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The aim of this work was to validate a protocol for induction of subacute ruminal acidosis (SARA) (Experiment 1) and test the efficiency of probiotic Saccharomyces cerevisiae or monensin to avoid pH ruminal drops in sheep (Experiment 2). In Experiment 1, six ewes were fasted for two days and then fed most with concentrate during four days. Ewes in this protocol had ruminal fluid pH below 6.0 and kept it for 75 consecutive hours. In Experiment 2, 18 sheep were distributed into three groups: Control (CG, n = 6), monensin (MG, n = 6) and probiotic group (PG, n = 6). SARA was induced according Experiment 1. PG had lower pH (5.7 ± 0.1) than CG (6.0 ± 0.1) (P = 0.05), while MG (5.7 ± 0.1) was similar to both during SARA induction. SARA induction reduced ruminal protozoa population (P < 0.05) and increased chloride concentrations in ruminal fluid (P < 0.01). In serum, SARA increased concentrations of phosphorus (P < 0.01), AST (P < 0.01) and GGT (P < 0.01), but reduced LDH (P < 0.01). In conclusion, the protocol used for SARA induction was able to maintain ruminal pH between 5.5-6.0 for more than 48 hours. However, monensin and probiotics supplementation was not effective in preventing changes in ruminal and serum parameters during SARA.