Muscle biopsy samples were collected from the left middle gluteal muscle of the horses participating in day 2 (speed and endurance test) of a three day event. Six Thoroughbred horses were biopsied the day before and within 30 minutes of completion of the speed and endurance test. Serial muscle sections were reacted histochemically for myosin adenosine triphosphatase activity after acid pre-incubation to demonstrate Type I, IIA and IIB fibers and the glycogen content in the individual fibers was assessed using the periodic acid Schiff (PAS) reaction. Total glycogen in muscle was measured fluorometrically after hydrolysis to glucose. A significant decrease (P less than 0.001) in total muscle glycogen of 306.0 +/- 34.6 mmoles glucose units/kg (dry weight) (mean +/- SE) occurred when samples collected before exercise were compared to those collected following exercise. A significant decrease (P less than 0.05) in the percentage of Type I fibers having medium PAS staining intensity occurred when pre and post-exercise samples were compared. Significant decreases (P less than 0.01 and P less than 0.001 respectively) in the percentage of Type IIA and IIB fibers classified as having high PAS staining intensity occurred when post-exercise samples were compared with those pre-exercise. There were significant increases (P less than 0.01 and P less than 0.001 respectively) in the percentage of Type IIA and IIB fibers classified as having medium PAS staining intensity when the samples collected after the exercise were compared with those before the exercise.