The matatu operators went on rampage to bring light to alleged harassment by traffic police officers.
Bonfires were lit at the Kisumu Bus terminus, as well as the Kondele and Kibuye areas, throwing transport into chaos.
Residents in the lakeside city were forced to walk to their work places as motorcycle operators took advantage of the situation and charged exorbitant fares.
Kisumu Bus Terminus Transport Chairman Aggrey Ouma said they would not relent until traffic officers stop the constant arrest of drivers and conductors.
Ouma claimed that the officers mounted numerous road blocks along the major roads leading into Kisumu town, extorting money from the matatus and making unnecessary arrests.
The Busia, Kakamega and Nairobi routes were all blocked by irate operators, and police had a hard time clearing the road to allow other vehicles into the city.
At the main terminus, police had to retreat from a hail of stones directed thrown at them.
The officers have refused to comment on the stalemate.