一种复合微生态制剂的制备及在仔猪生产上的初步应用
发布时间：2018-09-14 14:45
【摘要】：为了得到高抗逆性、高生物活性的益生菌菌株,研究其生理特性,以期进一步地在生产上应用,本研究从健康猪的新鲜粪便及肠道内容物、传统食物等来源分离筛选目标益生菌,分离得到两株抗逆性强、具有潜在益生性能的菌株ZZS16-3和菌株LYP06-2,两者都具有较强的抗逆性,在人工胃液、人工肠液、不同浓度的胆盐环境中存活,并保持较高的存活率。细菌的菌落形态观察、染色镜检、生化试验以及16S rDNA序列分析及同源性比较,综合确定菌株ZZS16-3为枯草芽孢杆菌,菌株LYP06-2为植物乳杆菌。枯草芽孢杆菌ZZS16-3能够产生多种酶类,如蛋白酶、脂肪酶、淀粉酶、纤维素酶和木聚糖酶等,并且能够抑制金黄色葡萄球菌,菌株LYP06-2具有产酸的特性,能够抑制大肠杆菌,具有潜在的益生性能。从毒理学角度评价枯草芽孢杆菌ZZS16-3和植物乳杆菌LYP06-2这两种菌的安全性,进行急性毒性试验,发现试验昆明小鼠未出现中毒症状,无小鼠死亡,剖检未见异常,两者的急性毒性试验最大耐受剂量MTD15000 mg/Kg,按照GB15193.3-2003中急性毒性(LD50)剂量分级表,属于无毒级别。30天大鼠实验结果表明,与对照组比较,复合菌液对大鼠生长发育、代谢无不良影响,脏器指数、血常规、血液生化及病理学检查均未见异常。通过单因素分析,确定最佳碳源为酵母浸粉,最佳氮源为明胶,最佳无机盐为CaCl2,通过正交实验分析得到最佳培养基配方为：0.5%酵母浸粉、1.5%明胶、1.5%CaCl2;通过发酵条件分析确定发酵初始pH为7.2,发酵温度37℃,转速为150 r/min。按照1%的接种量接种到装有6L发酵培养基的发酵罐中,初始pH为7.2,培养温度37℃,转速150 r/min,罐压0.05 Mpa,通风比：1：0.8,培养18 h,得到芽孢生成率90%以上,活菌数为3.1×109 CFU/mL的枯草芽孢杆菌发酵液。饲料中添加含有枯草芽孢杆菌ZZS16-3和植物乳杆菌LYP06-2的复合微生态制剂在一定程度上提高了仔猪的生长性能,减少仔猪腹泻,降低料重比。
[Abstract]:In order to obtain probiotic strains with high resistance to stress and high biological activity, the physiological characteristics of probiotics were studied in order to be further applied in production. Two strains of probiotics, ZZS16-3 and LYP06-2, which have potential probiotics, were isolated and screened from traditional food sources. High survival rate was maintained in different concentrations of bile salt. The colony morphology, staining microscopic examination, biochemical test, 16s rDNA sequence analysis and homology comparison showed that ZZS16-3 was Bacillus subtilis and LYP06-2 was Lactobacillus plantarum. Bacillus subtilis ZZS16-3 can produce a variety of enzymes, such as protease, lipase, amylase, cellulase and xylanase, and can inhibit Staphylococcus aureus. Potential probiotics. The safety of Bacillus subtilis (ZZS16-3) and Lactobacillus plantarum (LYP06-2) were evaluated from the perspective of toxicology. The acute toxicity test was carried out in Kunming mice. The maximum tolerated dose (MTD15000 mg/Kg,) of both acute toxicity tests was according to the acute toxicity (LD50) dose classification table in GB15193.3-2003. The results showed that the compound bacteria solution had no adverse effects on the growth and development and metabolism of the rats compared with the control group. Organ index, blood routine, blood biochemical and pathological examination were not abnormal. Through single factor analysis, the optimum carbon source is yeast powder, and the best nitrogen source is gelatin. The optimum inorganic salt was CaCl2,. The best medium was obtained by orthogonal experiment. The optimum medium was: 0.5% yeast extract 1.5% gelatin, 1.5Cl2, the initial pH was 7.2, the fermentation temperature was 37 鈩，
本文编号：2243012
[Abstract]:In order to obtain probiotic strains with high resistance to stress and high biological activity, the physiological characteristics of probiotics were studied in order to be further applied in production. Two strains of probiotics, ZZS16-3 and LYP06-2, which have potential probiotics, were isolated and screened from traditional food sources. High survival rate was maintained in different concentrations of bile salt. The colony morphology, staining microscopic examination, biochemical test, 16s rDNA sequence analysis and homology comparison showed that ZZS16-3 was Bacillus subtilis and LYP06-2 was Lactobacillus plantarum. Bacillus subtilis ZZS16-3 can produce a variety of enzymes, such as protease, lipase, amylase, cellulase and xylanase, and can inhibit Staphylococcus aureus. Potential probiotics. The safety of Bacillus subtilis (ZZS16-3) and Lactobacillus plantarum (LYP06-2) were evaluated from the perspective of toxicology. The acute toxicity test was carried out in Kunming mice. The maximum tolerated dose (MTD15000 mg/Kg,) of both acute toxicity tests was according to the acute toxicity (LD50) dose classification table in GB15193.3-2003. The results showed that the compound bacteria solution had no adverse effects on the growth and development and metabolism of the rats compared with the control group. Organ index, blood routine, blood biochemical and pathological examination were not abnormal. Through single factor analysis, the optimum carbon source is yeast powder, and the best nitrogen source is gelatin. The optimum inorganic salt was CaCl2,. The best medium was obtained by orthogonal experiment. The optimum medium was: 0.5% yeast extract 1.5% gelatin, 1.5Cl2, the initial pH was 7.2, the fermentation temperature was 37 鈩，
本文编号：2243012
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