
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