新型氧化锌对断奶仔猪生产性能与肠道功能的影响及其作用机制研究

发布时间：2018-04-17 05:12

本文选题：新型氧化锌 + 表征　；参考：《浙江大学》2015年博士论文

【摘要】：断奶是仔猪最为重要的阶段。环境、营养和心理等应激伴随微生物、免疫和生理等变化共同作用于未成熟的消化道,可导致断奶仔猪腹泻、生长性能降低。在2006年欧盟禁用后,许多营养手段(如氧化锌、益生菌、益生元、有机酸和精油等)广泛用于替代抗生素用于断奶仔猪防腹泻、促生长。药理剂量(2000-4000mg/kg)氧化锌用于断奶仔猪已有二十多年历史。然而,高剂量氧化锌带来动物毒性、细菌抗性和环境污染等问题迫使欧盟颁布150 mg/kg的最高限量。因此,亟需一种替代产品,减少氧化锌用量、降低粪锌排放。由于作用机制不同,上述营养手段和不同锌源并不能完全替代氧化锌。其他形式氧化锌(如纳米氧化锌和氧化锌-天然硅酸盐复合物等)虽有一定效果,但仍存在效果不稳定与安全风险等问题。由于氧化锌活性与其比表面积正相关,而毒性与其粒径负相关,本研究提出“高比表面积-微米粒径”的新型氧化锌研制思路,制备并表征了新型氧化锌,研究了其溶解性与生物活性,还研究了其对断奶仔猪生产性能与肠道功能的影响及其作用机制,结果如下：1新型氧化锌的制备、理化性质与生物活性根据前驱体碱式碳酸锌热重分析结果,辅以适宜加热速率和时间,制备得到了新型氧化锌,并使用扫描电子显微镜、透射电子显微镜、BET和X-射线衍射等方法对其进行表征。结果表明,新型氧化锌比表面积约48.9 m2/g、粒径约10～30μm,内部为由小梁支撑的中空结构,并且小梁含有丰富的孔腔。与普通氧化锌、纳米氧化锌和多孔氧化锌相比,新型氧化锌对于人工胃液分解具有更好的保护作用；对大肠杆菌K88、肠炎沙门氏菌和金黄色葡萄球菌具有更好的抗菌活性,使大肠杆菌K88变短且表面呈浆膜样；对猪小肠上皮细胞IPEC-1具有更强的增殖促进作用和更低的细胞毒性,但对其内部活性氧水平无影响。提示,新型氧化锌的这些特性可能与其特殊结构有关。2新型氧化锌对断奶仔猪生长性能与腹泻的影响本研究共进行三次饲养试验,分别对比研究不同形式氧化锌、不同剂量新型氧化锌以及新型氧化锌与不同添加剂配伍,对断奶仔猪日增重、日采食量、饲料转化率和腹泻率等的影响。试验Ⅰ,56头“杜×长×黑”断奶仔猪(25±1 d,7.33 kg)分为4组：基础日粮+3000 mg/kg普通氧化锌、基础日粮+300 mg/kg纳米氧化锌、基础日粮+300 mg/kg多孔氧化锌和基础日粮+300 mg/kg新型氧化锌。试验Ⅱ,165头“杜×长”断奶仔猪(21±1d,6.40 kg)分为5组：基础日粮+3000 mg/kg普通氧化锌、基础日粮+300 mg/kg普通氧化锌、基础日粮+600 mg/kg新型氧化锌、基础日粮+300 mg/kg新型氧化锌和基础日粮+150 mg/kg新型氧化锌。试验Ⅲ,90头“杜x长×大”断奶仔猪(25±1 d,7.88 kg)分为3组：基础日粮+3000 mg/kg普通氧化锌+抗生素(“高剂量普通氧化锌+抗生素”组)、基础日粮+300 mg/kg新型氧化锌+抗生素(“低剂量新型氧化锌+抗生素”组)和基础日粮+300 mg/kg新型氧化锌+益生菌(“低剂量新型氧化锌+益生菌”组)。结果表明,300 mg/kg新型氧化锌防腹泻、促生长效果不同程度优于3000 mg/kg普通氧化锌和300 mg/kg其他形式氧化锌(纳米氧化锌或多孔氧化锌)；与3000 mg/kg普通氧化锌相比,不同剂量新型氧化锌(600或300 mg/kg)以及300 mg/kg新型氧化锌与不同添加剂(抗生素或益生菌)配伍均可有效降低断奶仔猪腹泻、促进断奶仔猪生长。3新型氧化锌对断奶仔猪养分消化与锌吸收转运的影响与高剂量普通氧化锌相比,低剂量新型氧化锌可以提高养分表观消化率和十二指肠内容物消化酶活性；极显著(P0.01)降低肝脏中锌的沉积和粪便中锌的排放,但对血清中锌的水平影响不显著(P0.05)；显著(P0.05)提高空肠和回肠锌转运蛋白ZIP4基因表达水平,但对锌转运蛋白ZnT1、ZnT2和ZnT5的影响较小；并极显著(P0.01)降低十二指肠、空肠、回肠、结肠和肝脏金属硫蛋白MT1基因表达水平,提高血清中超氧化物歧化酶和过氧化氢酶活性,极显著(P0.01)提高血清中总抗氧化力水平。4新型氧化锌对断奶仔猪肠道形态与屏障功能的影响与高剂量普通氧化锌相比,低剂量新型氧化锌可以改善消化道形态,极显著(P0.01)提高十二指肠绒毛高度：隐窝深度和空肠微绒毛高度；提高空肠和回肠紧密连接蛋白claudin-1、occludin、ZO-1和ZO-2基因表达水平；提高回肠组织、回肠黏膜和回肠黏液的sIgA水平；提高空肠和回肠粘蛋白MUC1、MUC2、MUC4、MUC13和MUC20基因表达水平；提高回肠微生物区系的丰度和多样性。配伍益生菌还可以提高空肠、回肠和结肠抗菌肽pBD-1和pBD-2基因表达水平。综上所述,本研究成功研制了“高比表面积-微米粒径”新型氧化锌,使其具有高抗菌活性与低细胞毒性。饲养试验发现,与高剂量(3000 mg/kg)普通氧化锌相比,低剂量(300 mg/kg)新型氧化锌可以有效降低断奶仔猪腹泻、促进断奶仔猪生长、提高养分表观消化率、抵抗氧化应激,并可有效改善肠道形态、增强肠道屏障功能。
[Abstract]:Is the most important stage of weaning piglets. Environment, nutrition and psychological stress with microorganisms, interaction of immune and physiological changes of the digestive tract in immature, can cause diarrhea of weaned piglets, growth performance decreased. In 2006 the EU banned, many nutrition means (such as Zinc Oxide, probiotics, prebiotics, organic acids and essential oils etc.) is widely used for replacing antibiotics for the prevention of diarrhea in weaned piglets, growth at pharmacological doses (2000-4000mg/kg) for Zinc Oxide has been weaned piglets more than 20 years of history. However, the high dose of Zinc Oxide brings animal toxicity, bacteria resistance and environmental pollution has forced the highest limit issued 150 mg/kg. Therefore, we need a replacement product, reduce Zinc Oxide the dosage, reduce fecal zinc emissions. Due to different mechanisms, the nutrition means and different zinc sources can not completely replace Zinc Oxide. Other forms (such as Zinc Oxide Zinc Oxide and Zinc Oxide - natural silicate nanometer compound etc.) has a certain effect, but there are still unstable effect and security risks and other issues. Due to Zinc Oxide's activity and the specific surface area is related to toxicity and particle size are negatively correlated, this research proposes the specific surface area development of new Zinc Oxide - micron size "ideas, preparation and characterization of a new Zinc Oxide, studied the solubility and biological activity, but also to study their impact on piglet performance and intestinal function and its mechanism, the results are as follows: 1 new Zinc Oxide preparation, the precursor basic zinc carbonate thermogravimetric analysis results and chemical properties and biological activity, supplemented by appropriate heating the rate and time of preparation of the new Zinc Oxide, and using scanning electron microscopy, transmission electron microscopy, BET and X- ray diffraction method were characterized. The results show that the new type of oxygen Zinc surface area of about 48.9 m2/g, diameter 10~30 m, inside the hollow structure by small beam, and beam containing cavity. The rich and ordinary compared to Zinc Oxide and Zinc Oxide, nano porous Zinc Oxide, Zinc Oxide model for decomposition of artificial gastric juice has better protective effect; on Escherichia coli K88, Salmonella enteritidis Salmonella and Staphylococcus aureus has better antibacterial activity to Escherichia coli K88 becomes shorter and the surface is more serous; porcine small intestine epithelial cells IPEC-1 proliferation promoting cell toxicity and lower, but had no effect on the internal level of reactive oxygen species. The results indicated that the effects of these characteristics may the new Zinc Oxide.2 with the special structure of the new Zinc Oxide on growth performance and diarrhea of weaned piglets in this study were three times feeding test, respectively comparative study of different forms of Zinc Oxide, different doses of new oxidation The new Zinc Oxide and zinc with different additives, the piglets daily gain, daily feed intake, feed conversion rate and the rate of diarrhea and so on. In Experiment I, 56 "Duroc * Landrace * BLACK weaned piglets (25 + 1 D, 7.33 kg) were divided into 4 groups: basal diet +3000 mg/kg Zinc Oxide, basal diet +300 mg/kg nano diet +300 Zinc Oxide, Zinc Oxide and mg/kg porous diet +300 mg/kg model. The Zinc Oxide II test, 165 crossbred weaned piglets * long (21 + 1D, 6.40 kg) were divided into 5 groups: basal diet +3000 mg/kg common Zinc Oxide, basal diet + 300 mg/kg general Zinc Oxide, basal diet +600 mg/kg new Zinc Oxide, basal diet +300 mg/kg model and Zinc Oxide based diet +150 mg/kg new Zinc Oxide. Test III, 90" long Du x * "weaned piglets (25 + 1 D, 7.88 kg) were divided into 3 groups: basal diet +3000 mg/kg general + Zinc Oxide antibiotics (" High dose ordinary Zinc Oxide + antibiotic group), basal diet +300 mg/kg new antibiotics ("Zinc Oxide + Zinc Oxide + low dose of new antibiotics" group) and the basal diet +300 mg/kg new Zinc Oxide + probiotics ("low dose model Zinc Oxide + probiotics group). The results showed that the 300 mg/kg new Zinc Oxide anti diarrhea, growth the effect of different levels of less than 3000 mg/kg mg/kg and 300 other forms of ordinary Zinc Oxide Zinc Oxide (Zinc Oxide or Zinc Oxide; nano porous) compared with 3000 mg/kg Zinc Oxide, Zinc Oxide model of different doses (600 or 300 mg/kg) and 300 mg/kg new Zinc Oxide and different additives (antibiotics or probiotics) and can effectively reduce the diarrhea of weaned piglets, promote the growth of weaned piglets the new Zinc Oxide.3 on nutrient digestion and absorption of zinc in weaned piglets compared with high dose of ordinary Zinc Oxide, low dose model oxidation Zinc can improve the nutrient apparent enzyme activity rate and duodenal contents digest; (P0.01) significantly decrease the deposition of zinc in liver and fecal zinc emissions, but the effect on serum zinc levels was not significant (P0.05); (P0.05) significantly improve the jejunum and ileum of zinc transporter ZIP4 gene expression level. But the zinc transporter ZnT1, little effect on ZnT2 and ZnT5; and (P0.01) significantly reduced the duodenum, jejunum, ileum, colon and liver metallothionein gene expression level of MT1, increase serum superoxide dismutase and catalase activity significantly (P0.01) to improve the effect of serum total antioxidant capacity level.4 the new Zinc Oxide of piglet intestinal morphology and barrier function compared with the ordinary high dose low dose model of Zinc Oxide, Zinc Oxide can improve the digestive tract morphology, significantly increased by twelve (P0.01) refers to the villus height: hidden Pit depth and villus height of jejunum; improve the tight junction protein claudin-1, occludin in jejunum and ileum, ZO-1 and ZO-2 gene expression level; improve the ileal tissue, ileum mucosa and ileum mucous sIgA level; improve the jejunal and ileal mucin MUC1, MUC2, MUC4, MUC13 and MUC20 gene expression level; improve the abundance of ileal microflora the diversity and compatibility. Probiotics can also improve the jejunum, ileum and colon of antibacterial peptide pBD-1 and pBD-2 gene expression. In summary, this study successfully developed "high surface area - micron particle size" model of Zinc Oxide, which has high antibacterial activity and low cytotoxicity. The feeding experiment and found that high dose (3000 mg/kg) compared with ordinary Zinc Oxide, low dose (300 mg/kg) Zinc Oxide model can effectively reduce the diarrhea of weaned piglets, weaned piglets to promote growth, improve the apparent digestibility of nutrients, resistance to oxidative stress It can effectively improve the intestinal morphology and enhance the intestinal barrier function.

【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：S828.5

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