分离筛选性能优良菌株并进行系统分析评价,以期为健康产品开发提供高性能、安全的功能性菌株。从健康长寿老人粪便中分离筛选乳酸菌菌株,应用生理生化特性和DNA序列分析进行菌株鉴定,并通过耐酸、耐胆盐、耐人工胃液、耐人工肠液的耐受性、药敏试验,研究菌株的抗逆性和安全性;以环磷酰胺作为免疫抑制剂对小鼠进行造模,研究菌株体内免疫功能及对肠道菌群的影响。从健康长寿老人粪便中分离鉴定获得一株乳酸杆菌,鉴定为嗜酸乳杆菌,命名为嗜酸乳杆菌PBIL2-003(Lactobacillus acidophilus PBIL2-003)(简称L2-003);L2-003具有较好的耐酸耐胆盐特性,在人工胃液中2 h的存活率达到50%,能够在人工肠液中有效增殖;L2-003菌株仅对复方新诺明和阿米卡星有抗性,对庆大霉素中度敏感,对其余抗生素均敏感;小鼠体内评价结果显示:与模型组相比,中剂量和高剂量的L2-003可显著增强免疫器官指数(P＜0.05)、促进脾淋巴细胞增殖(P＜0.05)、促进机体抗体分泌、增强小鼠NK细胞杀伤活性;碳廓清实验显示,高剂量L2-003可显著增强小鼠单核吞噬细胞活性(P＜0.05);肠道菌群测定结果显示,中剂量L2-003可显著增加小鼠肠道内乳酸菌水平,降低产气荚膜梭菌水平(P＜0.05)。结果表明,从健康长寿老人粪便中分离获得的一株嗜酸乳杆菌PBIL2-003,具有较好的抗逆性和安全性,具有显著增强免疫力和调节肠道菌群的功能。
To obtain functional strains with excellent performance and safety in food development, strains were isolated and screened from the feces of long-lived elderly people and systematically analyzed and evaluated. The lactic acid bacteria isolated from the feces of the healthy and long-lived elderly were identified based on their physiological and biochemical characteristics and DNA sequencing. To determine the stress resistance and safety of the strain, acid, bile salt, artificial gastric juice, artificial intestinal juice tolerance and drug sensitivity were tested. The immunosuppressive mice model was established with cyclophosphamide immunosuppressant, to study the immune function of the strains and its effect on intestinal flora. The isolated strain was identified as Lactobacillus acidophilus, and named as Lactobacillus acidophilus PBIL2-003 (hereinafter referred to as L2-003). The 2-hour survival rate of L2-003 in artificial gastric juice was 50%, and it effectively proliferated in artificial intestinal fluid, which indicated that L2-003 has good acid resistance and bile salt resistance. L2-003 was only resistant to compound sulfamethoxazole and amikacin, moderately sensitive to gentamicin, and sensitive to other antibiotics, which indicated that L2-003 had good safety. The results of animal experiments showed that the medium and high doses of L2-003 significantly enhanced the index of immune organs (P<0.05), promoted the proliferation of splenic lymphocytes (P<0.05), the secretion of body antibodies, and enhance the killing activity of NK cells in mice compared with the model group. High dose of L2-003 significantly enhanced the activity of mononuclear phagocytes in mice (P<0.05); The medium dose of L2-003 significantly increased the content of lactic acid bacteria in the intestine of mice, and reduced the content of Clostridium perfringens (P<0.05). The strain L2-003 isolated from the feces of the healthy and long-lived elderly has good stress resistance and safety, and it also has the function of significantly enhance the immunity and regulate the intestinal flora.
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