副干酪乳杆菌L1的安全性及益生性评价
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摘要
副干酪乳杆菌L1是分离于自然发酵甘薯酸浆中的优势乳酸菌,进行安全性和益生性评价。本实验通过耐药性实验、有害代谢产物实验及动物实验来评价副干酪乳杆菌L1的安全性,通过对外界环境耐受性及生理功能特性实验,评价该株菌的益生性。结果表明,常见的8种抗生素中该菌对红霉素和阿奇霉素表现为抗性,对青霉素表现为敏感,其他均不敏感且不含质粒;有害代谢产物检测结果均为阴性,也不会产生溶血现象;小鼠在7 d和30 d喂养期间一切生命活动正常,说明该菌不会引起急性毒性和亚慢性毒性;在酸性和胆盐环境下L1的总体活菌数始终在10~7 CFU/mL以上,高盐环境下,L1的总体活菌数始终在10~(10) CFU/mL以上;L1孵育1 h的自聚集性为15.8%±1.2%,L1的疏水性为9.7%±0.8%;对Caco-2细胞黏附率为22.37%±1.44%。由此可知,副干酪乳杆菌L1具有良好的安全性和益生性,为生产安全健康的地瓜淀粉絮凝剂奠定基础并且为实际生产应用提供科学依据。
Lactobacillus paracasei L1 was the dominant Lactobacillus isolated from naturally fermented sweet potato sour slurry and its safety and probiotics were evalualed. The safety of Lactobacillus paracasei L1 was evaluated by drug resistance test,harmful metabolites test and animal experiment. The probiotics of Lactobacillus paracasei L1 was evaluated by environmental tolerance and physiological function test. The results showed that the bacteria was resistant to erythromycin and azithromycin,sensitive to penicillin,insensitive to other antibiotics and without plasmid,the results of toxic metabolites test were negative and hemolysis did not occur,all life activities of mice were normal during 7 and 30 days of feeding,indicating that the bacteria did not cause acute toxicity and subchronic toxicity. The total viable bacteria count of L1 was always above 10~7 CFU/mL in acidic and biliary environments,and the total viable bacteria count of L1 was always above 10~(10) CFU/mL in high salinity environments,the self-aggregation of L1 was 15.8%±1.2% in 1 h incubation,the hydrophobicity of L1 was 9.7%±0.8%,and the adhesion rate to Caco-2 cells was 22.37%±1.44%. It could be concluded that Lactobacillus paracasei L1 has good safety and probiotics,which laid a foundation for the production of safe and healthy flocculant of sweet potato starch and provided scientific basis for practical production and application.
Lactobacillus paracasei L1 was the dominant Lactobacillus isolated from naturally fermented sweet potato sour slurry and its safety and probiotics were evalualed. The safety of Lactobacillus paracasei L1 was evaluated by drug resistance test,harmful metabolites test and animal experiment. The probiotics of Lactobacillus paracasei L1 was evaluated by environmental tolerance and physiological function test. The results showed that the bacteria was resistant to erythromycin and azithromycin,sensitive to penicillin,insensitive to other antibiotics and without plasmid,the results of toxic metabolites test were negative and hemolysis did not occur,all life activities of mice were normal during 7 and 30 days of feeding,indicating that the bacteria did not cause acute toxicity and subchronic toxicity. The total viable bacteria count of L1 was always above 10~7 CFU/mL in acidic and biliary environments,and the total viable bacteria count of L1 was always above 10~(10) CFU/mL in high salinity environments,the self-aggregation of L1 was 15.8%±1.2% in 1 h incubation,the hydrophobicity of L1 was 9.7%±0.8%,and the adhesion rate to Caco-2 cells was 22.37%±1.44%. It could be concluded that Lactobacillus paracasei L1 has good safety and probiotics,which laid a foundation for the production of safe and healthy flocculant of sweet potato starch and provided scientific basis for practical production and application.
引文
[1]Maria Barbara Pisano,Silvia Viale,Stefania Conti,et al.Preliminary evaluation of probiotic properties of Lactobacillus strains isolated from Sardinian dairy products[J].Bio Med Research International,2014,29(10):1913-1922.
[2]Drouault-Holowacz,Bieuvelet S,Burckel S,et al.A double blind randomized controlled trial of a probiotic combination in 100 patients with irritable bowel syndrome[J].Gastroenterol Clin Biol,2008,32(2):147-152.
[3]Ouwehand A C.Antiallergic effects of probiotics[J].Journal of Nutrition,2007,137:794-797.
[4]张刚.乳酸细菌-基础、技术和应用[M].北京:化学工业出版社,2007:33-35.
[5]Pham M,Lemberg D A,Day A S.Probiotics:Sorting the evidence from the myths[J].Medical Journal of Australia,2008,188:304-308.
[6]Aguirre M,Collins M D.Lactic acid bacteria and human clinical infection[J].Journal of Applied Microbiology,1993,75(2):95-107.
[7]张莉力,许云贺,李新华.甘薯酸浆中微生物絮凝性研究[J].食品工业科技,2010(6):172-175.
[8]张莉力,许云贺,李新华.副干酪乳杆菌副干酪亚种L1的培养条件及絮凝活性研究[J].食品工业科技,2011(3):139-145.
[9]沈雪.利用Caco-2细胞研究三种常用乳制品益生菌对肠道屏障功能影响[D].长春:吉林大学,2012.
[10]Yongtao Fei,Li Liu,Dongmei Liu.Investigation on the safety of Lactobacillus amylolyticus L6 and its fermentation properties of tofu whey[J].LWT-Food Science and Technology,2017,84:314-322.
[11]南晓芳.豆豉和腐乳中耐盐乳酸菌的筛选、鉴定及其功能特性研究[D].呼和浩特:内蒙古农业大学,2016.
[12]张灼阳,刘畅,郭晓查.乳酸茵耐药性的研究进展[J].中国微生态学杂志,2007,19(5):478-480.
[13]王梦姣,李少英,韦婉.分离自酸马奶的肠球菌属乳酸菌抗性试验[J].食品科技,2013,38(10):7-11.
[14]Shokryazdan P,Jahromi MF,Liang JB,et al.In vitro assessment of bioactivities of Lactobacillus strains as potential probiotics for humans and chickens[J].Journal of Food Science,2017,82(4):2734-2745.
[15]王梦姣,李少英,李淑芬,等.马奶及其制品中7株肠球菌属乳酸菌的安全性评价[J].食品科学,2014(17):204-208.
[16]Shekh S L,Dave J M,Vyas B R M.Characterization of Lactobacillus plantarum strains for functionality,safety and γ-amino butyric acid production[J].LWT-Food Science and Technology,2016,74:234-241.
[17]Yeo S,Lee S,Park H,et al.Development of putative probiotics as feed additives:Validation in a porcine-specific gastrointestinal tract model[J].Applied Microbiology & Biotechnology,2016,100(23):1-12.
[18]Yakabe T.Safety assessment of Lactobacillus brevis KB290 as a probiotic strain[J].Food and Chemical Toxicology,2009,49:2450-2453.
[19]Nancy J,Szabo.Safety evaluation of Lactobacillus pentosus strain b240[J].Food and Chemical Toxicology,2011,49:251-258.
[20]Tmr DA,Garcia EF,De OAA,et al.In vitro characterization of Lactobacillus strains isolated from fruit processing by-products as potential probiotics[J].Probiotics & Antimicrobial Proteins,2017:1-13.
[21]Lavillalerma L,Pérezpulido R,Martínezbueno M,et al.Characterization of functional,safety,and gut survival related characteristics of Lactobacillus strains isolated from farmhouse goat’s milk cheeses[J].International Journal of Food Microbiology,2013:136-145.
[22]Garcia Estefania F,Luciano Winnie A,Xavier Danilo E,et al.Identification of lactic acid bacteria in fruit pulp processing byproducts and potential probiotic properties of selected Lactobacillus strains[J].Frontiers in Microbiology,2016,7:1371.
[23]Runjun Dowarah,Formal analysis,Investigation.Methodology selection and characterization of probiotic lactic acid bacteria and its impact on growth,nutrient digestibility,health and antioxidant status in weaned piglets[J].Plos One,2018,13(3):e0192978.
[24]Monique S do Carmo,Francisca MF Noronha,Mariana O Arruda.Lactobacillus fermentum ATCC 23271 displays in vitro inhibitory activities against candida spp[J].Frontiers in Microbiology,2016,7:1722.
[25]李姗姗.潜在益生乳杆菌的抗生素敏感性研究[D].保定:河北农业大学,2012.
[26]Clementi F,Aquilanti L.Recent investigations and updated criteria for the assessment of antibiotic resistance in food lactic acid bacteria[J].Anaerobe,2011,17(6):394-398.
[27]周蕊,白瑶,李凤琴.食品工业用益生菌安全性评价研究进展[J].卫生研究,2012,41(3):511-514.
[28]顾旭峰,艾连中,宋馨,等.两株干酪乳杆菌的安全性评价[J].食品与发酵科技,2015,51(2):63-65.
[29]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,1999.
[30]Sanders M E,Walker D C,Walker K M,et al.Performance of commercial cultures in fluid milk applications[J].Journal of Dairy Science,1996,79:943-955.
[31]任大勇.益生乳酸杆菌的黏附及免疫调节作用研究[D].长春:吉林大学,2013.
[32]张俊娟.具有黏附性乳杆菌的筛选与特性研究[D].保定:河北农业大学,2011.
[33]Pablo F,Perez,Minnaard Y,et al.Surface propertiesof bifidobacteria strains of human origin[J].Applied and Environmental Microbiology,1998,64(1):21-26.
[34]王娉婷.传统发酵食品来源的功能性益生菌筛选及其在干酪加工中的应用[D].长春:吉林农业大学,2015.
[35]陈臣.植物乳杆菌ST-Ⅲ对肠上皮细胞的黏附性质及机理的研究[D].无锡:江南大学,2008.
[36]丁素娟,王薇薇,李爱科,等.益生菌肠道黏附性研究进展[J].饲料工业,2016,23(13):55-61.
[37]张莉.植物乳杆菌的黏附特性研究及其在益生菌干酪中的应用[D].长春:吉林大学,2013.
[2]Drouault-Holowacz,Bieuvelet S,Burckel S,et al.A double blind randomized controlled trial of a probiotic combination in 100 patients with irritable bowel syndrome[J].Gastroenterol Clin Biol,2008,32(2):147-152.
[3]Ouwehand A C.Antiallergic effects of probiotics[J].Journal of Nutrition,2007,137:794-797.
[4]张刚.乳酸细菌-基础、技术和应用[M].北京:化学工业出版社,2007:33-35.
[5]Pham M,Lemberg D A,Day A S.Probiotics:Sorting the evidence from the myths[J].Medical Journal of Australia,2008,188:304-308.
[6]Aguirre M,Collins M D.Lactic acid bacteria and human clinical infection[J].Journal of Applied Microbiology,1993,75(2):95-107.
[7]张莉力,许云贺,李新华.甘薯酸浆中微生物絮凝性研究[J].食品工业科技,2010(6):172-175.
[8]张莉力,许云贺,李新华.副干酪乳杆菌副干酪亚种L1的培养条件及絮凝活性研究[J].食品工业科技,2011(3):139-145.
[9]沈雪.利用Caco-2细胞研究三种常用乳制品益生菌对肠道屏障功能影响[D].长春:吉林大学,2012.
[10]Yongtao Fei,Li Liu,Dongmei Liu.Investigation on the safety of Lactobacillus amylolyticus L6 and its fermentation properties of tofu whey[J].LWT-Food Science and Technology,2017,84:314-322.
[11]南晓芳.豆豉和腐乳中耐盐乳酸菌的筛选、鉴定及其功能特性研究[D].呼和浩特:内蒙古农业大学,2016.
[12]张灼阳,刘畅,郭晓查.乳酸茵耐药性的研究进展[J].中国微生态学杂志,2007,19(5):478-480.
[13]王梦姣,李少英,韦婉.分离自酸马奶的肠球菌属乳酸菌抗性试验[J].食品科技,2013,38(10):7-11.
[14]Shokryazdan P,Jahromi MF,Liang JB,et al.In vitro assessment of bioactivities of Lactobacillus strains as potential probiotics for humans and chickens[J].Journal of Food Science,2017,82(4):2734-2745.
[15]王梦姣,李少英,李淑芬,等.马奶及其制品中7株肠球菌属乳酸菌的安全性评价[J].食品科学,2014(17):204-208.
[16]Shekh S L,Dave J M,Vyas B R M.Characterization of Lactobacillus plantarum strains for functionality,safety and γ-amino butyric acid production[J].LWT-Food Science and Technology,2016,74:234-241.
[17]Yeo S,Lee S,Park H,et al.Development of putative probiotics as feed additives:Validation in a porcine-specific gastrointestinal tract model[J].Applied Microbiology & Biotechnology,2016,100(23):1-12.
[18]Yakabe T.Safety assessment of Lactobacillus brevis KB290 as a probiotic strain[J].Food and Chemical Toxicology,2009,49:2450-2453.
[19]Nancy J,Szabo.Safety evaluation of Lactobacillus pentosus strain b240[J].Food and Chemical Toxicology,2011,49:251-258.
[20]Tmr DA,Garcia EF,De OAA,et al.In vitro characterization of Lactobacillus strains isolated from fruit processing by-products as potential probiotics[J].Probiotics & Antimicrobial Proteins,2017:1-13.
[21]Lavillalerma L,Pérezpulido R,Martínezbueno M,et al.Characterization of functional,safety,and gut survival related characteristics of Lactobacillus strains isolated from farmhouse goat’s milk cheeses[J].International Journal of Food Microbiology,2013:136-145.
[22]Garcia Estefania F,Luciano Winnie A,Xavier Danilo E,et al.Identification of lactic acid bacteria in fruit pulp processing byproducts and potential probiotic properties of selected Lactobacillus strains[J].Frontiers in Microbiology,2016,7:1371.
[23]Runjun Dowarah,Formal analysis,Investigation.Methodology selection and characterization of probiotic lactic acid bacteria and its impact on growth,nutrient digestibility,health and antioxidant status in weaned piglets[J].Plos One,2018,13(3):e0192978.
[24]Monique S do Carmo,Francisca MF Noronha,Mariana O Arruda.Lactobacillus fermentum ATCC 23271 displays in vitro inhibitory activities against candida spp[J].Frontiers in Microbiology,2016,7:1722.
[25]李姗姗.潜在益生乳杆菌的抗生素敏感性研究[D].保定:河北农业大学,2012.
[26]Clementi F,Aquilanti L.Recent investigations and updated criteria for the assessment of antibiotic resistance in food lactic acid bacteria[J].Anaerobe,2011,17(6):394-398.
[27]周蕊,白瑶,李凤琴.食品工业用益生菌安全性评价研究进展[J].卫生研究,2012,41(3):511-514.
[28]顾旭峰,艾连中,宋馨,等.两株干酪乳杆菌的安全性评价[J].食品与发酵科技,2015,51(2):63-65.
[29]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,1999.
[30]Sanders M E,Walker D C,Walker K M,et al.Performance of commercial cultures in fluid milk applications[J].Journal of Dairy Science,1996,79:943-955.
[31]任大勇.益生乳酸杆菌的黏附及免疫调节作用研究[D].长春:吉林大学,2013.
[32]张俊娟.具有黏附性乳杆菌的筛选与特性研究[D].保定:河北农业大学,2011.
[33]Pablo F,Perez,Minnaard Y,et al.Surface propertiesof bifidobacteria strains of human origin[J].Applied and Environmental Microbiology,1998,64(1):21-26.
[34]王娉婷.传统发酵食品来源的功能性益生菌筛选及其在干酪加工中的应用[D].长春:吉林农业大学,2015.
[35]陈臣.植物乳杆菌ST-Ⅲ对肠上皮细胞的黏附性质及机理的研究[D].无锡:江南大学,2008.
[36]丁素娟,王薇薇,李爱科,等.益生菌肠道黏附性研究进展[J].饲料工业,2016,23(13):55-61.
[37]张莉.植物乳杆菌的黏附特性研究及其在益生菌干酪中的应用[D].长春:吉林大学,2013.