A total of 64.84 per cent of voters said "yes" to the question of whether they wanted to amend the constitution to include a definition of marriage as a "union between a woman and a man", according to partial results from around one-third of polling stations released by the electoral commission.
Croatia's current constitution does not define marriage.
A total of 34.56 per cent of voters said "no", the results showed.
However, three hours before voting ended, the turnout was a rather low 26.75 per cent, the electoral commission said.
Under Croatian law, a referendum does not require a majority voter turnout to be valid.
The centre-left government, rights activists and prominent public figures have all spoken out against the measure.
But the recent unveiling of a government bill enabling gay couples to register as "life partners" sparked fears among conservatives in Croatia -- which joined the EU in July -- that same-sex marriage would be next.
"Marriage is a union of a woman and a man designed so that children are born in it. I'm not certain that a same-sex couple can have children in a natural way," he added ironically.
In May, a Church-backed group called "In the Name of the Family" collected almost 700,000 signatures seeking a nationwide vote on the definition of marriage.
"Marriage is the basis of family and family is the basis of any society, including Croatian," the leader of the initiative, Zeljka Markic, told AFP.
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