WO2024221685A1 - 一种饲料级氧化锌的制备及其在断奶仔猪上的应用 - Google Patents

一种饲料级氧化锌的制备及其在断奶仔猪上的应用 Download PDF

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WO2024221685A1
WO2024221685A1 PCT/CN2023/116022 CN2023116022W WO2024221685A1 WO 2024221685 A1 WO2024221685 A1 WO 2024221685A1 CN 2023116022 W CN2023116022 W CN 2023116022W WO 2024221685 A1 WO2024221685 A1 WO 2024221685A1
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zinc oxide
feed
weaned piglets
grade
grade zinc
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汪以真
苏维发
路则庆
李治学
靳明亮
王凤芹
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Zhejiang University
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Zhejiang University
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    • C—CHEMISTRY; METALLURGY
    • C01—INORGANIC CHEMISTRY
    • C01G—COMPOUNDS CONTAINING METALS NOT COVERED BY SUBCLASSES C01D OR C01F
    • C01G9/00—Compounds of zinc
    • C01G9/02—Oxides; Hydroxides
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/02—Breeding vertebrates
    • C—CHEMISTRY; METALLURGY
    • C01—INORGANIC CHEMISTRY
    • C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
    • C01P2004/00—Particle morphology
    • C01P2004/01—Particle morphology depicted by an image
    • C01P2004/04—Particle morphology depicted by an image obtained by TEM, STEM, STM or AFM
    • C—CHEMISTRY; METALLURGY
    • C01—INORGANIC CHEMISTRY
    • C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
    • C01P2004/00—Particle morphology
    • C01P2004/60—Particles characterised by their size
    • C01P2004/61—Micrometer sized, i.e. from 1-100 micrometer
    • C—CHEMISTRY; METALLURGY
    • C01—INORGANIC CHEMISTRY
    • C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
    • C01P2006/00—Physical properties of inorganic compounds
    • C01P2006/12—Surface area

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  • the invention belongs to the technical field of animal feed, and in particular relates to a preparation method of feed-grade zinc oxide and application thereof on weaned piglets.
  • the piglet stage is one of the most important links in the pig farming process.
  • Early weaning stress in piglets can cause intestinal microbial disorders, mucosal immune damage and nutrient absorption disorders, which in turn lead to decreased production performance, diarrhea, and even death.
  • Data show that the mortality rate in the piglet stage accounts for more than 85% of the entire pig farming process, causing significant economic losses to the pig farming industry.
  • the addition of feed antibiotics and related drugs was one of the main measures to prevent and treat piglet weaning stress syndrome, but it would lead to the production of drug-resistant strains and drug residues in animal-derived foods, which had many adverse effects on human health and the environment.
  • zinc oxide, probiotics, prebiotics, antimicrobial peptides, organic acids and essential oils were used as antibiotic substitutes to alleviate diarrhea in weaned piglets and improve piglet growth performance.
  • the purpose of the present invention is to provide a method for preparing a new type of feed-grade zinc oxide and its application in weaned piglets.
  • the specific surface area of zinc oxide is increased, the antibacterial activity is enhanced, its dissolution in the acidic environment of the stomach and its toxicity to pig intestinal epithelial cells are reduced, and the bioavailability of zinc in piglets is improved, thereby solving the problems of animal toxicity, bacterial resistance and environmental pollution caused by the application of high-dose zinc oxide in weaned piglets.
  • a method for preparing feed-grade zinc oxide comprises the following steps: using basic zinc carbonate as a preparation raw material, calcining the basic zinc carbonate in a muffle furnace at a preparation temperature of 250-300 DEG C for 2 hours, and preparing feed-grade zinc oxide.
  • a feed-grade zinc oxide obtained according to the preparation method with a specific surface area of 48.9m2 /g and a particle size of 10 to 30 ⁇ m,
  • the microstructure obtained by transmission electron microscopy analysis is a hollow structure supported by trabeculae, and the trabeculae contain abundant cavities.
  • the feed-grade zinc oxide is not easily decomposed in artificial gastric juice, has better antibacterial activity against Escherichia coli K88, Salmonella enteritidis CMCC5004 and Staphylococcus aureus ATCC25923, and has lower cytotoxicity to porcine small intestinal epithelial cells IPEC-1.
  • the application of the feed-grade zinc oxide on weaned piglets is to add a low dose of 300 mg/kg feed-grade zinc oxide to the basic daily diet of the weaned piglets to reduce the diarrhea rate of the weaned piglets.
  • the application of the combination of feed-grade zinc oxide and probiotics on weaned piglets is that low-dose 300 mg/kg feed-grade zinc oxide and probiotics 10 8 CFU/kg Clostridium butyricum ZJU-F1 and 10 9 CFU/kg Bacillus licheniformis are added to the basic diet of weaned piglets to reduce the diarrhea rate of weaned piglets, promote the growth of weaned piglets, improve the apparent digestibility of nutrients, reduce zinc discharge and liver deposition, improve the intestinal barrier and immune function of weaned piglets, and increase the abundance of intestinal probiotic Lactobacillus reuteri; the Clostridium butyricum ZJU-F1 (Clostridium butyricum) is an existing strain, and the preservation number of the General Microbiology Center of the China Microbiological Culture Collection Administration is CGMCC No: 8939; Bacillus licheniformis (Bacillus licheniformis) is purchased from Zhejiang Hu
  • the method has a simple preparation process, and the prepared new zinc oxide has a specific surface area of 48.9m2 /g and a particle size of 10-30 ⁇ m.
  • the prepared new zinc oxide is not easily decomposed in artificial gastric juice, has better antibacterial activity against Escherichia coli K88, Salmonella enteritidis and Staphylococcus aureus, and has lower cytotoxicity to porcine small intestinal epithelial cells IPEC-1.
  • high-dose ordinary zinc oxide low-dose nano zinc oxide and low-dose porous zinc oxide
  • low-dose new zinc oxide has a better effect of reducing the diarrhea rate of weaned piglets.
  • the combination of low-dose new zinc oxide and probiotics can effectively reduce the diarrhea rate of weaned piglets, promote the growth of weaned piglets, improve the apparent digestibility of nutrients, reduce zinc discharge, improve intestinal barrier and immune function, and increase the abundance of beneficial intestinal bacteria.
  • FIG. 1 is a transmission electron microscope image of basic zinc carbonate and zinc oxide prepared by the present invention.
  • FIG. 2 is a graph showing the solubility of zinc oxide prepared in the present invention in artificial gastrointestinal fluid.
  • FIG3 shows the inhibitory effect of zinc oxide prepared by the present invention on Escherichia coli K88, Salmonella enteritidis and Staphylococcus aureus.
  • FIG. 4 shows the toxic effect of zinc oxide prepared by the present invention on porcine intestinal epithelial cells IPEC-1.
  • FIG5 is the effect of zinc oxide prepared by the present invention and probiotics on the zinc content in the liver and feces of weaned piglets.
  • FIG6 is the effect of zinc oxide prepared by the present invention and probiotics on the jejunum morphology of weaned piglets.
  • FIG. 7 shows the effects of zinc oxide prepared by the present invention and probiotics on the intestinal barrier and immune function of weaned piglets.
  • FIG8 is the effect of zinc oxide prepared by the present invention and probiotics on the intestinal flora of weaned piglets.
  • a preparation method of a new feed-grade zinc oxide is as follows:
  • Basic zinc carbonate is used as a preparation raw material, and a thermogravimetric analysis curve of basic zinc carbonate is measured by a thermogravimetric analyzer.
  • the heating rate is 20°C/min
  • the heating temperature range is 50-500°C
  • the appropriate preparation temperature is 250-300°C.
  • basic zinc carbonate is calcined in a muffle furnace for 2 hours to prepare a feed-grade new zinc oxide with a particle size of 10-30 ⁇ m.
  • the prepared novel zinc oxide was analyzed by transmission electron microscopy, and its microstructure is shown in FIG1 .
  • the interior of the novel zinc oxide is a hollow structure supported by trabeculae, and the trabeculae contain abundant pores.
  • the activity of zinc oxide is positively correlated with its specific surface area.
  • the results are shown in Table 1.
  • the new zinc oxide has a larger specific surface area ( 48.92m2 /g) and total pore volume ( 0.57cm3 /g).
  • Artificial gastric juice (pH 3.8): 0.1mol/L HCl solution was adjusted to pH 3.8 with 1mol/L HCl solution and 1mol/L NaOH solution; artificial small intestinal juice (pH 6.8): 750ml 0.1mol/L HCl solution was mixed with 250ml 0.2mol/L Na3PO4 solution, and the pH was adjusted to 6.8 with 1mol/L HCl solution and 1mol/L NaOH solution; artificial colonic juice (pH 7.8): 750ml 0.1mol/L HCl solution was mixed with 250ml 0.2mol/L Na3PO4 solution, and the pH was adjusted to 7.8 with 1mol/L HCl solution and 1mol/L NaOH solution.
  • E. coli K88 is the main pathogen causing diarrhea in weaned piglets.
  • the new zinc oxide can not only shorten E. coli K88, but also destroy its surface structure.
  • the LDH release rate reflects the integrity of the cell membrane by detecting lactate dehydrogenase released into the cell during cell apoptosis or necrosis, and realizes quantitative analysis of cytotoxicity.
  • the new zinc oxide compared with ordinary zinc oxide, nano zinc oxide and porous zinc oxide, the new zinc oxide has the lowest LDH release rate, indicating that the new zinc oxide has the lowest cytotoxicity to porcine small intestinal epithelial cells.
  • the experiment selected 56 25-day-old weaned piglets (average weight 7.33kg, half male and half female) and divided them into 4 groups, 14 in each group, for 14 days.
  • the specific treatments are as follows: basal diet + 3000mg/kg ordinary zinc oxide group; basal diet + 300mg/kg nano zinc oxide group; basal diet + 300mg/kg porous zinc oxide group; basal diet + 300mg/kg new zinc oxide group.
  • basal diet + 3000mg/kg ordinary zinc oxide group basal diet + 300mg/kg nano zinc oxide group
  • basal diet + 300mg/kg new zinc oxide group basal diet + 300mg/kg new zinc oxide group.
  • the effect of new zinc oxide on the growth performance and diarrhea rate of weaned piglets was determined.
  • the effect of 300 mg/kg low-dose feed-grade new zinc oxide on promoting growth in weaned piglets is comparable to that of 3000 mg/kg high-dose ordinary zinc oxide.
  • 300 mg/kg nano zinc oxide and 300 mg/kg porous zinc oxide the addition of 300 mg/kg new zinc oxide to the basic diet of weaned piglets has the lowest diarrhea rate, indicating that low-dose new zinc oxide can be used as an effective alternative to high-dose ordinary zinc oxide for preventing and treating diarrhea in piglets.
  • the experiment selected 90 25-day-old weaned piglets (average 7.88 kg, half male and half female) and divided them into 3 groups, with 3 pens in each group and 10 pigs in each pen. The experiment lasted for 21 days.
  • the specific treatments were as follows: basal diet + 3000mg/kg ordinary zinc oxide + antibiotic group ("HOZO+Ant”group); basal diet + 300mg/kg new zinc oxide + antibiotic group ("LAZO+Ant”group); basal diet + 300mg/kg new zinc oxide + probiotic group (“LAZO+Pro” group).
  • Antibiotics 100mg/kg quinolone, 20mg/kg colistin sulfate and 50mg/kg kitasamycin; probiotics: 10 8 CFU/kg Clostridium butyricum ZJU-F1 and 10 9 CFU/kg Bacillus licheniformis.
  • the combination of 300mg/kg low-dose new zinc oxide and probiotics can basically achieve the anti-diarrhea and growth-promoting effects of 3000mg/kg high-dose ordinary zinc oxide and antibiotics, avoiding the pollution and other problems caused by high-dose zinc oxide.
  • low-dose novel zinc oxide combined with probiotics can effectively reduce zinc deposition in the liver of weaned piglets, effectively prevent high-dose zinc oxide from causing zinc overload in the liver of weaned piglets, and effectively reduce the pollution to the environment caused by zinc discharge in feces.
  • low-dose novel zinc oxide combined with probiotics can avoid the damage to the intestines of weaned piglets caused by high-dose ordinary zinc oxide and antibiotics, while increasing the length of intestinal microvilli and enhancing the intestinal absorption area.
  • low-dose novel zinc oxide combined with probiotics can increase the expression of intestinal tight junction proteins (Claudin-1 and Occludin), mucins (MUC1, MUC2, MUC13, MUC20) and antimicrobial peptides (pBD-1 and pBD-2) in weaned piglets, thereby improving the intestinal barrier and immune function of weaned piglets.
  • intestinal tight junction proteins Claudin-1 and Occludin
  • mucins MUC1, MUC2, MUC13, MUC20
  • pBD-1 and pBD-2 antimicrobial peptides
  • High-throughput sequencing was used to analyze the intestinal microbial flora of piglets. The results are shown in Figure 8.
  • Low-dose novel zinc oxide combined with probiotics increased the abundance of intestinal flora and the abundance of probiotic Lactobacillus reuteri in weaned piglets. It can promote animal intestinal digestion and absorption and inhibit the proliferation of pathogenic bacteria in the intestine.

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Abstract

本发明公开了一种饲料级氧化锌的制备及其在断奶仔猪上的应用。采用碱式碳酸锌作为制备原料，根据碱式碳酸锌的热重分析得到最佳煅烧温度250～300℃，将碱式碳酸锌在马弗炉中煅烧2h，制备得到饲料级新型氧化锌。该方法制备工艺简单，制备得到新型氧化锌比表面积48.9m2/g、粒径10～30μm。制备得到的新型氧化锌在人工胃液中不易被分解，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，对猪小肠上皮细胞IPEC-1具有更低的细胞毒性。将低剂量新型氧化锌和益生菌配伍可有效降低断奶仔猪腹泻率、促进断奶仔猪生长、提高养分表观消化率、减少锌排放，改善肠道屏障和免疫功能，提高肠道有益菌的丰度。

Description

一种饲料级氧化锌的制备及其在断奶仔猪上的应用 技术领域
本发明属于动物饲料技术领域，具体涉及一种饲料级氧化锌的制备方法及其在断奶仔猪上的应用。
背景技术
仔猪阶段是生猪养殖过程中最重要的环节之一，仔猪早期断奶应激会造成肠道微生物紊乱、黏膜免疫损伤和营养物质吸收障碍，进而导致生产性能下降、腹泻，甚至死亡。数据表明，仔猪阶段死亡率约占整个生猪养殖过程的85％以上，给生猪养殖产业带来重大的经济损失。饲料禁抗前，饲用抗生素和相关药物的添加是作为防治仔猪断奶应激综合征的主要措施之一，但会导致耐药菌株的产生及药物在动物源性食品中的残留等诸多问题，给人类健康及环境带来了诸多不利的影响。在饲料禁抗后，氧化锌、益生菌、益生元、抗菌肽、有机酸和精油等作为抗生素替代品被用于缓解断奶仔猪防腹泻和改善仔猪生长性能。
近年来，药理剂量的氧化锌作为断奶仔猪日粮添加剂得到了广泛应用，其具有促进仔猪生长、提高免疫力、降低腹泻等优点。但高剂量氧化锌在断奶仔猪上的应用也带来动物毒性、细菌抗性和环境污染等问题。2022年欧盟禁止了药理剂量的氧化锌在断奶后仔猪阶段使用。近年来，关于纳米氧化锌和多孔氧化锌制备的方法研究较多，比如CN106348336 A“一种纳米氧化锌及其制备方法”和CN112939059 A“一种多孔氧化锌的制备方法”，但其制备工艺相对复杂，且在仔猪上应用的系统研究较少，仍存在效果不稳定与安全风险等问题。
发明内容
为了克服现有技术的不足，本发明的目的是提供一种饲料级新型氧化锌的制备方法及其在断奶仔猪上的应用。通过制备新型氧化锌来提高氧化锌的比表面积、增强抗菌活性、降低其在胃中酸性环境下溶解以及对猪肠上皮细胞毒性，提高仔猪锌元素的生物利用度，从而解决高剂量氧化锌在断奶仔猪上的应用带来动物毒性、细菌抗性和环境污染等问题。
本发明的技术方案如下：
一种饲料级氧化锌的制备方法，采用碱式碳酸锌作为制备原料，制备温度250～300℃，将碱式碳酸锌在马弗炉中煅烧2h，制备得到饲料级氧化锌。
一种根据所述的制备方法得到的饲料级氧化锌，比表面积48.9m2/g，粒径10～30μ m，透射电镜分析得到的微观结构内部为由小梁支撑的中空结构，并且小梁含有丰富的孔腔。
所述的饲料级氧化锌，与市场上普通氧化锌、纳米氧化锌和多孔氧化锌相比，所述的饲料级氧化锌在人工胃液中不易被分解，对大肠杆菌K88、肠炎沙门氏菌CMCC5004和金黄色葡萄球菌ATCC25923具有更好的抗菌活性，对猪小肠上皮细胞IPEC-1具有更低的细胞毒性。
所述的饲料级氧化锌在断奶仔猪上的应用，断奶仔猪基础日粮添加低剂量300mg/kg饲料级氧化锌，用于降低断奶仔猪腹泻率。
所述的饲料级氧化锌与益生菌配伍在断奶仔猪上的应用，断奶仔猪基础日粮添加低剂量300mg/kg饲料级氧化锌和益生菌108CFU/kg丁酸梭菌ZJU-F1和109CFU/kg地衣芽孢杆菌用于降低断奶仔猪腹泻率，促进断奶仔猪生长，提高养分表观消化率，减少锌排放和肝脏沉积，改善断奶仔猪肠道屏障和免疫功能，提高肠道益生菌罗伊氏乳杆菌的丰度；所述的丁酸梭菌ZJU-F1(Clostridium butyricum)为现有菌株，中国微生物菌种保藏管理委员会普通微生物中心的保藏号为CGMCC No：8939；地衣芽孢杆菌(Bacillus licheniformis)购于浙江惠嘉生物科技股份有限公司。
本发明的有益效果：
该方法制备工艺简单，制备得到新型氧化锌比表面积48.9m2/g、粒径10～30μm。与市场上普通氧化锌、纳米氧化锌和多孔氧化锌相比，制备得到的新型氧化锌在人工胃液中不易被分解，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，对猪小肠上皮细胞IPEC-1具有更低的细胞毒性。低剂量新型氧化锌与高剂量的普通氧化锌、低剂量的纳米氧化锌和低剂量的多孔氧化锌相比，具有更好的降低断奶仔猪腹泻率的作用。将低剂量新型氧化锌和益生菌(丁酸梭菌和地衣芽孢杆菌)配伍可有效降低断奶仔猪腹泻率、促进断奶仔猪生长、提高养分表观消化率、减少锌排放，改善肠道屏障和免疫功能，提高肠道有益菌的丰度。
(1)300mg/kg新型氧化锌与益生菌配伍可以达到3000mg/kg高剂量普通氧化锌和抗生素配伍相同的防腹泻、促生长的效果。
(2)有效提高断奶仔猪干物质、粗蛋白和粗脂肪等养分表观消化率，降低锌在仔猪肝脏沉积，减少粪锌的排放。
(3)提高肠道菌群丰度以及有益生菌罗伊氏乳杆菌的丰度，改善肠道形态，提高肠道紧密连接蛋白，黏蛋白和抗菌肽表达，改善仔猪肠道屏障和免疫功能。
附图说明
图1是碱式碳酸锌和本发明制备的氧化锌透射电镜图。
图2是本发明制备的氧化锌在人工胃肠液中溶解度。
图3是本发明制备的氧化锌对大肠杆菌K88、肠炎沙门氏菌和金黄色葡萄球菌的抑制作用。
图4是本发明制备的氧化锌对猪小肠上皮细胞IPEC-1的毒性作用。
图5是本发明制备的氧化锌配伍益生菌对断奶仔猪肝脏和粪便中锌含量的影响。
图6是本发明制备的氧化锌配伍益生菌对断奶仔猪空肠形态的影响。
图7是本发明制备的氧化锌配伍益生菌对断奶仔猪肠道屏障和免疫功能的影响。
图8是本发明制备的氧化锌配伍益生菌对断奶仔猪肠道菌群的影响。
具体实施方式
以下结合附图和实施例对本发明做进一步的阐述。
实施例1
一种饲料级新型氧化锌的制备，具体制备方法如下：
采用碱式碳酸锌作为制备原料，使用热重分析仪测定碱式碳酸锌的热重分析曲线，在空气氛围中，加热升温速率20℃/min，加热温度范围50～500℃，在250～300℃有一个快速失重阶段，得到的适宜制备温度250～300℃。辅以适宜的通风、搅拌、升温和降温速率等条件，将碱式碳酸锌在马弗炉中煅烧2h，制备得到饲料级新型氧化锌，粒径10～30μm。
对制备得到的新型氧化锌做透射电镜分析，其微观结构如图1所示，其内部为由小梁支撑的中空结构，并且小梁含有丰富的孔腔。
表1.新型氧化锌比表面积和总孔体积
氧化锌活性与其比表面积正相关，通过统计不同形式氧化锌比表面积和总孔体积，结果如表1所示，新型氧化锌和与市场上的普通氧化锌、纳米氧化锌和多孔氧化锌相比，具有更大的比表面积(48.92m2/g)和总孔体积(0.57cm3/g)。
通过测定不同形式氧化锌在人工胃肠液中溶解度，由图2可知，与市场上的普通氧化锌、纳米氧化锌和多孔氧化锌相比，新型氧化锌在人工胃液中不易被分解，可使得更多的新型氧化锌以分子的形态到达肠道中发挥抗腹泻作用。人工胃肠液根据《2010中国药典(第二册)》中“附录XIX D二、体外药物释放度试验”进行配制。其中人工胃液(pH3.8)：0.1mol/L HCl溶液用1mol/L HCl溶液和1mol/L NaOH溶液调节pH至3.8；人工小肠液(pH6.8)：750ml 0.1mol/L HCl溶液与250ml 0.2mol/L Na3PO4溶液混合，用1mol/L HCl溶液和1mol/L NaOH溶液调节pH至6.8；人工结肠液(pH7.8)：750ml 0.1mol/L HCl溶液与250ml 0.2mol/L Na3PO4溶液混合，用1mol/L HCl溶液和1mol/L NaOH溶液调节pH至7.8。
由图3可知，500和1000mg/L新型氧化锌对大肠杆菌K88、肠炎沙门氏菌CMCC5004和金黄色葡萄球菌ATCC25923具有明显的抑制作用，其中200mg/L新型氧化锌对大肠杆菌K88具显著的抑菌效果。大肠杆菌K88是引起断奶仔猪腹泻的主要致病菌，新型氧化锌不仅可以使大肠杆菌K88缩短，而且还可以破坏其表面结构。
LDH释放率是通过检测细胞凋亡或坏死时，释放到胞外的乳酸脱氢酶，反映细胞膜的完整性，实现对细胞毒性的定量分析。由图4可知，与普通氧化锌、纳米氧化锌和多孔氧化锌相比，新型氧化锌具有最低的LDH释放率，提示新型氧化锌对猪小肠上皮细胞具有最低的细胞毒性。
综上所述，我们可得知，饲料级新型氧化锌制备工艺简单，同时具备高比表面积，在人工胃液中不易被分解，对大肠杆菌K88、肠炎沙门氏菌和金黄色葡萄球菌具有良好的抗菌活性以及对猪小肠上皮细胞IPEC-1细胞毒性低的优势。
实施例2
饲料级新型氧化锌在断奶仔猪上的应用，具体如下：
试验选取56头25日龄断奶仔猪(平均体重7.33kg，公母各半)分为4组，每组14头，试验为期14天。具体处理如下：基础日粮+3000mg/kg普通氧化锌组；基础日粮+300mg/kg纳米氧化锌组；基础日粮+300mg/kg多孔氧化锌组；基础日粮+300mg/kg新型氧化锌组。判断新型氧化锌对断奶仔猪生长性能和腹泻率的影响。
表2.新型氧化锌对断奶仔猪生长性能和腹泻的影响
由表2可知，300mg/kg低剂量饲料级新型氧化锌在断奶仔猪促生长上与3000mg/kg的高剂量普通氧化锌效果相当。与3000mg/kg普通氧化锌、300mg/kg纳米氧化锌和300mg/kg多孔氧化锌相比，断奶仔猪基础日粮添加300mg/kg新型氧化锌具有最低的腹泻率，提示低剂量的新型氧化锌可以作为高剂量普通氧化锌防治仔猪腹泻的有效替代品。
实施例3
饲料级新型氧化锌与益生菌配伍在断奶仔猪上的应用，具体如下：
试验选取90头25日龄断奶仔猪(平均7.88kg，公母各半)分为3组，每组3栏，每栏10 头，试验为期21天。具体处理如下：基础日粮+3000mg/kg普通氧化锌+抗生素组(“HOZO+Ant”组)；基础日粮+300mg/kg新型氧化锌+抗生素组(“LAZO+Ant”组)；基础日粮+300mg/kg新型氧化锌+益生菌组(“LAZO+Pro”组)。抗生素：100mg/kg喹乙醇、20mg/kg硫酸粘菌素和50mg/kg吉他霉素；益生菌：108CFU/kg丁酸梭菌ZJU-F1和109CFU/kg地衣芽孢杆菌。
表3.新型氧化锌配伍益生菌对断奶仔猪生长性能和腹泻的影响
由表3可知，300mg/kg低剂量新型氧化锌与益生菌配伍可基本达到3000mg/kg高剂量普通氧化锌和抗生素配伍的防腹泻、促生长的效果，避免了高剂量氧化锌带来的污染等问题
表4.新型氧化锌配伍益生菌对断奶仔猪养分表观消化率的影响
进一步测定新型氧化锌配伍益生菌对断奶仔猪养分表观消化率的影响，结果如表4所示，相比于3000mg/kg高剂量普通氧化锌和抗生素配伍，300mg/kg低剂量新型氧化锌配伍益生菌有效提高仔猪干物质、粗蛋白、粗脂肪、粗灰分以及磷的养分表观消化率。
由图5可知，低剂量新型氧化锌配伍益生菌能够有效减少断奶仔猪肝脏锌沉积，有效防止高剂量氧化锌导致断奶仔猪肝脏锌过载，同时有效减少了粪便中锌的排放对环境造成的污染。
由图6可知，低剂量新型氧化锌配伍益生菌能够避免高剂量普通氧化锌和抗生素配伍对断奶仔猪肠道造成的损伤，同时提高肠道微绒毛长度，增强肠道吸收面积。
由图7可知，低剂量新型氧化锌配伍益生菌能够提高断奶仔猪肠道紧密连接蛋白(Claudin-1和Occludin)、黏蛋白(MUC1、MUC2、MUC13、MUC20)和抗菌肽(pBD-1和pBD-2)的表达，从而改善断奶仔猪肠道屏障和免疫功能。
采用高通量测序对仔猪肠道微生物区系进行分析，结果如图8所示，低剂量新型氧化锌配伍益生菌提高断奶仔猪肠道菌群丰度以及益生菌罗伊氏乳杆菌的丰度。罗伊氏乳杆菌能 够促进动物肠道消化吸收，抑制肠道内致病菌的增殖。
综上所述，我们可得知，300mg/kg低剂量饲料级新型氧化锌配伍益生菌可以达到与3000mg/kg高剂量普通氧化锌配伍抗生素相同的防腹泻、促生长效果。同时提高有效提高断奶仔猪养分表观消化率，降低锌在仔猪肝脏沉积，减少粪锌的排放，改善肠道形态、肠道屏障和免疫功能，提高肠道菌群丰度以及有益菌罗伊氏乳杆菌丰度。
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对发明范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明的保护范围应以所附权利要求为准。

Claims (5)

  1. 一种饲料级氧化锌的制备方法，其特征在于，采用碱式碳酸锌作为制备原料，制备温度250～300℃，将碱式碳酸锌在马弗炉中煅烧2h，制备得到饲料级氧化锌，比表面积48.9m2/g，粒径10～30μm。
  2. 一种根据权利要求1所述的制备方法得到的饲料级氧化锌，其特征在于，比表面积48.9m2/g，粒径10～30μm，透射电镜分析得到的微观结构内部为由小梁支撑的中空结构，并且小梁含有丰富的孔腔。
  3. 根据权利要求2所述的饲料级氧化锌，其特征在于，与市场上普通氧化锌、纳米氧化锌和多孔氧化锌相比，所述的饲料级氧化锌在人工胃液中不易被分解，对大肠杆菌K88、肠炎沙门氏菌CMCC5004和金黄色葡萄球菌ATCC25923具有更好的抗菌活性，对猪小肠上皮细胞IPEC-1具有更低的细胞毒性。
  4. 一种根据权利要求2所述的饲料级氧化锌在断奶仔猪上的应用，其特征在于，断奶仔猪基础日粮添加低剂量300mg/kg饲料级氧化锌，用于降低断奶仔猪腹泻率。
  5. 一种根据权利要求2所述的饲料级氧化锌与益生菌配伍在断奶仔猪上的应用，其特征在于，断奶仔猪基础日粮添加低剂量300mg/kg饲料级氧化锌和益生菌108CFU/kg丁酸梭菌ZJU-F1和109CFU/kg地衣芽孢杆菌用于降低断奶仔猪腹泻率，促进断奶仔猪生长，提高养分表观消化率，减少锌排放和肝脏沉积，改善断奶仔猪肠道屏障和免疫功能，提高肠道益生菌罗伊氏乳杆菌的丰度；所述的丁酸梭菌ZJU-F1(Clostridium butyricum)为现有菌株，中国微生物菌种保藏管理委员会普通微生物中心的保藏号为CGMCC No：8939；地衣芽孢杆菌(Bacillus licheniformis)购于浙江惠嘉生物科技股份有限公司。
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