复合乳酸菌制剂的耐受性和抑菌性研究
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摘要
本试验将四株饲用乳酸菌,制成复合乳酸菌制剂,通过单因素试验设计,以存活率为指标,研究复合微生态产品对高温、酸、胆盐的耐受性,采用琼脂稀释法、牛津杯法分别测定复合乳酸菌制剂对不同种类饲用抗生素的耐受性及对致病菌的抑制作用。结果表明:复合乳酸菌产品在温度65℃处理2.5、5、7.5 min以及85℃处理2.5 min对其存活率没有影响,但当85℃处理5 min,存活率下降至48%;p H为3时处理90 min存活率高达80.54%。对0.3%猪胆盐的耐受性较好,处理1 h存活率为73.52%;对抗生素(药物饲料添加剂)杆菌肽锌、亚甲基水杨酸杆菌肽耐受性最强,对吉他霉素的耐受性最差;可抑制致病菌株大肠埃希氏菌的生长。体外评价试验结果表明,复合乳酸菌制剂具有很好的益生特性,可进一步研究其在动物上的应用效果。
In this experiment,four strains Lactic acid bacteria were made into compounded Lactobacillus preparation.The survival rate was used as the evaluation indexes,the in vitro probiotic effects of heat resistance,acid resistance,salt tolerance and bacteriostatic ability,antibiotic resistance were evaluated by single factor test.The results showed that the survival rate reached over 96% under the treatment at the temperature of 65 ℃ for2.5,5,7.5 min,85 ℃ for 2.5 min. But it decreased to 48%when the temperature was increased to 85 ℃ for 5 min.The survival rate reached over 80% under the treatment at the pH of 3 for 90 min.Compound Lactobacillus preparation had the ability to tolerance of 0.3% pig bile salt, the survival rate reached 73.52% for 1 h.The compound Lactobacillus preparation has the strongest tolerance to the bacitracin zinc and bacitracin methyleue disalicylate, and the worst tolerance to kitasamycin;The compound Lactobacillus preparation could inhibit the growth of pathogenic strain Escherichia coli.The vitro probiotic effect indicated that compound Lactobacillus preparation has probiotic properties and good application value.
In this experiment,four strains Lactic acid bacteria were made into compounded Lactobacillus preparation.The survival rate was used as the evaluation indexes,the in vitro probiotic effects of heat resistance,acid resistance,salt tolerance and bacteriostatic ability,antibiotic resistance were evaluated by single factor test.The results showed that the survival rate reached over 96% under the treatment at the temperature of 65 ℃ for2.5,5,7.5 min,85 ℃ for 2.5 min. But it decreased to 48%when the temperature was increased to 85 ℃ for 5 min.The survival rate reached over 80% under the treatment at the pH of 3 for 90 min.Compound Lactobacillus preparation had the ability to tolerance of 0.3% pig bile salt, the survival rate reached 73.52% for 1 h.The compound Lactobacillus preparation has the strongest tolerance to the bacitracin zinc and bacitracin methyleue disalicylate, and the worst tolerance to kitasamycin;The compound Lactobacillus preparation could inhibit the growth of pathogenic strain Escherichia coli.The vitro probiotic effect indicated that compound Lactobacillus preparation has probiotic properties and good application value.
引文
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[23] Hyronim B,Lemarrec C,Sassi A H,et al.Acid and bile tolerance of spore-forming lactic acid bacteria[J].International Journal of Food Microbiology,2000,61(2/3):193~197.
[24] Li Y X,Zhang X L,Li W,et al.The residues and environmental risks of multiple veterinary antibiotics in animal feces[J].Environ Monit Assess,2013,185(3):2211~2220.
[25] Wasson K,Criley J M,Clabaugh M B,et al.Therapeutic efficacy of oral Lactobacillus preparation for antibiotic associated enteritis in guinea pigs[J].Contemporary Topics in Lab oratory Animal Science,2000,39(1):32~38.
[26] Zhang S H,Cao G W,Xu D F,et al.Isolation identification and bacteriostasis of lactobacillus from piglet’s feces[J].Feed Industry,2007,28(14):31~33.
[2]董晓丽,张乃锋,周盟,等.一株乳酸菌GF103的分离鉴定及体外益生效果评价[J].动物营养学报,2012,24(9):1832~1838.
[3]龚慧.猪源益生芽孢杆菌的筛选及发酵工艺研究[D].洛阳:河南科技大学,2014.
[4]顾瑞霞,谭东兴,郭久和.胆汁酸盐和低p H值对乳酸菌活性的影响[J].微生物学通报,1996,23(3):144~146.
[5]韩庆功,崔艳红,王元元.植物乳杆菌的生理特性及体外益生效果研究[J].粮食与饲料工业,2018,3:42~46.
[6]胡生梅,陈应国.微生物学检验[M].江苏:江苏凤凰科学技术出版社,2015:320~325.
[7]李舒宁.猪源发酵乳杆菌的分离筛选、特性及其应用研究[D].兰州:甘肃农业大学,2014.
[8]凌代文.乳酸细菌分类鉴定及试验方法[M].北京:中国轻工业出版社,1993.
[9]刘超齐,王平,常娟.益生菌对温度、pH及抗生素耐受性的研究[J].饲料研究,2016.12,19~25.
[10]刘冬梅,李理,杨晓泉.用牛津杯法测定益生菌的抑菌活力[J].食品研究与开发,2006,27(3):110~111.
[11]刘明珠.乳酸粪肠球菌耐受性、抑菌作用及其对瘤胃发酵功能的影响研究[D].南昌:江西农业大学,2013.
[12]罗睿.鸡源乳酸菌的分离筛选及主要生物学特性研究[D].雅安:四川农业大学,2011.
[13]孙笑非.枯草芽孢杆菌对温度、pH及抗生素耐受性的研究[J].饲料研究,2009.8,66~67.
[14]王彦萍,熊涛,黄涛.优良饲用乳酸菌的筛选及在模拟消化环境中的耐受性[J].食品与发酵工业,2016,42(10):56~60.
[15]吴慧芬,毛胜勇,姚文,等.2株源乳酸菌对低pH值和胆盐耐受性及热稳定性研究[J].华中农业大学学报,2005,24(3):265~268.
[16]杨波.饲用抗生素的使用现状及面临的挑战[J].饲料工业,2007,22:3~5.
[17]杨玲.乳酸菌在动物生产中的应用研究[J].饲料工业,2014,29~31.
[18]张璐.饲用芽孢杆菌产品益生特性的体外评价研究[D].北京:中国农业科学院,2008.
[19]张宜凤.耐热乳酸菌的筛选及乳扇生产工艺条件的研究[D].扬州:扬州大学,2008.
[20]中华人民共和国农业部.关于发布《药物饲料添加剂品种目录及使用规范》的公告(征求意见稿)[Z].2017.11.2.
[21] Collado M C,Sanz Y.Method for direct selection of potentially probiotic Bifidobacterium strains from human feces based on their acid-adaptation ability[J].Journal of Microbiological Methods,2006,66(3):560~563.
[22] Gilliland S E,Staley T E,Bush L.Importance of bile tolerance of Lact Acidophilus used as a dietary adjunct[J].Journal of Dairy Science,1984,67,3045~3051.
[23] Hyronim B,Lemarrec C,Sassi A H,et al.Acid and bile tolerance of spore-forming lactic acid bacteria[J].International Journal of Food Microbiology,2000,61(2/3):193~197.
[24] Li Y X,Zhang X L,Li W,et al.The residues and environmental risks of multiple veterinary antibiotics in animal feces[J].Environ Monit Assess,2013,185(3):2211~2220.
[25] Wasson K,Criley J M,Clabaugh M B,et al.Therapeutic efficacy of oral Lactobacillus preparation for antibiotic associated enteritis in guinea pigs[J].Contemporary Topics in Lab oratory Animal Science,2000,39(1):32~38.
[26] Zhang S H,Cao G W,Xu D F,et al.Isolation identification and bacteriostasis of lactobacillus from piglet’s feces[J].Feed Industry,2007,28(14):31~33.