本实验利用芽孢杆菌发酵棉籽粕，研究对棉粕中游离棉酚降解率的影响，及对发酵前后饲料营养成分如活菌数、中性蛋白酶酶活和酸溶蛋白等的影响，并对发酵效果较好的菌株进行菌株鉴定、安全性分析及发酵工艺探索。研究发现：可高效降解棉籽粕中游离棉酚含量及改善营养品质的芽孢杆菌为BLCC1-0039，游离棉酚的测定方法确定为高效液相色谱法；菌株形态观察和生理生化鉴定并结合其16S rDNA序列，初步确定其为枯草芽孢杆菌(Bacillus subtilis)；枯草芽孢杆菌杆菌BLCC1-0039的体内安全性评价，发现未对实验小鼠造成形态和活动异常，也未观察到脏器器官病变，初步确定了枯草芽孢杆菌杆菌BLCC1-0039的体内安全性；枯草芽孢杆菌BLCC1-0039发酵棉籽粕提高发酵料的酸溶蛋白含量和中性蛋白酶活性并有效降低游离棉酚含量：发酵48 h时，发酵棉籽粕酸溶蛋白为26.96%，中性蛋白酶活性为3897 U/g，游离棉酚降解率为97.81%。

The cottonseed meal fermented by Bacillus was used in this study to investigate the effects on the degradation rate of free gossypol in cottonseed meal, as well as the effects on the nutrient components before and after fermentation, such as viable count bacteria, neutral proteases activity and acid-soluble protein content.. In addition, the strains with better fermentation effects were used for identification, and safety analysis and fermentation techonlogy were also performed. The results showed that the Bacillus subtilis, which could efficiently degrade the free gossypol and improve the nutritional quality of cottonseed meal, was BLCC1-0039, and the free gossypol was determined by high performance liquid chromatography. The BLCC1-0039 was preliminarily identified as the Bacillus subtilis by the morphology, and by the biochemical characteristics combined with 16S rDNA sequence. Thein vivo safety evaluation of B.BLCC1-0039 showed that no morphological and activity abnormalities appeared for the mice used, and no organ lesions were observed, which preliminarily suggested the safety of B.BLCC1-0039. Also, the fermentation induced by B.BLCC1-0039 could effectively reduce the content of free gossypol in cottonseed meal and improve the neutral proteases activity and acid-soluble protein content, the acid soluble protein was 26.96%, the activity of neutral protease was 3897 U/g and the degradation rate of free gossypol was 97.81% after fermentation for 48 h.

泰安市科技发展计划（201540701）