The Brazilian foreign ministry say Gerardo Antonio Delgado Maldonado will have to leave the country and that the decision was a "swift measure".
Brazil has declared Venezuela's top senior diplomat in the country as persona non grata.
The tit for tat move stripping Gerardo Antonio Delgado Maldonado of his status comes days after Venezuela’s decision to expel Brazil’s own ambassador to Caracas, Ruy Pereira.
Explaining its decision, Venezuela said Brazil had acted illegally in impeaching its former left-wing president Dilma Rousseff for fiscal irregularities.
Relations between the two counties have deteriorated since Brazil's President Michel Temer took office last year.
Venezuelan President Nicolás Maduro described her impeachment as "a right-wing coup".
The head of Venezuela's powerful Constituent Assembly, Delcy Rodriguez, said on Saturday that "diplomatic relations with Brazil will not be restored until the government reinstates the constitutional order it has effectively broken".
The Brazilian government said the move showed "once again the authoritarian nature of President Maduro's administration".
On Saturday, Venezuela also expelled Canada's top diplomat in Caracas , accusing him of interfering in internal affairs.
Canada's foreign ministry retaliated on Monday, announcing that ambassador Wilmer Barrientos Fernández, who was already abroad, would not be allowed to return.
Brazil and Canada have both become outspoken critics of Mr Maduro and his decision to convene a Constituent Assembly, which effectively replaced the opposition-controlled National Assembly.
They accuse his socialist government of harassing the opposition and violating human rights.
Mr Maduro's announcement prompted mass street protests, which killed more than 120 people in four months.
Earlier this year, following in the US footsteps, Canada unveiled sanctions against 52 foreigners it deemed corrupt and accused of human rights violations.
Among those sanctioned were Russian, South Sudanese and Venezuelan officials, including Maduro.
In August, US President Donald Trump's government imposed sweeping financial sanctions on Venezuela, and labelled Maduro "a dictator".
The sanctions drew an angry rebuke from Caracas, with Foreign Minister Jorge Arreaza calling them the "worst aggression" against the country in two centuries.
Maduro himself, however, said that the measures did not intimidate him "for a moment".
The Venezuelan President's six-year term ends in 2019. He is due to run for re-election next year.
Venezuela has one of the world's highest inflation rates and for years has suffered from a shortage of basic goods, including medicines.
The government blames the crisis on an economic blockade led by the United States, as well as a sharp drop in the international price of oil, its main export.