体外发酵制备生长猪结肠微生物组培养基的优化  

Optimization of Culture Medium for Pig Colonic Microbiome Prepared by Fermentation in Vitro

作　　者：白宇 颜桥 李天天[1] 周行健 何贝贝[1] 陶诗煜 张帅 王军军[1] BAI Yu;YAN Qiao;LI Tiantian;ZHOU Xingjian;HE Beibei;TAO Shiyu;ZHANG Shuai;WANG Junjun(State Key Laboratory of Animal Nutrition,College of Animal Science and Technology,China Agricultural University,Beijing 100193,China)

机构地区：[1]中国农业大学动物科技学院动物营养学国家重点实验室

出　　处：《动物营养学报》2019年第8期3719-3726,共8页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金重点项目(31630074)

摘　　要：结肠微生物组对动物后肠道的健康及其营养物质利用具有重要意义,而体内研究肠道微生物功能与机制存在诸多不便。为此,本试验在比较分析生长猪结肠食糜和食品发酵素培养基(VL培养基)营养组成的基础上,通过改变和优化VL培养基组成使其接近结肠食糜,利用体外单向连续发酵系统培养模拟微生物组,并计算模拟微生物组与结肠微生物组菌群结构相似度。结果表明:结肠食糜在粗蛋白质、可溶性糖含量方面与常用的VL培养基存在显著差异(3.09%vs.1.49%,0.43%vs.0.31%)(P<0.05);在VL培养基的基础上将粗蛋白质、可溶性糖含量调整至结肠食糜水平并分别添加糖蜜(VLMOL培养基)、玉米浆干粉(VLCSL培养基)、消化道分泌的黏蛋白(VLM培养基)及上述3种物质的组合物(ZH培养基)后发现,利用VLMOL、VLCSL、VLM和ZH培养基得到的模拟微生物组与结肠微生物组菌群结构相似度分别为68.5%、52.3%、10.7%和5.0%;在发酵特性方面,VLM OL培养基所得发酵液中短链脂肪酸总量与结肠食糜相近(21.75 mmol/L vs.26.03 mmol/L)(P>0.05),VLCSL、VLM和ZH培养基所得发酵液短链脂肪酸总量与结肠食糜相比则有显著差异(43.95、39.05和42.68 mmol/L vs.26.03 mmol/L)(P<0.05)。综上可知,根据结肠食糜优化VL培养基的粗蛋白质、可溶性糖含量,并在优化后的VL培养基中添加糖蜜所制备的VLMOL培养基是一种通过体外发酵制备生长猪结肠微生物组的可靠性较高的培养基。The colonic microbial group of animals has important significance for the health of hindgut and nutrient utilization.However,there are many inconveniences in the study of intestinal microbial function and mechanism in vivo.Therefore,this study analyzed the nutrient composition of growing pig colonic digesta and VL culture medium,and then changed and optimized the nutritional composition of the VL culture medium to consistent with colonic digesta.simulated microbiome.similar to the colonic microbiome was cultured by using the one-way continuous fermentation system in vitro,and the microbiota structural similarity between simulated microbiome and colonic microbiome was calculated.The results showed that the contents of crude protein and soluble sugar in colonic digesta was significantly different from those in VL culture medium(3.09%vs.1.49%,0.43%vs.0.31%).In addition,after adjusting the crude protein and soluble sugar contents in VL culture medium to the levels of colonic digesta,and adding the additions of molasses(VLMOL),corn syrup dry powder(VLCSL),digestive tract secreted mucin(VLM)and their combination into the culture medium,the microbiota structural similarity between the colonic microbiome and the simulated microbiome cultured by VLMOL,VLCSL,VLM and ZH culture mediums was 68.5%,52.3%,10.7%and 5.0%,respectively.In term of total short chain fatty acid concentration,the total amount of volatile fatty acids produced by VLMOL cultured colonic microbial group was more similar with that of the colonic digesta(21.75 mmol/L),whereas that in fermentation broths produced by VLCSL,VLM and ZH culture mediums were significantly different compared with colonic digesta(43.95,39.05,42.68 mmol/L vs.26.03 mmol/L).In conclusion,a highly reliable culture medium which optimized by adjusting the contents of crude protein and soluble sugar in VL culture medium based of these in colonic digesta and adding molasses into the modified VL culture medium,can be used to prepare the colonic microbiome of growing pig by fermentation in vi

关 键 词：猪 结肠 微生物组 糖蜜 体外发酵 

分 类 号：S828[农业科学—畜牧学]