嗜酸乳杆菌发酵中药对产肠毒素大肠杆菌K88所致腹泻的防治作用

doi:10.16431/j.cnki.1671-7236.2018.03.030

摘要/Abstract

摘要：

本试验旨在研究嗜酸乳杆菌发酵中药对产肠毒素大肠杆菌（enterotoxigenic Escherichia coli，ETEC）K88所致腹泻小鼠的影响。选取6~8周龄巴比西小鼠（BALB/c）100只，分为4组，分别为空白对照组（Ⅰ组）、大肠杆菌感染模型组（Ⅱ组）、中药发酵组（Ⅲ组）及中药水提组（Ⅳ组），其中Ⅱ、Ⅲ和Ⅳ组小鼠均用"抗生素+大肠杆菌"模式造模。Ⅰ、Ⅱ组饲喂不含任何药物的基础日粮，Ⅲ、Ⅳ组分别在基础日粮中添加10%中药发酵液、10%中药水提液。统计分析各组小鼠腹泻率、腹泻指数、免疫器官指数、肠道菌群及细胞因子IFN-γ、IL-4的含量。结果显示，ETEC K88感染小鼠36 h后，与Ⅱ组相比，Ⅲ、Ⅳ组小鼠腹泻指数（P < 0.05）与腹泻率降低，且Ⅲ组腹泻指数显著低于Ⅳ组（P < 0.05），Ⅲ、Ⅳ组盲肠内大肠杆菌数量较Ⅱ组降低，双歧杆菌及乳酸菌数量提高，且Ⅲ组效果更优；Ⅲ、Ⅳ组的免疫器官指数均高于Ⅰ和Ⅱ组，且与Ⅱ组相比，Ⅲ组的脾脏指数、胸腺指数及肝脏指数分别提高了48.37%、29.57%、36.88%，且差异均显著（P < 0.05）；Ⅲ、Ⅳ组IFN-γ/IL-4比值较Ⅱ组提高，且Ⅲ组高于Ⅳ组。综上所述，乳酸菌发酵中药液具有降低腹泻小鼠腹泻率及腹泻指数，提高免疫器官指数，维持肠道菌群平衡及快速消除体内炎症反应等作用，可为开发防制ETEC所致的仔猪腹泻产品提供参考。

关键词: 嗜酸乳杆菌; 发酵中药; 产肠毒素大肠杆菌; 腹泻率

Abstract:

The experiment was conducted to investigate the effects of Lactobacillus acidophilus fermented traditional Chinese medicine on diarrhea mice induced by enterotoxigenic Escherichia coli (ETEC) K88.100 6-8 weeks old BALB/c mice were chosen and divided into 4 groups:Groups Ⅰ,Ⅱ,Ⅲ and Ⅳ.Group Ⅰ was the blank control group,while the mice in groups Ⅱ,Ⅲ and Ⅳ were induced by ETEC K88.The mice in groups Ⅰ and Ⅱ were fed with basal diet without any medicine,while that in groups Ⅲ and Ⅳ were fed with basal diet supplemented with 10% traditional Chinese medicine fermentation liquor and 10% traditional Chinese medicine aqueous extract,respectively.The diarrhea rate,diarrhea index,immune organ index,intestinal flora and cytokine IFN-γ and IL-4 level were statistically analyzed.The results showed that,after 36 h of K88 infection,the diarrhea index (P < 0.05) and diarrhea rate of groups Ⅲ and Ⅳ were decreased compared with group Ⅱ,and the diarrhea index of group Ⅲ were significantly lower than group Ⅳ (P < 0.05),and the number of Escherichia coli in the cecum was decreased,while the number of Bifidobacteria and Lactobacillus bacteria were increased.The immune indexes of groups Ⅲ and Ⅳ were higher than that of groups Ⅰ and Ⅱ.Compared with group Ⅱ,the spleen index,thymus index and liver index of group Ⅲ were increased by 48.37%,29.57% and 36.88%,respectively (P < 0.05).Compared with group Ⅱ,the IFN-γ/IL-4 ratio of groups Ⅲ and Ⅳ was increased,and that of group Ⅲ was higher than group Ⅳ.In conclusion,Lactobacillus acidophilus fermented traditional Chinese medicine could reduce the diarrhea rate and diarrhea index,improve the index of immune organs,maintain the balance of intestinal flora and eliminate the inflammatory reaction in diarrhea,providing reference for the development of products for piglet diarrhea induced by ETEC.

Key words: Lactobacillus acidophilus; fermented Chinese medicine; enterotoxigenic Escherichia coli(ETEC); diarrhea rate

中图分类号:

S816.7

谷巍, 王丽荣, 孙明杰, 陈静, 陈振, 曾佳佳, 李士栋, 王春凤, 单宝龙, 徐海燕. 嗜酸乳杆菌发酵中药对产肠毒素大肠杆菌K88所致腹泻的防治作用[J]. 《中国畜牧兽医》, 2018, 45(3): 798-806.

GU Wei, WANG Lirong, SUN Mingjie, CHEN Jing, CHEN Zhen, ZENG Jiajia, LI Shidong, WANG Chunfeng, SHAN Baolong, XU Haiyan. Effect of Lactobacillus acidophilus Fermentated Traditional Chinese Medicine on Diarrhea Induced by Enterotoxigenic Escherichia coli K88[J]. , 2018, 45(3): 798-806.

参考文献

[1] 丁亚红,丁爱云,王秋娟,等.大肠埃希菌多价卵黄抗体的研制及其体外抑制作用[J].中国兽医杂志,2008,44(1):26-27. DING Y H,DING A Y,WANG Q J,et al.Preparation of egg yolk antibody from Escherichia coli and its inhibitory effect in vitro[J].Chinese Journal of Veterinary Medicine,2008,44(1):26-27.(in Chinese)
[2] FAIRBROTHER J M.Disease of Swine[M].USA:Iowa State University Press,1992.
[3] 储卫华,高国峰.卵黄抗体被动免疫防治仔猪产肠毒素大肠埃希菌性腹泻[J].中国兽医学报,2006,26(6):616-618. CHU W H,GAO G F.Egg yolk antibody passive immunization control piglets enterotoxigenic Escherichia coli diarrhea[J].Chinese Journal of Veterinary Science,2006,26(6):616-618.(in Chinese)
[4] 王智云,孙玉刚,王麟,等.乌梅的药理活性研究进展[J].实用临床医药杂志,2015,19:200-202. WANG Z Y,SUN Y G,WANG L,et al.Advances in research on pharmacological activity of Fructus mume[J].Journal of Clinical Medicine in Practice,2015,19:200-202.(in Chinese)
[5] 林云,江林,蒋健,等蒲公英的药理作用研究进展[J].中国现代中药,2011,8:42-47. LIN Y,JIANG L,JIANG J,et al.Research progress on pharmacological effects of Taraxacum mongo-licum[J].Modern Chinese Medicine,2011,8:42-47.(in Chinese)
[6] 郑兰娟,罗艳萍,汪玉娇,等.五倍子抗菌抗炎作用研究进展[J].中国病原生物学杂志,2011,11:868-869. ZHENG L J,LUO Y P,WANG Y J,et al.Research progress on antibacterial and anti-inflammatory effects of Galla chinensis[J].Journal of Pathogenic Biology,2011,11:868-869.(in Chinese)
[7] 张菊,李金敏,于艳,等.微生物发酵中草药的研究进展及在畜牧水产养殖中的应用[J].饲料广角,2013,11:36-38. ZHANG J,LI J M,YU Y,et al.Research progress of microbial fermentation of Chinese herbal medicine and its application in animal husbandry and aquacul-ture[J].Feed China,2013,11:36-38.(in Chinese)
[8] 史洪涛,乔宏兴,王永芬,等.乳酸菌发酵扶正解毒散复合制剂的制备及其在肉鸡上的应用研究[J].中国畜牧兽医,2016,43(4):1066-1071. SHI H T,QIAO H X,WANG Y F,et al.Preparation of compound preparation of Fuzheng Jiedu powder fermented by lactic acid bacteria and its application in broilers[J].China Animal Husbandry & Veterinary Medicine,2016,43(4):1066-1071.(in Chinese)
[9] 谢炎福,代春梅,杜亚.益生菌发酵复方中药对黄河鲤生长性能和免疫性能的影响[J].黑龙江畜牧兽医,2015,15:198-200. XIE Y F,DAI C M,DU Y.Effects of probiotic fermented compound Chinese medicine on growth performance and immune function of the Yellow River carp[J].Heilongjiang Animal Science and Veterinary Medicine,2015,15:198-200.(in Chinese)
[10] 林标声,吴江文,戴爱玲,等.银杏叶-桑叶中药发酵制剂防治仔猪病毒性胃肠炎的应用效果研究[J].中国兽医杂志,2016,9:56-58. LIN B S,WU J W,DAI A L,et al.Application effect of Ginkgo leaf-mulberry leaf Chinese medicine fermentation preparation on prevention and treatment of viral gastroenteritis in piglets[J].Chinese Journal of Veterinary Medicine,2016,9:56-58.(in Chinese)
[11] 王建国,李秀丽,范寰,等.乳酸菌发酵物与中药配伍对预防鸡大肠杆菌病协同作用的研究[J].中国畜牧兽医,2015,42(7):1890-1896. WANG J G,LI X L,FAN H,et al.Synergistic effect of Lactobacillus fermented matter and traditional Chinese medicine on the prevention of colibacillosis in chickens[J].China Animal Husbandry & Veterinary Medicine,2015,42(7):1890-1896.(in Chinese)
[12] 秦波,林显华,王丽荣,等.乳酸菌发酵中药制剂防制鸡球虫病条件的优化及应用[J].中国畜牧兽医,2015,42(11):3093-3098. QIN B,LIN X H,WANG L R,et al.Optimization and application of lactic acid bacteria fermented Chinese medicine preparation to prevent chicken coccidio-sis[J].China Animal Husbandry & Veterinary Medicine,2015,42(11):3093-3098.(in Chinese)
[13] 余立功.慢性腹泻辩证分型论治体会[J].中医药临床杂志,2005,17(4):340-341. YU L G.Dialectical treatment of chronic diarrhea[J].Clinical Journal of Traditional Chinese Medicine,2005,17(4):340-341.(in Chinese)
[14] 王小婷,蒋国政,王海滨,等.乌梅肉提取物抗腹泻作用研究[J].中国现代应用药学,2016,4:407-410. WANG X T,JIANG G Z,WANG H B,et al.Study on the anti-diarrhea effect of mume extract[J].Chinese Journal of Modern Applied Pharmacy,2016,4:407-410.(in Chinese)
[15] 石壁,狄莹,何有节,等.鞣质的药理活性[J].中草药,1998,23(7):487-490. SHI B,DI Y,HE Y J,et al.Pharmacological activity of tannins[J].Chinese Traditional and Herbal Drugs,1998,23(7):487-490.(in Chinese)
[16] 季宇彬.中药有效成分药理与应用[M].哈尔滨:黑龙江科学技术出版社,1995. JI Y B.Pharmacology and Application of Active Ingredients in Traditional Chinese Medicine[M].Harbin:Heilongjiang Science and Technology Press,1995.(in Chinese)
[17] 刘现兵,杜镇镇,王冬,等.五倍子的体外抑菌作用研究[J].中国实用医药,2008,7:96-97. LIU X B,DU Z Z,WANG D,et al.Study on bacteriostasis of Galla chinensis in vitro[J].Chinese Practical Medicine,2008,7:96-97.(in Chinese)
[18] 韩战强,朱金凤,蒋晓飞,等.生物发酵中药对围产期母猪繁殖性能和仔猪健康的影响[J].畜禽业,2014,8:38-39. HAN Z Q,ZHU J F,JIANG X F,et al.Effects of Chinese herbal medicine on reproductive performance and piglet health of perinatal sows[J].Livestock and Poultry Industry,2014,8:38-39.(in Chinese)
[19] 贺晓玉.蛹虫草菌固体发酵五味子药渣工艺的优化及其产物对断奶仔猪肠道形态结构和功能的影响研究[J].雅安:四川农业大学,2014. HE X Y.Optimization of solid state fermentation process of Cordyceps chinensis dregs by Cordyceps militaris and effects of its products on intestinal morphology,structure and function of weaned piglets[D].Ya'an:Sichuan Agricultural University,2014.(in Chinese)
[20] 陈福星,陈文英,郝艳霜.蒲公英对小鼠免疫器官的影响[J].动物医学进展,2008,29(4):10-12. CHEN F X,CHEN W Y,HAO Y X.Effects of Taraxacum mongolicum on immune organs in mice[J].Progress in Veterinary Medicine,2008,29(4):10-12.(in Chinese)
[21] 于新慧,石学魁,张晓莉,等.蒲公英对小鼠免疫功能的调节研究[J].牡丹江医学院学报,2008,29(4):11-13. YU X H,SHI X K,ZHANG X L,et al.Effects of Taraxacum mongolicum on immune organs in mice[J].Journal of Mudanjiang Medical University,2008,29(4):11-13.(in Chinese)
[22] 郭剑英,颜诚,王杰豪,等.饲料中添加复方中药对小鼠血液生理生化指标和免疫器官指数的影响[J].中国畜牧兽医,2012,39(5):240-243. GUO J Y,YAN C,WANG J H,et al.Effects of compound Chinese herbal medicine on blood physiological and biochemical indexes and immune organ index in mice[J].China Animal Husbandry & Veterinary Medicine,2012,39(5):240-243.(in Chinese)
[23] 何方,王水莲,李文平.古汉养生精中药渣发酵制剂对小鼠耐力及免疫力的影响[J].中兽医医药杂志,2015,6:8-10. HE F,WANG S L,LI W P.Effects of guhanyangshengjing solution dregs fermentation preparation on endurance and immunity in mice[J].Journal of Traditional Chinese Veterinary Medicine,2015,6:8-10.(in Chinese)
[24] 张桂枝,邢启银,靳双星.复方中药混菌发酵制剂对AA肉鸡免疫功能的影响[J].中国兽医杂志,2016,7:52-55. ZHANG G Z,XING Q Y,JIN S X.Effect of compound Chinese herbal medicine mixed fermentation preparation on immune function of AA broilers[J].Chinese Journal of Veterinary Medicine,2016,7:52-55.(in Chinese)
[25] 黄福佳.WB800N-CC31工程菌发酵产物对大鼠生长、消化、肠道菌群及免疫的影响[D].大庆:黑龙江八一农垦大学,2016. HUANG F J.Effects of WB800N-CC31 fermentation products on growth,digestion,intestinal flora and immunity in rats[D].Daqing:Heilongjiang Bayi Agricultural University,2016.(in Chinese)
[26] 韩春杨,刘翠艳,牛钟相.中药制剂发酵前后成分的变化及对肉鸡部分免疫指标和生长的影响[J].畜牧兽医学报,2005,36(11):1123-1127. HAN C Y,LIU C Y,NIU Z X.Changes of components before and after fermentation and influence on some immune indexes and growth of broilers[J].Acta Veterinaria et Zootechnica Sinica,2005,36(11):1123-1127.(in Chinese)
[27] 俞晓辉,姚文,施学仕.大豆发酵蛋白替代鱼粉对断奶仔猪生产性能和肠道主要菌群的影响[J].动物营养学报,2008,21(1):46-51. YU X H,YAO W,SHI X S.Effects of soybean protein replacement on fish meal on production performance and intestinal microflora in weaned piglets[J].Chinese Journal of Animal Nutrition,2008,21(1):46-51.(in Chinese)
[28] DUIN M L.The immunological synapse:Signaling networks coordinating T cell migration and antigen recognition[J].Molecular Biology of the Cell,2001,12:394.
[29] BRANDTZAEG P.Homeoatic impact of indigenous microbiota and secretory immunity[J].Beneficial Microbes,2010,1(3):211-227.
[30] GHADIMI D,FOLER R,DE VRESE M,et al.Effects of probiotic bacteria and their genomic DNA on TH1/TH2-cytokine production by peripheral blood mononuclear cells(PBMCs) of healthy and allergic subjects[J].Immunobiology,2008,213(8):677-692.
[31] 杨帆.健脾补肾止泻汤治疗慢性腹泻50例[J].陕西中医学院学报,2010,33(2):27-28. YANG F.Treatment of 50 cases of chronic diarrhea with Jianpi Bushen Zhixie Decoction[J].Journal of Shaanxi College of Traditional Chinese Medicine,2010,33(2):27-28.(in Chinese)