【摘要】 为了解从云龙石斑鱼(Epinephelus moara♀×Epinephelus lanceolatus♂)肠道分离出的地衣芽孢杆菌GM-2 (Bacillus subtilis GM-2)、枯草芽孢杆菌GM-39 (Bacillus licheniformis GM-39)的生物学特征与益生特性，通过对胃肠液、pH、鱼胆汁的耐受性等试验进行生物学特征分析，并通过抑菌作用、黏附性、产酶活性、体外抗氧化等试验进行益生特性研究。结果显示，二株芽孢杆菌经人工胃肠液、5%鱼胆汁溶液分别处理1～3 h处理后，存活率分别达70%以上、80%以上以及50%；经不同pH生理盐水处理30 min后，二株菌的存活率分别从0逐渐提高到87.83%、88.25%；二株菌对18种抗生素均不产生耐药性，血琼脂平板培养均未出现溶血现象，菌悬液分别对6株指示菌有抑制作用；二株菌的自聚率达到30%以上，疏水率居于30%～50%;GM-39对黏液蛋白的黏附性显著强于GM-2;GM-39的产酶活力均高于GM-2;GM-2的羟基自由基清除能力和还原能力强于GM-39，而GM-39的DPPH自由基、超氧阴离子自由基的清除能力优于GM-2。结果表明：二株芽孢杆菌均具备较好的耐受性、安全性、抑菌性、产酶活力和抗氧化能力，因而二株芽孢杆菌均具有益生菌开发潜能，为其后续作为益生菌制剂的研究提供了科学理论依据。更多还原

【Abstract】 The biological characteristics and probiotic properties of Bacillus subtilis GM-2 and Bacillus licheniformis GM-39 was investigated, which were isolated from the intestine of Yunlong grouper(Epinephelus moara ♀ ×Epinephelus lanceolatus ♂). The biological characteristics were analyzed through the tolerance tests of artificial gastrointestinal fluid,pH, fish bile, hemolysis, and drug sensitivity, and the probiotic properties were studied respectively through the experiments of antibacterial activity, self-aggregation and hydrophobicity, adhesion, enzyme production activity, and antioxidant in vitro. The results showed that the survival rates of these Bacillus strains were more than 70%, 80%, and 50% respectively, after treatment with artificial gastric solution, artificial intestinal solution, and 5% fish bile solution for 0～3 h. After treatment with physiological saline solutions with pH of 1.0, 2.0, 3.0, 4.0 and 5.0 for 30 minutes, the survival rates of the two strains gradually increased from 0 to 87.83% and 88.25%, respectively. No hemolysis was observed in the blood agar plate test. The GM-39 was sensitive to 10 kinds of antibiotic sensitivity discs such as tetracycline and enrofloxacin, and GM-2 was sensitive to 8 kinds of antibiotic sensitivity discs such as tetracycline and 6 kinds of commonly used aquatic drugs such as enrofloxacin. The B. licheniformis GM-39 was sensitive to 10 kinds of antibiotic susceptibility discs such as tetracycline and 5 kinds of commonly used aquatic drugs such as enrofloxacin, and the B.subtilis GM-2 was sensitive to 8 kinds of antibiotic susceptibility discs such as tetracycline and 6 kinds of commonly used aquatic drugs such as enrofloxacin. The bacterial suspensions of these strains showed inhibitory effects on 6 indicator bacteria, such as Streptococcus agalactiae, Aeromonas hydrophila, and Vibrio parahaemolyticus, while the fermentation supernatants only showed inhibitory effects on 3 indicator bacteria, such as Escherichia coli and Staphylococcus aureus.The autoaggregation rates of these strains reached more than 30%, and the hydrophobicity rates were between 30% and 50%. The B. licheniformis GM-39 had a significantly stronger adhesion to mucin protein than B. subtilis GM-2. Both these strains can synthesize cellulase, amylase, protease, and lipase, but the enzyme-producing activity of B. licheniformis GM-39 is higher than that of B. subtilis GM-2. Both strains have the ability to scavenge DPPH free radicals, hydroxyl radicals,superoxide anion radicals, and reduction ability. B. subtilis GM-2 has stronger hydroxyl radical scavenging ability and reduction ability than B. licheniformis GM-39, and GM-39 has better DPPH free radical and superoxide anion radical scavenging ability than GM-2.Both these two bacillus have good tolerance, safety, antibacterial activity, adhesion, enzyme production activity, and antioxidant capacity, among which GM-39 has advantages over GM-2 in tolerance, enzyme production activity, and adhesion. Therefore, B. licheniformis GM-39 and B. subtilis GM-2 have potential for probiotic development, thus providing a scientific theoretical basis for their subsequent research as probiotic preparations.更多还原