A study was undertaken in cattle to evaluate changes in milk L-lactate in relation to mastitis. A healthy, rear quarter of the udder of each of ten cows in mid-lactation was infused with 1000 colony-forming units (cfu) of Streptococcus uberis following an afternoon milking. Foremilk samples were taken at each milking from control and treated quarters and antibiotic treatment was applied following the onset of clinical mastitis or after 72 h. One cow did not become infected. Six quarters showed clinical symptoms of mastitis within 24–40 h and this was associated with a more than 30-fold increase in milk L-lactate (to 3·3 mM) and an increase in somatic cell count (SCC) from 4·5×103 to 1×107 cells/ml. Three cows were subclinical, with cell counts ranging from 1·5×106 to 1×107 cells/ml. In these animals, milk lactate ranged from 0·7 to 1·5 mM in the infected quarters up to 40 h post-infection, compared with less than 0·1 mM in control quarters. Milk was examined from 137 cows in mid-lactation which were known to have mastitis. Foremilk samples were taken aseptically from control and infected quarters of cows on commercial farms. Mean milk L-lactate concentrations and SCC were 0·14±0·02 mM and 1·85±0·3×105 cells/ml, respectively, in control (bacteriologically negative) samples. However, L-lactate concentrations exceeded 2·5 mM in the presence of some types of infection, the level of the lactate response being closely related to the impact of the infection on SCC. L-Lactate concentrations were relatively elevated in milk samples taken post partum, declining from 0·8 to 0·14 mM over the first few days of lactation. In conclusion, milk L-lactate has potential as an indicator of clinical and subclinical mastitis in dairy cows.