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The bus left Ekamai bus terminal at 10.15 pm for Trat province with about 50 passengers on board.
But about 30 passengers got off in Chanthaburi with 25 remaining on the bus.
As the bus arrived near a curve which is a slope and in front of Khao Samingwithayakom school, it smashed into a light pole, overturned and fell into the ditch.
The driver and 20 others were injured, including a foreign tourist, 10 in serious condition.
They were later admitted to Khao Saming, Trat and Bangkok hospitals in Trat province.
Khao Saming police said initial investigation revealed drowsy driving might be a cause of the accident as passengers said the bus swaying left and right before smashing into the light pole in the middle of the one-way lane.
The driver, Khampee Manee-orn, was booked for reckless driving causing death to others.