All experiments conducted on animals were previously approved by the Ethics Committee on Animal Testing, Federal University of San Carlos. Chow Preparation For eight weeks, the animals received chow prepared weekly, stored and analyzed. Only carbohydrate, protein, lipid and fibre content in chow were analyzed. Every care was taken to ensure

that these diets remained homogeneous during the entire experimental period. The chow was prepared from a commercial chow (NUVILAB, Purina®) which, after milling, had its fibre content adjusted by adding 30% of oat bran (Oat bran Quaker®), or 300 g/Kg of standard commercial chow. The chow was characterized according to the procedures of Cavaglieri [22]. Table 1 demonstrates the chow compositions. Table 1 Nutritional Composition in grams (g) of the chows used. NUTRIENT CONTROL % EXPERIMENTAL Selleckchem SB203580 MS-275 research buy % Protein (g) 18 24.8 17.4 23.5 Fat(g) 4 12.4 4.9 14.8 Carbohydrate(g) 45.5 62.7 45.6 61.6 Total fibers (g) 21.9 – 18.9 – Insoluble fibers (g) 18 82 14.4 76.1 Soluble fibers (g) 3.9 17.8 4.5 23.8 Exercise Protocol The animals were submitted to a 5-day period of adaptation to the liquid environment (5 minutes on the first day, 15 minutes on the second, 30 minutes on the third, 45 minutes on the fourth and 60 minutes on the

fifth), in accordance with Sampaio-Barros [23]. Importantly, the control groups were submitted in contact with water, but did not perform the exercise. This was done to equalize the stress compared to the exercised group. A tank was used to perform the swimming sessions, were made of plastic and did not have places where animals could cling to. This was necessary to achieve constant exercise. The water temperature

was monitored at approximately 28 ± 2°C. After adaptation, the training consisted of 60 minutes of daily swimming, five days per week, for eight weeks, performed in the afternoon between 14:00-17:00. The moderate intensity they used a load of 5% of their body weight strapped to their backs, which corresponds to intensity below the Thiamine-diphosphate kinase point of BIBW2992 inflection of the lactate threshold curve. At the end of eight weeks training, the animals were submitted to the exhaustion test, characterized by being incapable of keeping themselves on the surface of the water [24, 25]. Animal sacrifice and sample collection Immediately after the exhaustion test, the animals were sacrificed by decapitation. During exsanguination, the mixed arteriovenous blood was collected in heparinized tube and chilled on ice. Blood was then spun at 500g for 15 min to obtain plasma for cytokine and corticosterone analyses. In the following order, the liver, soleus and white and red gastrocnemius muscle were collected and stored at -70°C until the time of measurement of hepatic and muscle glycogen. The white and red portion of the gastrocnemius was divided throughout the major colour of muscle fibres.