丁酸梭菌优良菌株LXYB-2抑菌活性及抗逆性研究
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摘要
该研究以一株丁酸梭菌(Clostridium butyricum)LXKJ-1为出发菌株,通过常压室温等离子体(ARTP)诱变得到的一株耐丁酸钠突变菌株LXYB-2,采用牛津杯法比较菌株LXYB-2与菌株LXKJ-1的抑菌能力,同时采用活菌计数法对两株菌株的耐高温、人工胃液、人工肠液及胆盐等抗逆性进行比较。结果表明,两株菌株对9种常见致病菌均有较好的抑菌活性,除单核细胞增生李斯特氏菌和金黄色葡萄球菌外,两株菌的抑菌活性差异显著(P<0.05)。在高温90℃处理5 min时,两株菌耐受性差异显著(P<0.05);人工胃、肠液耐受性实验结果无显著差异(P>0.05);在胆盐质量分数达到0.4%时,两株菌胆盐耐受性差异显著(P<0.05)。优良菌株LXYB-2更适合作为益生菌饲料添加剂产品应用于动物养殖业。
Using Clostridium butyricum LXKJ-1 as the starting strain, a mutant strain LXYB-2 with stress resistance to sodium butyrate was obtained by atmospheric pressure plasma at room temperature(ARTP) mutagenesis. The antibacterial activity of strain LXYB-2 and strain LXKJ-1 was compared by cupping method, and the resistance of the two strains to high temperature, artificial gastric juice, artificial intestinal fluid and bile salt was compared by viable count method. The results showed that the two strains had good antibacterial activity against nine common pathogenic bacteria. Except for Listeria monocytogenes and Staphylococcus aureus, the two strains showed significant differences in antibacterial activity(P<0.05). At the high temperature treatment of 90 ℃ for 5 min, the resistance of the two strains was significantly different(P<0.05), there was no significant difference in the resistance to artificial gastric and intestinal fluid(P>0.05). When the bile salt mass fraction reached 0.4%, the tolerance of the two strains to bile salts had a significant difference(P<0.05). The superior strain LXYB-2 was more suitable as a probiotic feed additive for application in animal husbandry.
Using Clostridium butyricum LXKJ-1 as the starting strain, a mutant strain LXYB-2 with stress resistance to sodium butyrate was obtained by atmospheric pressure plasma at room temperature(ARTP) mutagenesis. The antibacterial activity of strain LXYB-2 and strain LXKJ-1 was compared by cupping method, and the resistance of the two strains to high temperature, artificial gastric juice, artificial intestinal fluid and bile salt was compared by viable count method. The results showed that the two strains had good antibacterial activity against nine common pathogenic bacteria. Except for Listeria monocytogenes and Staphylococcus aureus, the two strains showed significant differences in antibacterial activity(P<0.05). At the high temperature treatment of 90 ℃ for 5 min, the resistance of the two strains was significantly different(P<0.05), there was no significant difference in the resistance to artificial gastric and intestinal fluid(P>0.05). When the bile salt mass fraction reached 0.4%, the tolerance of the two strains to bile salts had a significant difference(P<0.05). The superior strain LXYB-2 was more suitable as a probiotic feed additive for application in animal husbandry.
引文
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[4]陆俭,张雪平,孟筱琦.酪酸梭菌和婴儿型双歧杆菌对霍乱弧菌的拮抗试验[J].微生物学通报,2000(5):338-341.
[5]贾丽楠,崔嘉,陈宝江.丁酸梭菌对肉仔鸡饲料加工过程及消化道环境的耐受性研究[J].动物营养学报,2017,29(10):3787-3791.
[6]刘磊,田亚男,王鹤松,等.丁酸梭菌CBM01的碳、氮源优化及其对胃肠道耐受性的研究[J].动物营养学报,2017,29(10):3831-3836.
[7]王腾浩,宗鑫,汪以真,等.产抑菌蛋白的丁酸梭菌的筛选和鉴定及体外益生功能研究[J].中国畜牧杂志,2015,51(13):75-81
[8]曹广添,刘婷婷,周琳,等.丁酸梭菌对致病菌和有益菌的体外作用效果研究[J].中国饲料,2012(4):16-19.
[9]孙亮.Bacillus atrophaeus HAB-1的抑菌活性研究及培养条件优化[D].海口:海南大学,2013.
[10]罗璠,李明雄,杨志荣,等.产类细菌素的乳酸菌的筛选及其抑菌特性研究[J].四川大学学报:自然科学版,2010,47(3):655-660.
[11]张秀红,刘婷婷.具广谱抑菌活性乳酸菌的筛选[J].乳业科学与技术,2010,33(2):56-59.
[12]廖秀冬.丁酸梭菌的筛选及其对动物抗氧化能力和肉鸡肉品质影响的研究[D].北京:中国农业大学,2015.
[13]孔青.丁酸梭菌培养与发酵动力学以及调节腹泻小鼠肠道菌群平衡的研究[D].杭州:浙江大学,2006.
[14]林秋云,杜光文,曾树权,尹映华.潮州市食品中食源性致病菌污染状况及耐药性监测结果[J].中国热带医学,2008(6):1015-1017.
[15]NAKANISHI S,TANAKA M.Sequence analysis of a bacteriocinogenic plasmid of Clostridium butyricum and expression of the bacteriocin gene in Escherichia coli[J].Anaerobe,2010,16(3):253-257.
[16]WANG L Y,HUANG Z L,LI G,et al.Novel mutation breeding method for Streptomyces avermitilis using an atmospheric pressure glow discharge plasma[J].J Appl Microbiol,2009,108(3):851-858.
[17]龚利敏,王恬.饲料加工工艺学[M].北京:中国农业大学出版社,2010:136-151.
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[19]陈桂芳,刘艳,单春乔,等.费氏丙酸杆菌生物学特性及与5株益生菌拮抗性试验研究[J].中国酿造,2018,37(1):69-73.
[20]霍恺.乳杆菌旳益生特性研究[D].济南:山东大学,2015.
[21]付小猛.枯草芽孢杆菌CCAM 080032的益生特性及高密度固态发酵工艺的研究[D].武汉:华中农业大学,2011.
[22]张媛媛.复合芽孢杆菌制剂发酵工艺及其耐受性研究[D].哈尔滨:东北林业大学,2012.