Mastitis is one of the most common and costly diseases in dairy cattle. The causative agents of mammary gland infections are mainly bacteria and therefore antibiotics are the first choice for mastitis treatment. Due to increasing antimicrobial resistance and legislative restrictions, there is a need to look for alternative non-antibiotic treatments for mastitis. Options include phytotherapy (plant extracts), cytokines, nanoparticles, enzymes and bacteriocins, which have antibacterial effects. Some of these methods can already be used on Czech farms, other substances have positive results in laboratory conditions, but their effectiveness in practice is still the subject of intensive research.