饲粮添加溶菌酶对断奶仔猪生长性能、肠道形态、免疫功能和肠道微生物的影响

发布时间：2018-04-19 09:56

本文选题：溶菌酶 + 断奶仔猪　；参考：《四川农业大学》2015年硕士论文

【摘要】：溶菌酶(Lysozyme,简写：LysM)是一种能水解细菌细胞壁的水解酶。作为具有抗菌作用的蛋白酶,溶菌酶是一种潜在的安全、高效饲用抗生素替代品。研究证明,饲用溶菌酶可提高断奶仔猪生长性能,改善仔猪肠道健康。但溶菌酶在断奶仔猪饲粮中的最适添加剂量和替代饲用抗生素的所需最低剂量还不确定,溶菌酶对大肠杆菌攻毒仔猪肠道的作用效果理论数据较少。本论文旨在研究不同水平溶菌酶对断奶仔猪生长性能、肠道形态和免疫功能的影响,以及溶菌酶对大肠杆菌攻毒仔猪肠道形态和肠道微生物的影响。试验一饲粮添加不同水平溶菌酶对断奶仔猪生长性能、肠道形态和免疫功能的影响试验采用单因子设计,共选取150头25日龄断奶的DLY仔猪(体重：7.32±0.90kg),随机分为6个处理组,每个处理组5个重复,每个重复5头仔猪。分别饲喂在玉米-豆粕型基础饲粮中添加20 mg/kg硫酸抗敌素+50 mg/kg吉他霉素,0、30、60、90、120 mg/kg溶菌酶的试验饲粮。试验期为28天,试验第14、28天采集仔猪血液样品,用于测定血清总补体和溶菌酶活性,以及血液中性粒细胞吞噬活性。试验结束时,分别从对照组,抗生素组,60、90mg/kg溶菌酶组各选出4头仔猪进行屠宰,收集仔猪腹腔巨噬细胞用于测定巨噬细胞吞噬活性,采集仔猪十二指肠、空肠、回肠组织,用于肠道形态测定及基因分析。结果表明：(1)试验0-14天,各处理间仔猪平均日增重无显著差异(P0.05)。试验14～28天和0-28天,90、120 mg/kg溶菌酶组及抗生素组仔猪平均日增重均显著高于对照组(P0.05)。在试验各阶段,各处理间仔猪平均日采食量和料重比均无显著差异(P0.05)。(2)各处理间回肠绒毛高度、隐窝深度及绒隐比均无显著差异(P0.05)。抗生素组,60、90 mg/kg溶菌酶组仔猪十二指肠绒毛高度及绒隐比均显著高于对照组(P0.05)。抗生素组和90 mg/kg溶菌酶组仔猪空肠隐窝深度显著低于对照组(P0.05),绒隐比显著高于对照组(P0.05)。(3)各处理间仔猪血清总补体和溶菌酶活性均无显著差异(P0.05)。抗生素组,60、90 mg/kg溶菌酶组仔猪腹腔巨噬细胞吞噬活性均显著高于对照组(P0.05)。(4)各处理间回肠TNF-α、IFN-γ、IL-8和TGF-β的mRNA表达量均无显著差异(P0.05)。试验二饲粮添加溶菌酶对大肠杆菌攻毒仔猪生长性能、肠道形态和肠道微生物的影响试验采用2×2因子试验设计,两个因素分别为添加溶菌酶和大肠杆菌攻毒。选取DLY断奶仔猪(体重：6.62±0.15)60头,随机分为4个处理组(C-E.Coli、C+E.Coli、 L-E.Coli、L+E.Coli),每个处理组15个重复,每个重复1头仔猪。C-E. Coli和C+E.Coli组饲喂基础饲粮,L-E.Coli和L+E.Coli组饲喂在基础饲粮中添加100 mg/kg溶菌酶的试验饲粮。仔猪断奶当天开始正式试验,记为第0天,试验期共14天。试验第11天早上对所有仔猪进行空腹称重后,对C+E.Coli和L+E.Coli组所有仔猪经口腔灌服20 mL大肠杆菌液(E.Coli, 1×108CFU/mL),24小时后重复攻毒一次,C-E.Coli和L-E.Coli组所有仔猪经口腔灌服相同剂量的无菌培养液作为对照。试验第14天早上进行屠宰采样,测定仔猪肠道形态和肠道微生物。结果表明：(1)大肠杆菌攻毒和饲粮溶菌酶对仔猪平均日增重、平日采食量、料重比和仔猪腹泻评分均无显著影响(P0.05)。(2)大肠杆菌攻毒增加仔猪空肠和回肠隐窝深度(P0.05)。饲粮溶菌酶降低仔猪空肠隐窝深度(P0.05),增加仔猪回肠绒毛高度和十二指肠、空肠、回肠绒隐比(P0.05)。L-E.Coli组仔猪空肠隐窝深度低于C-E.Coli组(P0.05),回肠绒毛高度和绒隐比高于C-E.Coli组(P0.05)。L+E.Coli组仔猪十二指肠和回肠隐窝深度低于C+E.Coli组,回肠绒隐比高于C+E.Coli组。(3)大肠杆菌攻毒增加仔猪回肠和结肠大肠杆菌数(P0.05),降低回肠和结肠乳酸杆菌和双歧杆菌数(P0.05)。饲粮溶菌酶降低仔猪回肠和结肠大肠杆菌数(P0.05),增加回肠和结肠乳酸杆菌和双歧杆菌数(P0.05)。L-E.Coli组仔猪回肠大肠杆菌数低于C-E.Coli组(P0.05),回肠和结肠乳酸杆菌及双歧杆菌数均高于C-E.Coli 鍣(p0.05)。L+E.Coli组仔猪回肠和结肠大肠杆菌数均低于C+E.Coli组(P0.05),乳酸杆菌均高于C+E.Coli组(P0.05),且回肠双歧杆菌数高于C+E.Coli组(P0.05)。根据以上结果,得出以下结论：(1)断奶仔猪饲粮中添加溶菌酶可改善肠道形态,增强免疫力,促进生长。(2)饲粮添加溶菌酶可改善大肠杆菌攻毒仔猪肠道形态和微生物结构,缓解仔猪免疫应激。(3)断奶仔猪饲粮添加90 mg/kg溶菌酶可达到与抗生素(20 mg/kg硫酸抗敌素+50 mg/kg吉他霉素)相同的促生长效果。
[Abstract]:Lysozyme (Lysozyme, abbreviation: LysM) is a kind of hydrolysis of the bacterial cell wall hydrolase. As antibacterial protease, lysozyme is a potentially safe, efficient feed antibiotic alternatives. Studies have shown that lysozyme can improve the growth performance of weaned piglets, weaned piglets intestinal health. But lysozyme in weaned piglets the optimum adding dose and alternative antibiotics required minimum dose is uncertain, the effect of the theory of data less lysozyme on Escherichia coli challenged piglets. This thesis aims to study the different levels of lysozyme on growth performance of weaned piglets. The effects of morphology and intestinal immune function, and the effect of lysozyme on Escherichia coli challenged piglets the morphology and intestinal microflora. The experiment of adding a different dietary levels of lysozyme on growth performance of weaned piglets, morphology and intestinal immune function The single factor design experiment with effect, a total of 150 25 day old weaned piglets DLY (weight: 7.32 + 0.90kg), were randomly divided into 6 groups, each group 5 repeats, each repeat. 5 piglets were fed with 20 mg/kg sulfuric acid in the corn soybean basal diet against the enemy in +50 mg/kg kitasamycin, test of dietary 0,30,60,90120 mg/kg lysozyme. The experiment period was 28 days, the test day 14,28 collected blood samples for piglets, determination of serum total complement and lysozyme activity and blood neutrophil phagocytic activity. At the end of the experiment, respectively from the control group, antibiotic group, each group selected 4 head 60,90mg/kg lysozyme the piglets were slaughtered, were collected for the determination of macrophage phagocytic activity of peritoneal macrophages, collection of duodenum, jejunum, ileum, and gene analysis for the determination of intestinal morphology. The results showed that: (1) test for 0-14 days, Among the treatments ADG had no significant difference (P0.05). The test for 14~28 days and 0-28 days, 90120 mg/kg lysozyme group and antibiotic group ADG were significantly higher than those in control group (P0.05). In each phase of the test, the piglets average daily feed intake and feed conversion ratio were not significantly different (P0.05). (2) the treatment between ileal villus height, crypt depth and velvet hidden than there was no significant difference (P0.05). The antibiotic group, 60,90 mg/kg group of duodenum villus height and lysozyme hair implicit were higher than that of control group (P0.05). The antibiotic group and 90 mg/kg lysozyme group were significantly lower than the control crypt depth of jejunum group (P0.05), hair implicit than was significantly higher than the control group (P0.05). (3) the total serum complement and lysozyme activity had no significant difference (P0.05). The antibiotic group, 60,90 mg/kg group of piglets lysozyme macrophage phagocytic activity were significantly Higher than that of the control group (P0.05). (4) among the treatments with ileal TNF- alpha, IFN- gamma, IL-8 and TGF- beta mRNA expression showed no significant difference (P0.05) test. Two dietary lysozyme on Escherichia coli challenged piglets growth performance, intestinal morphology and intestinal microbial effects of 2 * 2 factorial design uses the test, two factors were added to lysozyme and Escherichia coli challenged. Selected DLY of weaned piglets (weight: 6.62 + 0.15) 60, were randomly divided into 4 treatment groups (C-E.Coli, C+E.Coli, L-E.Coli, L+E.Coli), each treatment consisted of 15 replications, 1 piglets per replicate.C-E. Coli and C+E.Coli were fed the basal diet, experimental diets L-E.Coli and L+E.Coli were fed with 100 mg/kg lysozyme in the basal diet. Weaning day began formal test, write for zeroth days, the test period of 14 days. On the morning of the eleventh day test all piglets were fasting after weighing, on C+E.Coli and L+E. Coli all piglets by oral gavage group 20 Escherichia coli mL solution (E.Coli, 1 * 108CFU/mL), 24 hours after repeated inoculation time, all piglets of C-E.Coli and L-E.Coli group were orally treated by sterile culture medium as control. The morning of the fourteenth day of slaughter test sampling, determination of piglet intestinal morphology and intestinal tract microbial. The results showed that: (1) the Escherichia coli challenged and dietary lysozyme on ADG, daily intake, have no significant effect on feed weight ratio and diarrhea score (P0.05). (2) the Escherichia coli challenged piglets increased jejunal and ileal crypt depth (P0.05). Dietary lysozyme decrease piglet jejunum the depth (P0.05), increased piglet ileal villus height and duodenum, jejunum, ileum hair implicit (P0.05) than group.L-E.Coli piglet jejunal crypt depth was lower than that of C-E.Coli group (P0.05), ileum and cashmere hidden ratio is higher than C-E.Coli group (P0.05.L+E.C) Group oli piglet duodenum and ileum crypt depth is lower than the C+E.Coli group, the ileum ratio is higher than C+E.Coli group. The hidden pile (3) of Escherichia coli infection increased the ileum and colon of Escherichia coli in piglets (P0.05), reduce the number of ileum and colon of Lactobacillus and Bifidobacterium number (P0.05). Dietary lysozyme decreased ileum and colon Escherichia coli in piglets (P0.05), increase the ileum and colon of Lactobacillus and Bifidobacterium number (P0.05) of.L-E.Coli group were lower than that of group C-E.Coli in ileum of Escherichia coli (P0.05), ileum and colon of Lactobacillus and Bifidobacterium were higher than that of C-E.Coli Zhao (P0.05) group.L+E.Coli piglet ileum and colon Escherichia coli were lower than those in group C+E.Coli (P0.05). Lactic acid bacteria were higher than that of group C+E.Coli (P0.05), and the number of Bifidobacterium ileum was higher than C+E.Coli group (P0.05). According to the above results, draw the following conclusions: (1) adding lysozyme weaned pig diet can improve bowel shape State, enhance immunity, promote growth. (2) dietary lysozyme can improve the Escherichia coli of intestinal morphology and microbial structure of poison piglets, alleviate the piglet immunological stress. (3) diet of weaned piglets supplemented with 90 mg/kg lysozyme can be reached with antibiotics (20 mg/kg colistin sulfate +50 mg/kg kitasamycin) the growth promoting effect of the same.

【学位授予单位】：四川农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S828.5

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