Trans-galactosyl-oligosaccharides (TOS) are a class of oligosaccharides produced by transgalactosylation of lactose. TOS are used as bifidogenic factors in human and animal nutrition. TOS can be present in the oral cavity and form a risk of caries. All oral bacteria tested were able to degrade and ferment both TOS and galactosyllactose (GLL), one of its components. Growth was improved compared with carbohydrate-free media and acid was produced after 24 h incubation of the bacteria with TOS and GLL. Degradation patterns, using HPAEC, showed degradation of most components. GLL was degraded only partially. Rapid acidification was only observed for Streptococcus mutans, resulting in a pH of 5.4 within 30 min. All other strains fermented TOS and GLL only slowly. Plaque formation could not be detected on both substrates. It can be concluded that TOS and GLL present only a small risk of caries formation, unless proven otherwise in animal studies.