甜叶菊绿原酸增强大肠杆菌感染蛋雏鸡免疫力研究  

Effects of Stevia Chlorogenic Acid on Enhancing the Immunity of E.coli O_(78)-infected Layers

作　　者：卓春柳 王方方 钟翠红[1] 王斌[1] 石玉祥[1] 徐美利 武亚南 张永英[1] ZHUO Chunliu;WANG Fangfang;ZHONG Cuihong;WANG Bin;SHI Yuxiang;XU Meili;WU Ya’nan;ZHANG Yongying(Hebei University of Engineering,Handan 056000,China;Chenguang Biotech Group Co. Ltd., Handan 056000, China)

机构地区：[1]河北工程大学,邯郸056000 [2]晨光生物科技集团股份有限公司,邯郸056000

出　　处：《畜牧兽医学报》2021年第8期2200-2212,共13页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：河北省科技厅立项项目(19046637Z);河北省植物天然色素产业研究院(20181114)。

摘　　要：旨在评价甜叶菊绿原酸增强人工腹气囊感染大肠杆菌O_(78)蛋雏鸡免疫力的作用效果,为功能性抗生素替代品研发提供基础参数支持。本试验随机将1日龄、体重无显著差异的健康海兰蛋鸡360只分为6组:空白对照组(C)、大肠杆菌O_(78)处理组(EC0)、1.0 g·L^(-1)杜仲素+大肠杆菌O_(78)处理组(ED1)、1.0 g·L^(-1)甜叶菊绿原酸+大肠杆菌O_(78)处理组(EC1)、2.0 g·L^(-1)甜叶菊绿原酸+大肠杆菌O_(78)处理组(EC2)、4.0 g·L^(-1)甜叶菊绿原酸+大肠杆菌O_(78)处理组(EC4),预饲7 d后开始正式试验。第7天时将大肠杆菌O_(78)通过腹气囊感染蛋雏鸡,饮水投喂药物,每天1次,连用3 d。随后通过ELISA法检测血清IL^(-1)β、IL-2、IL-6、IgM、IgA、TNF-α水平;RT-qPCR检测空肠和回肠IL^(-1)β、IL-2、TNF-α、Claudin-1和ZO-1基因表达;高通量测序分析盲肠内容物微生物种类。结果显示:1)腹气囊感染大肠杆菌O_(78)显著增加了蛋雏鸡死亡率(P<0.05),而甜叶菊绿原酸处理组(EC2、EC4)蛋雏鸡的死亡率显著降低(P<0.05)。2)甜叶菊绿原酸对大肠杆菌O_(78)感染蛋雏鸡血清IgA和IgM含量有提高趋势,可不同程度降低血清炎性因子含量,其中EC1、EC2、EC4组血清TNF-α含量显著降低(P<0.05);EC2显著降低大肠杆菌O_(78)感染蛋雏鸡回肠促炎细胞因子IL^(-1)β、IL-2、TNF-α的基因表达(P<0.05)。3)甜叶菊绿原酸可促进蛋雏鸡空肠紧密连接蛋白基因表达,改善腹气囊感染大肠杆菌O_(78)对肠道屏障的损伤。4)腹气囊感染大肠杆菌O_(78)导致鸡肠道特有OTUs增加,增加肠道拟杆菌门、变形菌门和梭杆菌门的相对丰度,降低厚壁菌门的相对丰度;而甜叶菊绿原酸处理组(EC2)蛋雏鸡盲肠厚壁菌门的相对丰度升高,拟杆菌门和变形菌门的相对丰度降低。甜叶菊绿原酸可增强腹气囊感染大肠杆菌O_(78)蛋雏鸡机体的免疫功能,抵御大肠杆菌O_(78)对蛋雏鸡的侵袭,其中应用剂量为2.0 g·L^(-1This study aimed to evaluate the effect of stevia chlorogenic acid on enhancing immunity of layers infected with E.coli O_(78) via abdominal air sac artificially,and to provide basic parameter support for the development of functional antibiotic substitutes.A total of 3601-day-old healthy Hailan layers with no significant difference in body weight were randomly divided into 6 groups:control group(C),E.coli O_(78) treatment group(EC0),1.0 g·L^(-1) eucommdin+E.coli O_(78) treatment group(ED1),1.0 g·L^(-1) stevia chlorogenic acid+E.coli O_(78) treatment group(EC1),2.0 g·L^(-1) stevia chlorogenic acid+E.coli O_(78) treatment group(EC2),4.0 g·L^(-1) stevia chlorogenic acid+E.coli O_(78) treatment group(EC4),the formal test started after 7 days of pre-feeding.On the 7th day,the layers was infected with E.coli O_(78)through the abdominal air sac.The treatment lasted for 3 days,and medicine was dissolved into drinking water once a day.Subsequently,the serum levels of IL^(-1)β,IL-2,IL-6,IgM,IgA and TNF-αwere detected by ELISA;the expression level of IL^(-1)β,IL-2,TNF-α,Claudin-1 and ZO-1 genes in jejunum and ileum were detected by RT-qPCR;the high-throughput sequencing was used to analyze the types of microorganisms in the cecum contents.The results showed that:1)E.coli O_(78) significantly increased the mortality of layers(P<0.05),while the mortality of layers in EC2 and EC4 groups were significantly reduced(P<0.05).2)Stevia chlorogenic acid had a tendency to increase serum IgA and IgM levels in E.coli O_(78)-infected layers,and reduce serum inflammatory factor levels to varying degrees.Among them,serum TNF-αlevels of layers in EC1,EC2 and EC4 groups were significantly reduced(P<0.05);the gene expression of ileal pro-inflammatory cytokines IL^(-1)β,IL-2 and TNF-αin layers infected with E.coli O_(78) were significantly reduced in EC2 group(P<0.05).3)Stevia chlorogenic acid could promote the expression of jejunum tight junction protein gene,and improve the intestinal barrier damage caused by E.coli O_(78) infect

关 键 词：蛋雏鸡 绿原酸 大肠杆菌O_(78) 腹气囊感染 免疫功能 

分 类 号：S831.4[农业科学—畜牧学]