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President Volodymyr Zelensky has fired the head of Ukraine's security agency (SBU) and the prosecutor general, citing many cases of treason in the two powerful organisations.
He said more than 60 former employees were now working against Ukraine in Russian-occupied areas.
A total of 651 collaboration and treason cases had been opened against law enforcement officials, he added.
The sacked officials, Ivan Bakanov and Iryna Venediktova, have not commented.
In his video address late on Sunday, Mr Zelensky said: "Such an array of crimes against the foundations of the national security of the state... pose very serious questions to the relevant heads [of the two organisations].
"Each of these questions will receive a proper answer," the Ukrainian president added.
The sacking of SBU chief Ivan Bakanov, a childhood friend of Mr Zelensky's, follows the high-profile arrest of a former SBU regional head in Crimea, annexed by Russia in 2014. Oleh Kulinych is suspected of treason.
In other developments on Sunday:
- Ukraine says a significant number of Russian warships in the Black Sea were moved from Crimea further east to the port of Novorossiysk. This comes as Kyiv has received further deliveries of longer-range missile systems from its Western allies
- Dozens of distraught relatives and local residents attended the funeral of four-year-old Liza, who was one of 24 people killed when Russian missiles hit the central Ukrainian city of Vinnytsia on 14 July
- Russia's former President Dmitry Medvedev, now deputy head of the Security Council, warned Ukraine of a "Judgement Day" if it were to attack Crimea