The Serb National Council (SNV), an umbrella organisation for Croatian Serbs, registered 331 cases of violence, hate speech and other forms of discrimination against the ethnic minority in 2016.
The figure was up nearly 60 percent from the previous year.
Ethnic Serb leader and MP Milorad Pupovac labelled it an "eruption of hate... And creation of anti-Serb feeling" and warned that the cases were not isolated.
"There are hundreds of others" which are not registered, he told reporters as presented the council's report.
Ethnic Serbs are Croatia's largest minority, accounting for around four percent of the population of 4.2 million.
Respect of their rights was among the key criteria for Croatia's membership of the European Union in 2013.
But ethnic Serbs warn that anti-discriminatory legislation was mostly not being implemented.
"We miss a public, political reaction ... That will be understandable to all those speaking in that (hate) language," Pupovac said.
He said that the government of conservative Prime Minister Andrej Plenkovic had inherited such a climate.
Plenkovic, who came to power after a snap election in October, has pledged to move away from extremism.
But critics say the current administration has not done enough to move away from its predecessor's policies.
In one such incident of concern last month, posters showing bodies hanging from a tree with the caption "Serbian family tree", written in English, appeared in the eastern town of Vukovar.
The government condemned them as shameful and a 19-year-old man was arrested.
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