From the Super Mario Wiki
Queen Buzzbees are the rulers of the Buzzbee hive in Donkey Kong 3. Starting from round three, they begin appearing from the hives that Donkey Kong shakes loose. They take two shots of Stanley's bug spray to defeat; the first shot simply turns them blue. Even after the second shot, they split into four small bees that are still harmful to Stanley. Sometimes they are accompanied by Beespy guards that increase their point value significantly.