Colistin is being used as a last-resort drug to treat infections caused by multidrug-resistant (MDR) bacteria in humans. In veterinary medicine, colistin has been used for the treatment and prevention of infectious diseases. In the first study of mcr genes by multiplex PCR in healthy pigs from Serbia, we discovered mcr-1 in 4.85% out of 350 fecal samples. The presence of mcr-1 gene was detected on three farms located less than 100 km apart from each other, predominantly in piglet samples. The results point to the necessity of monitoring of colistin resistance and the mcr genes in food producing animals as well as restricting colistin usage on farms.