Some of the clinical signs seen in horses during endurance races may result from increases in neuromuscular excitability and are related to plasma [K þ ] and [Ca þ þ ]. The present study aimed to test the following hypotheses:(1) Potassium supplementation will affect plasma [K þ ] and may result in clinical signs related to neuromuscular hyperexcitability during an 80 km endurance ride.(2) Plasma [Ca þ þ ] will reflect dietary cation-anion balance (DCAB) and calcium intake. Feeding with a high DCAB and high dietary calcium content (1.5% total calcium of daily ration) diets would lead to higher plasma [Ca þ þ ] during an endurance race than on feeding high DCAB diets with a moderate dietary calcium content (1% of total calcium of daily ration). The current study was undertaken during the 80 km endurance research ride in 2002 in Virginia, USA. Forty volunteer rider-horse pairs participated in the race. During the race, electrolyte mixtures with (EM þ K) and without (EM 2 K) potassium were supplied to 18 and 22 horses, respectively. After the race, the horses receiving EM 2 K during the race were supplied with a recovery formula containing potassium (EM-REC). The horses were fed in addition to their own forage (hay and pasture) either their own commercial concentrate (CC; 1% calcium, n ¼ 11) or one of two research-supplied concentrates during 3 months preceding the research ride, one concentrate rich in sugar and starch (SS; 2% calcium, n ¼ 15) and the other rich in fat and fibre (FF; 2% calcium, n ¼ 14). Peripheral blood samples were taken the day before, within 3 min of the arrival at the vet checks at 27, 48 and 80 km, and after 3 h of recovery. Plasma samples were analysed for pH, haematocrit (Hct), [Na þ ], [K þ ], [Cl 2 ], [Ca þ þ ], [Mg þ þ ], total protein (TP) and albumin [alb]. Effects of sampling times, treatments and interactions were evaluated by ANOVA in a mixed model with repeated measures and applied to the 25 horses that completed 80 km. Eliminated horses had their blood sampled before entering the elimination vet check and 3 h after elimination, and were compared with finishing horses by t-test. As the ride progressed, significant increases were found in plasma pH, [Na þ ], ½PO 2 4 , [TP], [alb], Hct and osmolality; and decreases in [K þ ], [Mg þ þ ], PCO 2 , [Ca þ þ ] and [Cl 2 ]. Horses supplied with potassium-free, sodium-rich electrolyte formulae (EM 2 K) had 12.5% lower (P ¼ 0.001) mean plasma [K þ ], 7.8% lower (P ¼ 0.024) TP and 8.4% lower (P ¼ 0.004) albumin at 80 km, and at 3 h after the race they had 6.8% lower (P ¼ 0.045) TP, when compared with EM þ K supplemented horses. Horses fed with SS and FF had higher [Ca þ þ ] at 27 (P ¼ 0.027), 56 (P ¼ 0.006) and 80 km (P ¼ 0.022) when compared with horses fed with CC. The lower [K þ ] in the EM 2 K group, and the higher [Ca þ þ ] in the SS-and FF-supplemented horses may help prevent increases in neuromuscular excitability and related clinical signs. The lower TP and albumin indicate less dehydration in the EM 2 K group and could help prevent re...