猪肠道内源乳杆菌的筛选鉴定及其在液态发酵饲料中的应用  

Screening and identification of intestinal endogenous Lactobacillus in pigs and its application in liquid fermented feeds

作　　者：李耿辉 王雨君 郑太龙 王松波 王丽娜 朱灿俊 吴睿帆 江青艳 束刚 LI Genghui;WANG Yujun;ZHENG Tailong;WANG Songbo;WANG Lina;ZHU Canjun;WU Ruifan;JIANG Qingyan;SHU Gang(College of Animal Science,Guangdong Provincial Key Laboratory of Animal Nutrition Regulation,South China Animal Nutrition and Feed Science Observation Experimental Station of Ministry of Agriculture,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学动物科学学院/农业部华南动物营养与饲料科学观测实验站/广东省动物营养调控重点实验室,广东广州510642

出　　处：《江西农业大学学报》2025年第1期152-166,共15页Acta Agriculturae Universitatis Jiangxiensis

基　　金：“十四五”国家重点研发计划重点专项(2022YFD1300400)。

摘　　要：【目的】筛选适用于猪液态发酵的肠道益生菌复合菌种,探究复合菌种制备的液态发酵饲料在育肥猪生产中的饲喂效果,为后续液态发酵猪饲料的应用提供研究基础。【方法】选取28日龄、体质量为8 kg左右的健康仔公猪肠道食糜,筛选出具备产酶和产酸特性的优势益生菌,对其进行分子生物学鉴定、安全性评价,并对其中一株益生菌进行全基因组测序与功能注释;根据优势益生菌制备的不同液态发酵饲料配方的发酵效果,采用一种最佳液态发酵方案用于饲养试验;选取120头120日龄、体质量约81 kg的丹系长白猪,随机分为对颗粒料照组(CON)、液态发酵料发酵组(FLF)两组,饲养28 d,统计育肥猪采食量、体增重、料重比等生长性能指标,并对粪便进行16S rRNA测序分析其微生物组成变化。【结果】(1)根据饲料底物降解酶活筛选出了活性最高的7株单克隆菌株:Hi04(谷糠乳杆菌,Lentilactobacillus farraginis)、Hi07(谷糠乳杆菌,Lentilactobacillus farraginis)、Hp06(鼠李糖乳杆菌,Lacticaseibacillus rhamnosus)、Hp09(鼠李糖乳杆菌,Lacticaseibacillus rhamnosus)、Hd02(希氏乳杆菌,Lactobacillus hilgardii)、Lm01(嘉义乳杆菌,Lacticaseibacillus chiayiensis)和Tc04(鼠李糖乳杆菌,Lacticaseibacillus rhamnosus),7株单克隆菌对抗生素耐药性均低于商业植物乳杆菌,其中,1株嘉义乳杆菌Lm01基因组中编码的碳水化合物代谢相关的功能基因丰度非常高,表明其具有非常强的代谢潜力,并其基因组中不含毒力基因;(2)以7株复合益生菌制备的液态发酵饲料,在添加10%起始菌种为1×107 CFU/mL发酵料二次发酵4 h后,饲料pH4.2,乳酸产量201.14 mmol/L,且跟商业乳杆菌相比,对饲料蛋白的降解能力更强,降解速度更快(P<0.05);(3)饲养试验结果表明:与对照组相比,液态发酵饲料组显著提高了育肥猪平均日增重,降低了耗料增重比(P<0.05);(4)16S rRNA测序结果发现:液态发酵饲料组[Objective]This study aims to screen intestinal probiotics suitable for liquid fermentation in pigs and to determine the feeding effect of liquid-fermented feed prepared by the composite bacteria in the production of fattening pigs,thus providing a basis for the subsequent application of liquid fermented pig feed.[Method]Intestinal coeliacs were selected from healthy male piglets aged 28 days weighing approximately 8kg.The beneficial probiotics with enzyme-producing and acid-producing properties were screened,molecular biological identification and safety evaluation were conducted.and the whole-genome sequencing and functional annotation on one of the probiotic strains were performed.An optimal liquid fermentation protocol was used for feeding trials based on the fermentation effect of different liquid-fermented feed formulations produced by dominant probiotics.120 Danish Landrace pigs,120-day-old and weighing approximately 81 kg,were randomly assigned to the control group(CON,pellet feed)and the fermentation group(FLF,liquid fermentation feed)for a feeding period of 28 days.The growth performance indexes such as feed intake,body weight gain and feed/gain ratio of grow-finishing pigs were measured,and the changes of microbial composition in feces were analyzed by 16S rRNA sequencing.[Result](1)Based on the activity of feed substrate degrading enzymes,7 monoclonal strains,Hi04(Lentilactobacillus farraginis),Hi07(Lentilactobacillus farraginis),Hp06(Lacticaseibacillus rhamnosus),Hp09(Lacticaseibacillus rhamnosus),Hd02(Lactobacillus hilgardii),Lm01(Lacticaseibacillus chiayiensis) and Tc04(Lacticaseibacillus rhamnosus) with high activity were screened.All 7 monoclonal strains had lower antibiotic resistance than commercial plant Lactobacilli.The abundance of functional genes related to carbohydrate metabolism encoded in the genome of a strain of Lacticaseibacillus chiayiensis Lm01 was very high,indicating its strong metabolic potential and the absence of virulence genes in its genome.(2)The liquid fermented feed prepa

关 键 词：益生菌 液态发酵饲料 乳杆菌 育肥猪 生长性能 

分 类 号：S852.66[农业科学—基础兽医学] S815.8[农业科学—兽医学]