The demonstrators made their way to the Belgian capital to present their demands to the European Commissioner for Agriculture, Janusz Wojciechowski.
Central and Eastern Europe farmers took to the streets in Brussels on Tuesday to protest against Ukrainian grain imports, which they say are affecting their livelihoods.
The group wants the EU to take long-term, decisive measures on the issue, which is causing tensions in the region.
Last year, the EU scrapped import duties on Ukrainian agricultural products to support the embattled country, but the cheap grain has forced many local farmers out of their own markets.
"We need some measures so that the transit will be the transit and not the end of the transport, so we need the grains to leave for poorer countries, to Africa, because we have a lot of grains, to sell our owns," Darie Bizu, a Romanian farmer told Euronews.
The bloc struck a deal with five member states at the end of April allowing them to stop selling and storing Ukrainian grain within their territories, while also allocating them financial support of €100m, but the agreement expires on 5 June.
Farmers are now demanding additional EU funds as well as stronger measures, such as enhanced quality control for imports.
Eastern European producers say they can not compete with Ukrainian ones, since they are not operating under the EU's strict green rules and pesticide directives.
József Luzsi, vice president of the Hungarian Chamber of Agriculture said this is simply not fair.
"A Ukrainian producer does not produce under the same conditions as an EU producer," he said.
"We have green regulations that are mandatory in the EU and the use of chemicals that may be allowed in Ukraine, are forbidden for a long time now in the EU."
Farmers presented their demands to the European Commissioner for Agriculture in Brussels on Tuesday.
At the request of Euronews, a Commission spokesperson said that before taking any further steps they expect all member states to implement the previous agreements and withdraw unilateral import bans, which Hungary has so far failed to do.
"For the moment we are still waiting for the removal of some of the unilateral national measures that were introduced before we follow up on the next steps," Commission spokesperson Miriam Garcia Ferrer said.
Other countries asking for financial aid and facing problems like drought, have warned that help to farmers in Eastern Europe cannot be at the expense of other pressing matters.