The foreign ministry in Madrid has summoned France’s ambassador after French farmers seized Spanish lorries and drained their cargo of wine.
Spain said the attack by the French protesters was a “flagrant violation of various basic principles” of the EU.
On Monday, farmers protesting against unfair competition attacked the lorries at a border crossing.
The farmers blame falling food prices on foreign rivalry, supermarkets and distributors.
Spanish farming organisations say tens of thousands of litres of wine were dumped as five vehicles were targeted near Le Boulou, in southern France.
The Spanish government says it reported the incident to the European Commission.
“These incidents, which are occurring all too frequently, are grounds for concern for the Government of Spain,” the foreign ministry said in a statement.
“Spain has already officially passed on its protest to the French authorities, and urges them to adopt all appropriate measures to guarantee, with absolute security, the free movement of people and goods, one of the basic principles of the European Union.”
A Spanish association representing lorry drivers said French police had not acted to prevent the attacks.
Frederic Rouanet, president of the winemakers’ union of the French region of Aude, vowed to continue the protests.
Protesters were protecting their products from cheap wine coming from Spain and Italy, he told AFP news agency.
In July, Madrid issued a similar protest after French farmers targeted Spanish lorries importing farm produce.