發(fā)布時(shí)間：2018-04-19 09:56

  本文選題：溶菌酶 + 斷奶仔豬　； 參考：《四川農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：溶菌酶(Lysozyme,簡(jiǎn)寫：LysM)是一種能水解細(xì)菌細(xì)胞壁的水解酶。作為具有抗菌作用的蛋白酶,溶菌酶是一種潛在的安全、高效飼用抗生素替代品。研究證明,飼用溶菌酶可提高斷奶仔豬生長(zhǎng)性能,改善仔豬腸道健康。但溶菌酶在斷奶仔豬飼糧中的最適添加劑量和替代飼用抗生素的所需最低劑量還不確定,溶菌酶對(duì)大腸桿菌攻毒仔豬腸道的作用效果理論數(shù)據(jù)較少。本論文旨在研究不同水平溶菌酶對(duì)斷奶仔豬生長(zhǎng)性能、腸道形態(tài)和免疫功能的影響,以及溶菌酶對(duì)大腸桿菌攻毒仔豬腸道形態(tài)和腸道微生物的影響。試驗(yàn)一飼糧添加不同水平溶菌酶對(duì)斷奶仔豬生長(zhǎng)性能、腸道形態(tài)和免疫功能的影響試驗(yàn)采用單因子設(shè)計(jì),共選取150頭25日齡斷奶的DLY仔豬(體重：7.32±0.90kg),隨機(jī)分為6個(gè)處理組,每個(gè)處理組5個(gè)重復(fù),每個(gè)重復(fù)5頭仔豬。分別飼喂在玉米-豆粕型基礎(chǔ)飼糧中添加20 mg/kg硫酸抗敵素+50 mg/kg吉他霉素,0、30、60、90、120 mg/kg溶菌酶的試驗(yàn)飼糧。試驗(yàn)期為28天,試驗(yàn)第14、28天采集仔豬血液樣品,用于測(cè)定血清總補(bǔ)體和溶菌酶活性,以及血液中性粒細(xì)胞吞噬活性。試驗(yàn)結(jié)束時(shí),分別從對(duì)照組,抗生素組,60、90mg/kg溶菌酶組各選出4頭仔豬進(jìn)行屠宰,收集仔豬腹腔巨噬細(xì)胞用于測(cè)定巨噬細(xì)胞吞噬活性,采集仔豬十二指腸、空腸、回腸組織,用于腸道形態(tài)測(cè)定及基因分析。結(jié)果表明：(1)試驗(yàn)0-14天,各處理間仔豬平均日增重?zé)o顯著差異(P0.05)。試驗(yàn)14～28天和0-28天,90、120 mg/kg溶菌酶組及抗生素組仔豬平均日增重均顯著高于對(duì)照組(P0.05)。在試驗(yàn)各階段,各處理間仔豬平均日采食量和料重比均無(wú)顯著差異(P0.05)。(2)各處理間回腸絨毛高度、隱窩深度及絨隱比均無(wú)顯著差異(P0.05)。抗生素組,60、90 mg/kg溶菌酶組仔豬十二指腸絨毛高度及絨隱比均顯著高于對(duì)照組(P0.05)。抗生素組和90 mg/kg溶菌酶組仔豬空腸隱窩深度顯著低于對(duì)照組(P0.05),絨隱比顯著高于對(duì)照組(P0.05)。(3)各處理間仔豬血清總補(bǔ)體和溶菌酶活性均無(wú)顯著差異(P0.05)�？股亟M,60、90 mg/kg溶菌酶組仔豬腹腔巨噬細(xì)胞吞噬活性均顯著高于對(duì)照組(P0.05)。(4)各處理間回腸TNF-α、IFN-γ、IL-8和TGF-β的mRNA表達(dá)量均無(wú)顯著差異(P0.05)。試驗(yàn)二飼糧添加溶菌酶對(duì)大腸桿菌攻毒仔豬生長(zhǎng)性能、腸道形態(tài)和腸道微生物的影響試驗(yàn)采用2×2因子試驗(yàn)設(shè)計(jì),兩個(gè)因素分別為添加溶菌酶和大腸桿菌攻毒。選取DLY斷奶仔豬(體重：6.62±0.15)60頭,隨機(jī)分為4個(gè)處理組(C-E.Coli、C+E.Coli、 L-E.Coli、L+E.Coli),每個(gè)處理組15個(gè)重復(fù),每個(gè)重復(fù)1頭仔豬。C-E. Coli和C+E.Coli組飼喂基礎(chǔ)飼糧,L-E.Coli和L+E.Coli組飼喂在基礎(chǔ)飼糧中添加100 mg/kg溶菌酶的試驗(yàn)飼糧。仔豬斷奶當(dāng)天開(kāi)始正式試驗(yàn),記為第0天,試驗(yàn)期共14天。試驗(yàn)第11天早上對(duì)所有仔豬進(jìn)行空腹稱重后,對(duì)C+E.Coli和L+E.Coli組所有仔豬經(jīng)口腔灌服20 mL大腸桿菌液(E.Coli, 1×108CFU/mL),24小時(shí)后重復(fù)攻毒一次,C-E.Coli和L-E.Coli組所有仔豬經(jīng)口腔灌服相同劑量的無(wú)菌培養(yǎng)液作為對(duì)照。試驗(yàn)第14天早上進(jìn)行屠宰采樣,測(cè)定仔豬腸道形態(tài)和腸道微生物。結(jié)果表明：(1)大腸桿菌攻毒和飼糧溶菌酶對(duì)仔豬平均日增重、平日采食量、料重比和仔豬腹瀉評(píng)分均無(wú)顯著影響(P0.05)。(2)大腸桿菌攻毒增加仔豬空腸和回腸隱窩深度(P0.05)。飼糧溶菌酶降低仔豬空腸隱窩深度(P0.05),增加仔豬回腸絨毛高度和十二指腸、空腸、回腸絨隱比(P0.05)。L-E.Coli組仔豬空腸隱窩深度低于C-E.Coli組(P0.05),回腸絨毛高度和絨隱比高于C-E.Coli組(P0.05)。L+E.Coli組仔豬十二指腸和回腸隱窩深度低于C+E.Coli組,回腸絨隱比高于C+E.Coli組。(3)大腸桿菌攻毒增加仔豬回腸和結(jié)腸大腸桿菌數(shù)(P0.05),降低回腸和結(jié)腸乳酸桿菌和雙歧桿菌數(shù)(P0.05)。飼糧溶菌酶降低仔豬回腸和結(jié)腸大腸桿菌數(shù)(P0.05),增加回腸和結(jié)腸乳酸桿菌和雙歧桿菌數(shù)(P0.05)。L-E.Coli組仔豬回腸大腸桿菌數(shù)低于C-E.Coli組(P0.05),回腸和結(jié)腸乳酸桿菌及雙歧桿菌數(shù)均高于C-E.Coli 鍣(p0.05)。L+E.Coli組仔豬回腸和結(jié)腸大腸桿菌數(shù)均低于C+E.Coli組(P0.05),乳酸桿菌均高于C+E.Coli組(P0.05),且回腸雙歧桿菌數(shù)高于C+E.Coli組(P0.05)。根據(jù)以上結(jié)果,得出以下結(jié)論：(1)斷奶仔豬飼糧中添加溶菌酶可改善腸道形態(tài),增強(qiáng)免疫力,促進(jìn)生長(zhǎng)。(2)飼糧添加溶菌酶可改善大腸桿菌攻毒仔豬腸道形態(tài)和微生物結(jié)構(gòu),緩解仔豬免疫應(yīng)激。(3)斷奶仔豬飼糧添加90 mg/kg溶菌酶可達(dá)到與抗生素(20 mg/kg硫酸抗敵素+50 mg/kg吉他霉素)相同的促生長(zhǎng)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.

【學(xué)位授予單位】：四川農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S828.5

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