In January, the city's government said it would terminate its contract with the Macao Jockey Club in April.
The decision came at the request of the Macao Horse Race Company, which cited operational challenges as part of the reasons for the closure.
On Saturday, gamblers congregated in the half-full stands and placed their final bets.
Some tourists also visited the track.
Horse racing in Macau has struggled with economic challenges in recent years. (AP PHOTO)
Mai Wan-zun, a student from mainland China in Macau, said she wanted to get a taste of the atmosphere.
"We could come to see horse racing here in Macau but not in mainland China," she said.
Helena Chong, a Macau resident, decided to visit the race course for the first and last time to see what it is all about.
"It's a pity to see the end of all this gambling and entertainment," she said.
Horse racing in the former Portuguese colony has struggled with economic challenges in recent years and has yet to rebound from the effects of the COVID-19 pandemic.
Its jockey club had accumulated operating losses of more than $US311 million ($A477 million), the Macau News Agency earlier reported.
Under the termination arrangement, the horse racing firm had pledged to arrange for transportation of owners' horses to other locations by March 2025 and handle the company's employees according to the law, the government said.
In neighbouring Hong Kong, horse-racing remains popular and profitable.
Its jockey club runs various gambling activities and is the city's major donor of many charity works.
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