猪源复合乳酸菌的筛选及其改善仔猪肠炎的研究  

Screening of porcine compound lactic acid bacteria and study on their improving piglet enteritis

作　　者：刘春汝 谢霜 贾俊鹏 庾庆华 钟翔[3] 杨倩[1] LIU Chunru;XIE Shuang;JIA Junpeng;YU Qinghua;ZHONG Xiang;YANG Qian(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;Diprobio(Shanghai)Co.,Ltd.,Laboratory of Microbiology,Immunology and Metabolism,Shanghai 200335,China;College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物医学院,江苏南京210095 [2]迪辅乐生物(上海)有限公司微生物免疫代谢实验室,上海200335 [3]南京农业大学动物科技学院,江苏南京210095

出　　处：《畜牧与兽医》2023年第5期42-49,共8页Animal Husbandry & Veterinary Medicine

基　　金：国家重点研发计划专项(2018YFE0127300);国家自然科学基金(31972631)。

摘　　要：为了探究猪源复合乳酸菌对鼠伤寒沙门菌感染仔猪引起的肠道损伤的保护效果,对实验室前期从猪粪便中分离并保存的23株猪源乳酸菌进行体外耐酸、耐胆盐及体外抑菌试验筛选,并将筛选得到的乳酸菌等比例混合,形成复合乳酸菌。选取16只日龄、体重相近的断奶仔猪,随机分成4组,分别饲喂复合乳酸菌、沙门菌、复合乳酸菌+沙门菌和无菌PBS(对照组),每日记录仔猪体重变化,于6 d后屠宰采样,分别采集回肠、结肠及回肠内容物,利用苏木精-伊红(HE)染色观察回肠、结肠病理变化,过碘酸-雪夫(PAS)染色检测回肠中杯状细胞数量,16S rRNA测序检测回肠内容物中微生物组成,并测定沙门菌载量和肠道炎症相关细胞因子、黏蛋白(muc2)表达量。结果:经体外试验筛选得到5株耐酸、耐胆盐及体外抑菌效果良好的乳酸菌,等比混合组成的复合乳酸菌能有效改善鼠伤寒沙门菌导致的仔猪体重增长减缓(P=0.087),并能修复肠道组织学损伤,显著降低回肠中鼠伤寒沙门菌载量(P<0.05),抑制促炎因子白介素1β(IL-1β)、肿瘤坏死因子(TNF-α)的表达(P=0.083,P=0.073)。本研究还发现,复合乳酸菌能够显著改善沙门菌感染引起的杯状细胞的丢失并上调黏蛋白基因(muc2)表达水平(P<0.05)。此外,复合乳酸菌能够改善感染引起的仔猪肠道菌群紊乱,显著增加感染仔猪肠道中乳酸菌属的相对丰度(P<0.05),降低埃希菌属的相对丰度(P<0.05)。总之,复合乳酸菌的饲喂可减轻断奶仔猪肠道炎症,增强肠道黏液屏障,维护肠道菌群平衡,保障仔猪的肠道健康。In order to explore the protective effect of porcine compound lactic acid bacteria preparation on intestinal injury caused by Salmonella typhimurium infection in piglets,23 strains of porcine lactic acid bacteria stored in the laboratory were screened for in vitro acid resistance,bile salt resistance and in vitro bacteriostatic test,and the selected lactic acid bacteria were mixed in equal proportion to form a composite lactic acid bacteria preparation for subsequent tests.A total of 16 weaned piglets of similar age and body weight were chosen and randomly divided into 4 groups,which were fed with complex lactic acid bacteria,Salmonella,complex lactic acid bacteria+Salmonella,and sterile PBS(the control group),respectively.Body weight changes in the piglets were recorded daily,and samples were collected from them 6 days later.Samples of ileum,colon and ileum contents were collected.The ileum sample was used for hematoxylin-eosin(HE)staining to observe the pathological changes of ileum,and the number of goblet cells in the intestine of the piglets was detected by periodic acidschiff(PAS)staining.In addition,the sample was also used for the measurement of Salmonella load and the expression of intestinal inflammatory cytokines and muc2.Finally,samples of ileal contents were used for microbial 16S rRNA sequencing.The results showed that five lactic acid bacteria with good acid resistance,bile salt resistance and antibacterial effect were screened by the in vitro test.The compound lactic acid bacteria of equal ratio effectively improved the weight gain of the piglets induced by Salmonella typhimurium(P=0.087),repaired intestinal histological injury in them,significantly reduced the load of Salmonella typhimurium in ileum(P<0.05),and inhibited the expression of pro-inflammatory cytokines interleukin-1-β(IL-1β)and the tumor necrosis factor(TNF-α)(P=0.083,P=0.073).This study also found that compound lactic acid bacteria significantly improved the loss of goblet cells and up-regulated the expression of muc2 caused by S

关 键 词：断奶仔猪 复合乳酸菌 鼠伤寒沙门菌 肠道菌群 肠道健康 

分 类 号：S852.61[农业科学—基础兽医学]