A total of 63 more embassy staff are due to depart from the Czech Republic by Monday after the Czech Republic accused Russia of involvement in an explosion at a munitions depot in 2014.
More Russian diplomats and their families flew out of the Czech Republic on Saturday amid a dispute between the two nations over Russian involvement in an ammunition depot explosion in 2014.
In April the Czech Republic ordered 18 Russian diplomats to leave in a tit-for-tat move after Russia expelled Czech diplomats from Moscow over the diplomatic row.
Czech leaders have said two agents from Russia's elite GRU Unit 29155 - the same pair thought to be responsible for the 2018 Salisbury poisonings in the UK - had had a role in the 2014 blast in Vrbetice, which killed two people. Russia has denied any involvement.
The Czech Republic has also said Russia won't be allowed to have more diplomats in Prague than it currently has at its embassy in Moscow.
Some 54 Russian embassy employees and their families boarded a flight out of the Czech Republic today. Nine more are expected to depart on Monday, May 31.