To determine the influence of feeding frequency on apparent digestibility and blood metabolites in horses, four geldings were fed a complete ration either once (at 08:00 h) or three times a day (at 08:00, 13:00 and 18:00 h). Horses were provided with a mixture of cereals and chopped alfalfa hay at maintenance level for energy supply, 344 kJ/kg BW(0.75) on a daily basis. After three weeks' adaptation, total amounts of faeces and urine were collected for five days, using collection harnesses. Serial blood samples were taken at -30, -15, 0, 5, 15, 30, 45, 60, 90, 120, 180, 240, 300, 360, 480 and 600 min after feeding and analysed for glucose, L-lactate, triglycerides (TG), non-esterified free fatty acids (NEFA) and triiodothyronine (T(3)). Apparent digestibility coefficients of dry matter (DM), crude protein (CP), ether extract (EE), crude fibre (CF), ash and nitrogen-free extract (NFE) were not significantly different (P>0.05) between the two feeding frequencies. Basal plasma glucose, plasma glucose curves and other plasma blood metabolites were not affected by meal frequency. The horses fed the whole ration in one morning feeding did not consume the entire meal at once, but spared feed for ingestion throughout the day, which may explain the lack of effect. Feeding frequency did not affect plasma glucose response (area under curve :AUC) (P=0.705), but AUC (glucose) was negatively correlated with CP (R(2)=0.76; P=0.005) and CF digestibility (R(2)=0.61; P=0.022). Further research is needed to clarify whether different endocrine responses or differences in passage rate can explain these correlations.