【作者】 曹希亮；

【导师】 倪学勤； 曾东；

【作者基本信息】 四川农业大学 ， 预防兽医学， 2007， 硕士

【摘要】 本文通过刚出生仔猪的饲养试验，测定枯草芽孢杆菌和约氏乳杆菌对仔猪肠道黏膜免疫、T细胞免疫、肠道菌群、日增重和腹泻指数的影响。采用双抗夹心ELISA方法，测定两种微生态制剂对仔猪肠道黏膜免疫的影响，比较不同处理组粪样中的SIgA水平。结果不同处理组仔猪的SIgA水平没有显著差异，都随日龄增加而增加，5周龄达高峰，然后SIgA水平呈下降趋势。仔猪前腔静脉采血，使用流式细胞仪检测微生态制剂对仔猪T细胞亚群的影响，通过测定CD3+CD4+、CD3+CD8+、CD3+CD4+／CD3+CD8+、CD4+CD8+和CD4-CD8-的数量变化检测微生态制剂的作用效果。29日龄和35日龄时，约氏乳杆菌组CD3+CD4+和CD3+CD4+／CD3+CD8+显著高于对照组和枯草芽孢杆菌组(P＜0.05)，说明约氏乳杆菌可以提高仔猪免疫力，增强抗病能力。35日龄时，枯草芽孢杆菌组CD3+CD8+显著高于对照组和约氏乳杆菌组(P＜0.05)。其它指标与对照组差异不显著，说明枯草芽孢杆菌在增强仔猪免疫力方面没有明显作用，但是作为一种肠道外籍菌可以影响外周血CD3+CD8+的含量。仔猪外周血的CD4+CD8+数量只是35日龄时，枯草芽孢杆菌组显著高于约氏乳杆菌组，同时约氏乳杆菌组CD3+CD4+和CD3+CD4+／CD3+CD8+显著高于对照组和枯草芽孢杆菌组(P＜0.05)。三个处理组的CD4-CD8-没有显著差异，随仔猪日龄的增加有稍微下降的趋势。采集不同日龄的仔猪新鲜粪样，检测微生态制剂和对照组的厌氧菌和需氧菌的数量。35日龄时，对照组大肠杆菌显著高于约氏乳杆菌组(P＜0.05)，其他时间各个处理组差异不显著；在7日龄、14日龄和29日龄时枯草芽孢杆菌组和约氏乳杆菌组梭菌显著高于对照组(P＜0.05)；在7日龄和21日龄，约氏乳杆菌处理组消化球菌显著高于对照组(P＜0.05)；肠球菌数量只是在仔猪断奶时，约氏乳杆菌组显著高于对照组(P＜0.05)；乳酸杆菌的数量只是在7日龄的时候显著高于对照组；双歧杆菌在不同处理组没有显著差异。说明添加微生态制剂一定程度上起到了增加肠道厌氧菌的数量，断奶时肠球菌数量的增加说明断奶应激时微生态制剂起到了平衡肠道厌氧菌和需氧菌比例的作用。记录仔猪的每天腹泻情况，以检测微生态处理组在控制仔猪腹泻方面的作用效果。并于仔猪刚出生和试验结束时记录仔猪的窝重，用以计算仔猪的日增重。添加微生态制剂对仔猪日增重的影响效果显著，尤其是约氏乳杆菌末重和日增重都显著高于对照组(P＜0.05)。而微生态制剂处理组在控制腹泻方面也起到了明显的效果，腹泻指数微生态制剂处理组显著低于对照组(P＜0.05)。说明微生态制剂可以很好的控制仔猪腹泻和提高仔猪的日增重。更多还原

【Abstract】 The influence of the probiotic bacterium B.subtilis and L.johnsonii on the immunesystem, intestinal microbial population, average daily liveweight gain and diarrhea scorewere determined in a feeding experiment with piglets. Capture enzyme-linkedimmunosorbent assays were used to quantify the total concentrations of IgA in faecesof different groups. Blood of piglets was sampled by means of vena cava anterior, and flowcytometry detected influence of different probiotics on T-cell subpopulation. Quantitativechange of CD3+CD4+, CD3+CD8+, CD3+CD4+/CD3+CD8+, CD4+CD8+ andCD4-CD8- reflected effect of probiotics. Fresh faeces of different ages’piglets werecollected to detect intestinal microbial population and secretory IgA. Diarrhea condition ofpiglets was recorded to find effect of probiotics in controlling dearrhea. At the beginning.and the end of the experiment, weigh to calculate average daily liveweight gain.Total IgA of piglets in different groups appeared to be unaffected. Piglets of differentgroups showed similar SIgA levels up to 5 weeks after birth with a slight low tendency. Atan age of 35 days, percentage of CD3+CD8+ of B.subtilis group was significantlyhigher than control and L.johnsonii groups (P＜0.05). At the age of 29 days and 35 days, Asto percentage of CD3+CD4+ and proportion of CD3+CD4+/CD3+CD8+, L.johnsoniigroup was significantly higher than control and B.subtilis groups (P＜0.05), suggestingL.johnsonii can improve piglets’immunity and enhance disease resistance,and difference ofB.subtilis and control groups was not obvious. But CD3+CD8+ was obviously higher thananother two groups, suggesting B. subtilis can not enhance piglets’immune obviously, andthen change percentage of CD3+CD8+ in peripheral blood as an extrinsic bacteria. Pigletsthat was supplemented B.subtilis group displayed higher levels of CD4+CD8+ thanL.johnsonii group at the age of 35 days, just then percentage of CD3+CD4+ andCD3+CD4+/CD3+CD8+ of L.johnsonii group was obviously higher than control and B.subtilis groups, suggesting that CD4+CD8+ can not evaluate piglet’health here, at thesame time can not predicate role of CD4+CD8+. Percentage of CD4-CD8- in peripheralblood of different groups was not obviously different, and took on a slight low tendencywhen piglets got older from birth to 5 weeks.With regard to influence ofprobiotics on intestinal microbial population, at an age of 35days, E. coli of control group was obviously higher than L.johnsonii group, but differentgroups were not obvious difference in the remaining time; As to Clostridium, when pigletswere 7,14 and 29 days, B.subtilis and L.johnsonii groups were obviously higher thancontrol group (P＜0.05); At the age of piglets’7 days and 21 days, Peptococcus ofL.johnsonii group was obviously higher than control group (P＜0.05); When piglets wereweaned, Enterococcus of L.johnsonii group was obviously higher than control; At theage of 7 days, L.johnsonii of probiotics groups was obviously higher than control group;Bifidobacterium of different groups appeared to be unaffected. These analysis suggestedprobiotics can increase anaerobian in the intestinal tract to some extent. When piglets wereweaned, enterococcus of probiotics groups increased, which could suggest probiotics couldbalanced the minute ecology of intestinal tract that was proportion of anaerobian andaerobe.Influence of probiotics on piglets’ average daily liveweight gain was obvious, especiallyend-weight and day-weight of L.johnsonii group were obviously higher than controlgroup. Probiotics could effectively control piglets’ diarrhea, and diarrhea index wasobviously lower than control group (P＜0.05). In conclusion, probiotics could controldiarrhea and elevate average daily liveweight gain effectively.更多还原