This study aimed at describing the physiological demands and fatigue development during elite male handball matches. Our hypothesis was that players perform multiple high-intensity activities during periods of the game and develop temporary and end-match neuromuscular fatigue. Time-motion analyses and heart rate (HR) recordings were performed in 40 players during 12 competitive matches. Blood samples were collected and sprint, jump and intermittent exercise performance (YYIE2) was assessed for 18 players at baseline conditions and after two competitive matches, and additional blood sampling and testing were performed for 12 of these players during a friendly match. The time spent with high-intensity running (4.4±2.0 to 3.1±1.7%), the frequency of demanding actions (61±5 to 54±6) and the time with HR above 80% HRmax (62±21 to 41±17%), were lowered from the first to the second half. Average blood lactate during the match was 3.6±2.1 (1.3-8.6) mM. Plasma FFA, glycerol, glucose and uric acid increased (p