表乳糖对长双歧杆菌抑制产气荚膜梭菌生长的影响
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摘要
产气荚膜梭菌是一种常见的人畜禽共患的病原菌,可引起人的急性食物中毒和慢性肠道感染,或导致动物的坏死性肠炎等,严重危害人体健康和畜禽养殖业的发展。采用对长双歧杆菌和产气荚膜梭菌单独培养和共培养的方式,研究表乳糖及其同分异构体乳糖、乳果糖等特异性碳源对双歧杆菌的生长特性及其抑制产气荚膜梭菌生长的影响。结果表明,长双歧杆菌能够在以表乳糖为碳源的m TPY培养基中生长良好,并显现出与乳糖相似的生长趋势。在与产气荚膜梭菌共培养的过程中,长双歧杆菌能够显著地抑制产气荚膜梭菌的生长,且表乳糖相较于乳糖和乳果糖,能显著提高长双岐杆菌的存活率。还利用牛津杯法对长双歧杆菌培养上清液对产气荚膜梭菌的抑菌效果进行评价,证明其培养上清液对于产气荚膜梭菌具有较好的抑菌活性。上述结果为应用长双歧杆菌抑制产气荚膜梭菌提供理论依据,对深入研究表乳糖调控机制具有重要的指导意义。
Clostridium perfringens is a common pathogen in human,livestock and poultry,which can cause acute food poisoning,chronic intestinal infection in human or necrotizing enteritis in animals. It seriously endangers human health and the development of livestock and poultry breeding. In this paper,the effects of the specific carbon sources such as epilactose and its isomers lactose and lactulose were studied by the single-strain culture of Bifidobacterium longum and the co-culture of B. longum and C. perfringens. The results showed that B. longum could grow well in m TPY medium with epilactose as a carbon source and showed a similar growth trend as lactose. In addition,B.longum could significantly inhibit the growth of C. perfringens in a co-culture system,and the viability of B. longum in the epilactose medium was significantly better than that in lactose and lactulose ones. Moreover,an oxford cup test was also applied to measure the antibacterial effect of B. longum on C. perfringens,showing that the culture supernatant of B. longum had a good antibacterial activity against C. perfringens. These results provided theoretical reference for the inhibition of C. perfringens by B. longum,and also provide important guidance for further study of epilactose and its regulation mechanism.
Clostridium perfringens is a common pathogen in human,livestock and poultry,which can cause acute food poisoning,chronic intestinal infection in human or necrotizing enteritis in animals. It seriously endangers human health and the development of livestock and poultry breeding. In this paper,the effects of the specific carbon sources such as epilactose and its isomers lactose and lactulose were studied by the single-strain culture of Bifidobacterium longum and the co-culture of B. longum and C. perfringens. The results showed that B. longum could grow well in m TPY medium with epilactose as a carbon source and showed a similar growth trend as lactose. In addition,B.longum could significantly inhibit the growth of C. perfringens in a co-culture system,and the viability of B. longum in the epilactose medium was significantly better than that in lactose and lactulose ones. Moreover,an oxford cup test was also applied to measure the antibacterial effect of B. longum on C. perfringens,showing that the culture supernatant of B. longum had a good antibacterial activity against C. perfringens. These results provided theoretical reference for the inhibition of C. perfringens by B. longum,and also provide important guidance for further study of epilactose and its regulation mechanism.
引文
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[12]文钰,杨汝德.猪源双歧杆菌的分离纯化及初步鉴定[J].现代食品科技,2007,23(4):11-13.Wen Y,Yang R D. Isolation,purification and identification ofBifidobacterium pullorum from piglets feces[J]. Modern FoodSci. Technol.,2007,23(4):11-13.
[13]赵胜娟,罗红霞,杨海莺,等.双歧杆菌Bbmn01对小鼠肠道菌群的影响[J].食品科学,2008,29(2):394-397.Zhao S J, Luo H X, Yang H Y, et al.. Effect ofBifidobacterium BBMN01 on mouse intestinal flora[J]. FoodSci.,2008,29(2):394-397.
[14] Krewinkel M,Gosch M, Rentschler E, et al.. Epilactoseproduction by 2 cellobiose 2-epimerases in natural milk[J]. J.Dairy Sci.,2014,97(1):155-161.
[15] Saburi W, Yamamoto T, Taguchi H, et al.. Practicalpreparation of epilactose produced with cellobiose 2-epimerasefrom Ruminococcus albus NE1[J]. Biosci. Biotechnol.Biochem.,2010,74(8):1736-1737.
[16] Chen Q,He W,Yan X,et al.. Construction of an enzymaticroute using a food-grade recombinant Bacillus subtilis for theproduction and purification of epilactose from lactose[J]. J.Dairy Sci.,2018,101(3):1872-1882.
[17] Ouigley E M M. The microbiota in gastrointestinal pathophy-siology[M]. Amsterdam:Elsevier,2017:139-141.
[18] Turroni F, van Sinderen D, Ventura M. Genomics andecological overview of the genus Bifidobacterium[J]. Int. J.Food Microbiol.,2011,149(1):37-44.
[19] Gibson G R,Wang X. Regulatory effects of Bifidobacteria onthe growth of other colonic bacteria[J]. J. Appl. Bacteriol.,1994,77(4):412-420.
[2] Uzal F A,Freedman J C,Shrestha A,et al.. Towards anunderstanding of the role of Clostridium perfringens toxins inhuman and animal disease[J]. Future Microbiol.,2014,9(3):361-377.
[3]陈小云,关孚时,张存帅,等.产气荚膜梭菌主要外毒素最新研究进展[J].中国兽药杂志,2005,39(6):29-33.
[4] Li J,Uzal F,Mc Clane B. Clostridium perfringens sialidases:Potential contributors to intestinal pathogenesis and therapeutictargets[J]. Toxins,2016,8(11):1-15.
[5] Ahn J B,Hwang J K,Kim C T,et al.. Bifidogenic effect ofglucooligosaccharide prepared from glucose by extrusion process[J]. J. Microbiol. Biotechnol.,1997,7(3):174-179.
[6] Lee J C,Na K,Yun J M,et al.. In vitro biridogenic effect ofnondigestible oligosaccharides isolated from red ginseng marc[J]. J. Microbiol. Biotechnol.,2001,11(5):858-862.
[7] Biavati B,Mattarelli P. The family Bifidobacteriaceae[J].Prokaryotes,2006(3):322-382.
[8] Tacconi S,Sgorbati B,Modesto M,et al.. Carbohydrate stress-related response in Bifidobacterium pseudolongum subsp.globosum[J]. Ann. Microbiol.,2012,62(4):1751-1756.
[9] Ito S,Taguchi H,Hamada S,et al.. Enzymatic properties ofcellobiose 2-epimerase from Ruminococcus albus and thesynthesis of rare oligosaccharides by the enzyme[J]. Appl.Microbiol. Biotechnol.,2008,79(3):433-441.
[10] Mu W,Li Q,Fan C,et al.. Recent advances on physiologicalfunctions and biotechnological production of epilactose[J].Appl. Microbiol. Biotechnol.,2013,97(5):1821-1827.
[11] Watanabe J,Nishimukai M, Taguchi H, et al.. Prebioticproperties of epilactose[J]. J. Dairy Sci.,2008,91(12):4518-4526.
[12]文钰,杨汝德.猪源双歧杆菌的分离纯化及初步鉴定[J].现代食品科技,2007,23(4):11-13.Wen Y,Yang R D. Isolation,purification and identification ofBifidobacterium pullorum from piglets feces[J]. Modern FoodSci. Technol.,2007,23(4):11-13.
[13]赵胜娟,罗红霞,杨海莺,等.双歧杆菌Bbmn01对小鼠肠道菌群的影响[J].食品科学,2008,29(2):394-397.Zhao S J, Luo H X, Yang H Y, et al.. Effect ofBifidobacterium BBMN01 on mouse intestinal flora[J]. FoodSci.,2008,29(2):394-397.
[14] Krewinkel M,Gosch M, Rentschler E, et al.. Epilactoseproduction by 2 cellobiose 2-epimerases in natural milk[J]. J.Dairy Sci.,2014,97(1):155-161.
[15] Saburi W, Yamamoto T, Taguchi H, et al.. Practicalpreparation of epilactose produced with cellobiose 2-epimerasefrom Ruminococcus albus NE1[J]. Biosci. Biotechnol.Biochem.,2010,74(8):1736-1737.
[16] Chen Q,He W,Yan X,et al.. Construction of an enzymaticroute using a food-grade recombinant Bacillus subtilis for theproduction and purification of epilactose from lactose[J]. J.Dairy Sci.,2018,101(3):1872-1882.
[17] Ouigley E M M. The microbiota in gastrointestinal pathophy-siology[M]. Amsterdam:Elsevier,2017:139-141.
[18] Turroni F, van Sinderen D, Ventura M. Genomics andecological overview of the genus Bifidobacterium[J]. Int. J.Food Microbiol.,2011,149(1):37-44.
[19] Gibson G R,Wang X. Regulatory effects of Bifidobacteria onthe growth of other colonic bacteria[J]. J. Appl. Bacteriol.,1994,77(4):412-420.