The aim of this study was to evaluate the effect of temperature and pH on the activity of hydrolytic enzymes in the cecal region of guinea pigs (Cavia porcellus). This is the first report on enzymatic activity in guinea pigs, showing that there is amilolytic, cellulolytic, proteolytic and lipolytic activity. The values of the evaluated activities were in the following ranges: amylolitic activity (0.150-0.499 U / mg protein), cellulolytic (0.013-0.022 U/mg protein), proteolytic (0.104-0.290 U / mg protein) and lipolytic (2.227-4.049 U/mg protein). Both in the cellulolytic, lipolytic and proteolytic activity there was interaction between temperature and pH. The amylolytic activity was only affected by the temperature, evidencing greater enzymatic activity at a higher temperature (40 ° C). The amylolytic activity is stable at any of the three pHs tested in this study. In addition, the amylolytic activity was 10 times higher than the cellulolytic activity.