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The "National March for Life" attracted 50,000 people, according to the Catholic Episcopal Conference of Slovakia which organised the event.
"Man did not give himself life; it is a gift given to him," Stanislav Zvolensky, Archbishop of Bratislava, told marchers in the city centre.
"We want freedom for unborn children so that they can live freely their human life," one of the lay march leaders, Marek Michalcik, told the crowd.
Organisers also called on public officials to actively support institutions focusing on helping families in need as well as pregnant mothers and their children.
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Around 66 percent of the Slovak population of 5.4 million is Catholic.
According to an opinion poll conducted earlier this month by the independent Focus agency, only 11.6 percent of Slovaks want tighter abortion legislation.
As they gear up for the February 2020 general election, the governing Smer Social Democrats and their nationalist SNS coalition partners have abandoned plans to tighten access to abortion by imposing an eight week limit.
The number of abortions in the country has been decreasing.
While in 1997 more than 20,000 women chose to have an abortion, last year only 7,350 did so, according to Slovakia's Statistical Office.