Objective: This study aimed to determine the appropriate supplementation level of lactic acid bacteria (LAB; Lactobacillus plantarum and Bacillus clausii), yeast (Saccharomyces cariocanus and Wickerhamomyces anomalus) for degrading free gossypol and glucosinolate in the fermented total mixed ration (TMR) containing cottonseed meal (CSM) or rapeseed meal (RSM), to improve the utilization efficiency of these protein sources. Methods: For LAB, L. plantarum or B. clausii was inoculated at 1.0x10(8), 1.0x10(9), 1.0x10(10), and 1.0x10(11) colony-forming unit (CFU)/kg dry matter (DM), respectively. For yeast, S. cariocanus or W. anomalus was inoculated at 5x106, 5x10(7), 5x10(8), and 5x10(9) CFU/kg DM, respectively. The TMR had 50% moisture and was incubated at 30 degrees C for 48 h. After fermen-tation, the chemical compositions, and the contents of free gossypol and glucosinolate were determined. Results: The results showed that the concentration of free gossypol content was reduced (p<0.05), while that of the crude protein content was increased (p<0.05) in the TMR contain-ing CSM inoculated by B. clausii (1x10(9) CFU/kg DM) or S. cariocanus (5x10(9) CFU/kg DM). Similarly, the content of glucosinolate was lowered (p<0.05) and the crude protein content was increased (p<0.05) in TMR containing RSM inoculated with B. clausii (1x10(10) CFU/kg DM) or S. cariocanus (5x10(9) CFU/g DM). Conclusion: This study confirmed that inclusion of B. clausii with 1.0x10(9) or 1.0x10(10) CFU/kg DM, or S. cariocanus (5x10(9) CFU/kg DM) to TMR containing CSM/RSM improved the nutritional value and decreased the contents of anti-nutritional factors.