鸡用抗大肠杆菌感染的混合益生菌制剂的制备与评价

doi:10.16431/j.cnki.1671-7236.2020.11.031

摘要/Abstract

摘要： 本研究旨在探索益生菌对雏鸡抵抗大肠杆菌感染的影响，为微生态制剂的开发提供依据。通过牛津杯试验和细菌黏附抑制试验，初步得到体外抑制大肠杆菌效果良好的3种益生菌及它们的等比例混合菌液，分别为植物乳杆菌ZN-3、鼠李糖乳杆菌QC、丁酸梭菌HYCB和混合菌制剂X。连续30 d向7日龄SPF雏鸡分别灌服各益生菌及它们的混合制剂，并在灌服的第26~30天每天感染1次大肠杆菌XT-13，记录雏鸡存活率和体重变化。利用相同的灌服与感染方案，分析混合益生菌组雏鸡感染XT-13后的心脏、肝脏以及大肠的剖检与病理变化，并通过免疫组织化学法检测肝脏和肠道的大肠杆菌载菌量。另外，为了分析灌服混合益生菌制剂对大肠杆菌感染后雏鸡细胞因子的影响，采用非致死剂量的XT-13感染雏鸡，分别在感染后第4、8和14天检测血清中白细胞介素-10（IL-10）、白细胞介素-17（IL-17）和肝脏中禽防御素-β1（AvBD1）的水平，并测试感染后14 d血清的杀菌能力。最后，评估混合益生菌制剂对O78血清型大肠杆菌AV006感染雏鸡的广谱保护力。结果表明，各益生菌发酵液上清的抑菌效果明显，益生菌混合后可降低XT-13对DF-1细胞的黏附率（P<0.05）；灌服混合益生菌制剂对雏鸡的增重效果优于单一益生菌，且对雏鸡感染XT-13后具有较好的保护效果，存活率达到87.50%；混合益生菌制剂可显著降低雏鸡肝脏和大肠的XT-13载菌量，减轻病理损伤，调节机体血清IL-10和IL-17以及肝脏中AvBD1的水平，并提高血清的杀菌能力；大肠杆菌AV006的保护性试验结果表明，混合益生菌制剂还可降低雏鸡感染O78型大肠杆菌的死亡率。综上，植物乳杆菌ZN-3、鼠李糖乳杆菌QC和丁酸梭菌HYCB的混合制剂可促进雏鸡增重、减少载菌量、降低组织病变程度并调节宿主的免疫反应，对禽大肠杆菌病有良好的防治作用，具有开发成为抗禽大肠杆菌病益生菌制剂的巨大潜力。

关键词: 益生菌; 禽大肠杆菌病; 病理; 广谱保护

Abstract: The purpose of this study was to explore the effects of probiotics on chickens' resistance to Escherichia coli infections,and to provide a basis for the development of microecological agents.Based on the Oxford Cup test and the bacterial adhesion inhibition test,three kinds of probiotics and their equal proportion mixed bacterial liquid with good effect of inhibiting Escherichia coli in vitro were obtained initially:Lactobacillus plantarum ZN-3,Lactobacillus rhamnosus QC,Clostridium butyricum HYCB and mixed bacteria preparation X.The 7-day-old SPF chicks were continuously orally fed with probiotics for 30 days and infected with Escherichia coli XT-13 once a day from the 26th to 30th day,and the survival rate and weight change were recorded.Through the same oral and infection protocols,the chicks were necropsied after infection with XT-13,and the results of necropsy and pathology were analyzed,and the amount of bacteria in the liver and intestines was detected by immunohistochemistry.In addition,in order to analyze the effect of oral administration of mixed probiotic preparations on cytokines in chickens infected with Escherichia coli,non-lethal doses of XT-13 were selected to infect chickens,and the levels of interleukin-10 (IL-10),interleukin-17 (IL-17) in serum and avian defensin-β1 (AvBD1) in the liver were measured on the 4th,8th and 14th days after infection.At the same time,the bactericidal ability of the serum was tested on the 14th day after infection.Finally,the broad-spectrum protection ability of probiotics to O78 serotype Escherichia coli AV006 infected chickens was evaluated.The results showed that the antibacterial effect of the supernatant of each probiotic fermentation broth was obvious,and the mixing of probiotics could reduce the adhesion rate of XT-13 to DF-1 cells (P<0.05).The weight gain effect of mixed probiotics was better than that of single probiotics,which had a good protective effect on chickens infected with XT-13.The survival rate was 87.50%.The mixed probiotics significantly reduced the load of XT-13 in liver and large intestine,alleviated the pathological damage,and regulated the levels of IL-10 and IL-17 in serum and the levels of AvBD1 in the liver,while improving the bactericidal ability of the serum.In addition,the broad-spectrum protection test showed that the mixed probiotics reduced the mortality of chickens infected with O78 serotype Escherichia coli.Therefore,the mixed preparation of Lactobacillus plantarum ZN-3,Lactobacillus rhamnosus QC and Clostridium butyricum HYCB could promote chick weight gain,reduce bacterial load,reduce tissue lesions,regulate the host's immune response,and had a good prevention effect on avian colibacillosis,which had a great potential to develop into a probiotic preparation against avian colibacillosis.

Key words: probiotics; avian colibacillosis; pathology; broad spectrum protection

中图分类号:

S859

易海鸣, 张强, 赵雪锦, 杨丽, 金梅林. 鸡用抗大肠杆菌感染的混合益生菌制剂的制备与评价[J]. 中国畜牧兽医, 2020, 47(11): 3676-3686.

YI Haiming, ZHANG Qiang, ZHAO Xuejin, YANG Li, JIN Meilin. Preparation and Evaluation of Mixed Probiotics for Chicken Against Escherichia coli Infection[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(11): 3676-3686.

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