猪用抗生素的替代.ppt
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1、Alternatives of antibiotics for pigs 猪用抗生素的替代,Defa Li 李德发,Ministry of Agriculture Feed Industry Centre 农业部饲料工业中心,2006年11月30日于北京,猪营养,营养需要 与代谢,分子生物学,Who operates MAFIC Research,植物提取物,免疫学,微生物学,分析化学,营养与免疫,基因工程菌,细胞生物学,分子营养 与肉品质,团队研究,Antibiotics 抗生素,Residue 残留,Resistance 耐药性,Harmful to animal and human he
2、alth 影响动物和人类的健康,Reduce the use of antibiotics 减少抗生素的使用,EU 欧盟,China 中国,Current situation and future development 现状与展望,Alternatives of antibiotics 抗生素替代,Antimicrobial peptides 抗菌肽,Herbal extracts 天然提取物,Enzyme preparations 酶制剂,Probiotics 益生素,Probiotics 益生素,Lactobacillus 乳酸杆菌,Mechanism 机理研究,Application
3、应用研究,绿色荧光蛋白 (GFP)标记,蛋白组学,粘附位点,无抗发酵饲料,蛋白差异,复合发酵培养,悉生动物模型,信号传导,作用机制,Targeting distribution 目的细菌在肠道中的定位分布情况,其与肠道菌系之 间的关系 Specific passway and function 具体作用途径与功能的研究,Experimental model of gnotobiotic animals 悉生动物实验模型,悉生动物(已知菌动物),空白组,益生菌,致病菌,益生菌 + 致病菌,Development of antibiotics-free fermented feed 无抗发酵饲料的
4、开发,Fermentative respiratory membrane 发酵呼吸膜,Single way valve 单向阀门,CO2 venting, preventing air to enter 排出袋内微生物产生的CO2,阻止大气进入,A, E. coli K88 大肠杆菌K88 B, E. coli K99 大肠杆菌K99 C, S. typhimurium 鼠伤寒沙门氏菌 D, Staph. Aureus 金黄色葡萄球菌,Int. J. Gener. Mol. Microbiol., X. H. Guo, Defa Li, 2006,Inhibitory activity of
5、Bacillus spp. 139 against pathogenic bacteria, but not Lactobacillus 外源芽孢杆菌可抑制有害菌,对乳酸菌无影响,Multi-strains combination, creating anaerobic condition automatically 多菌种组合,自身创造厌氧环境,Yeast 酵母菌,Bacillus spp. 芽胞杆菌,好氧,兼性,兼性,Lactobacillus reproduction in anaerobic condition 乳酸杆菌厌氧发酵,Inoculating, packing, then f
6、ermentation 先接种、包装,后发酵,发酵,轻工业副产物:玉米浆、果渣,Effect of fermentation time on energy and gossypol 能量与游离棉酚随发酵时间的变化,Decrease the content of gossypol 降低游离棉酚的含量,Effects of antibiotics-free fermented feed on growth performance in pigs 无抗发酵饲料对猪生产性能的影响,P 0.05,RFID system RFID 追溯系统,基因芯片,Gene chip :The on line exam
7、ination technique of antibiotics 基因芯片:快速检测分割肉中的抗生素含量(20-100 ppb),Proteomics 蛋白质组学,Proteomics analysis of interaction between Lactobacillus and intestine 乳酸杆菌和肠道细胞相互作用的蛋白质组学研究,J. Nutr., J. J. Wang, Defa Li, 2006,蛋白质组学,双向电泳,样品制备,数据库比对鉴定蛋白,相互作用机制,生物功能,差异蛋白,质谱分析,图像分析,胶内酶切,蛋白,Lactobacillus 乳酸杆菌,Intestina
8、l epithelial cells 肠道上皮细胞,等点聚焦,SDS-PAGE,Experimental design 试验设计,Intestinal cells,Adhered or not,Lactobacillus fermentum I5007 cells,Culture medium, 37C 37C温箱,4 h,Rabbit jejunum 兔子空肠,Two-dimensional gel electrophoresis 双向电泳,37C 1 h,Mass spectrometry 质谱,Differential proteins 差异蛋白,Lactobacillus in rab
9、bit jejunum culture 兔子空肠培养条件下乳酸杆菌表达差异的蛋白,Electrophoresis, F. Yang, Defa Li, 2006,Fructose-6-phosphate Phosphoketolase (F6PPK) 6-磷酸果糖磷酸酮酶,Lactate dehydrogenase 乳酸脱氢酶,Dihydrolipomide dehydrogenase 二氢硫辛酸脱氢酶 (丙酮酸脱氢酶系),糖酵解,三羧酸循环,磷酸戊糖途径,减少氧自 由基产生,NADPH 生成增加,有利于乳酸杆菌存活,Intestinal cells after Lactobacillus a
10、dhesion 肠细胞被乳酸杆菌粘附条件下表达差异的蛋白,Electrophoresis, F. Yang, Defa Li, 2006,Voltage-dependent anion channel 1 电压依从性阴离子通道1,Glutathione transferase 谷胱甘肽转移酶,Heat shock protein gp96 热应激蛋白gp96,调控膜通透性,改善机体免疫,减少氧化应激,改善肠道完整性 及其健康,Synbiotics 合生素,Functional mechanism of synbiotics 合生素的作用机理,Synbiotics 合生素,Bacillus su
11、btilis 枯草芽孢杆菌,Astragalus, aloe polysaccharide, etc. 黄芪多糖,芦荟多糖等,Immunity 免疫力,Antibacterial substance 抑菌物质,Organic acid 有机酸,Health 动物健康,Bacteriophage 产生抗菌素 挥发性脂肪酸:乙酸、丙酸、丁酸 Tolerance of the intestinal environment 耐受肠道内环境,Bacillus subtilis 枯草芽孢杆菌,ZnO 氧化锌,Hypothesis 假设,Intestine function 肠道功能,Enteric ner
12、vous system 肠神经系统,Feed intake 采食量,Diarrhea 腹泻,生产性能 Growth performance,Gastrointestinal hormones 胃肠激素,Mast cell mediators 肥大细胞介质,Intestinal IGF-1 system 小肠IGF-1系统,Hormone on eating 采食相关激素,ZnO,Effects of dietary supplement of 3000 mg/kg ZnO on the expression of fundic gland Ghrelin gene in piglets 日粮中
13、添加3000 mg/kg氧化锌对断奶仔猪胃底腺Ghrelin基因表达的影响,P = 0.008,Increase the expression of the Ghrelin gene 显著提高断奶仔猪胃底腺Ghrelin基因的表达, 提高采食量,Improve the intestinal function 药理剂量氧化锌促进仔猪肠道健康,The small intestinal morphology of weaned pigs receiving a diet with or without supplementation of 3000 mg/kg Zn 日粮中添加3000 mg/kg氧
14、化锌ZnO对仔猪小肠形态 的影响,J. Nutr., X. L. Li, Defa Li, 2006,Increase the expression of IGF-1 gene 提高小肠 IGF-1 基因的表达 通过IGF-1的旁分泌作用促进仔猪肠道健康和生长,Effects of dietary supplement of 3000 mg/kg ZnO on the expression of small-intestinal mucosa IGF-1 gene in piglets 日粮中添加3000 mg/kg氧化锌对仔猪小肠粘膜IGF-1基因表达的影响,J. Nutr., X. L.
15、Li, Defa Li, 2006,Histamine concentrations in the small intestine of weanling piglets fed diets containing normal and high-levels of ZnO 高锌日粮对断奶仔猪小肠组胺浓度的影响,Decrease the release of the histamine in the small intestine 高锌显著降低断奶仔猪小肠组胺的释放 稳定小肠通透性,The numbers of mast cells (cell/cm2) in the small intesti
16、ne of weanling piglets fed diets containing normal or high-levels of zinc oxide 高锌日粮对断奶仔猪小肠肥大细胞数量的影响,Decrease the numbers of the mast cells in the small intestine 高锌显著降低断奶仔猪小肠各层肥大细胞数 改善小肠功能,mRNA and protein levels for the stem cell factor (SCF) gene in the mid jejunum of weanling piglets fed diets c
17、ontaining normal and high-levels of ZnO 高锌对空肠中段干细胞因子表达(基因与蛋白)的影响,Decrease the expression of SCF 高锌显著降低空肠干细胞因子的表达 调控肥大细胞数量及介质释放,P = 0.016,Protein,Normal ZnO,High ZnO,P = 0.024,mRNA,Normal ZnO,High ZnO,The effects of dietary zinc on the changes in Isc (IscZn) and in G (GZn) 日粮锌对短路电流和电导率改变的影响,Comp. Bio
18、chem. Physiol., Z. K. Feng, 2006,Zn improves tight junction permeability 高锌降低断奶仔猪小肠粘膜跨上皮电位差 维持肠道紧密连接的完整性,Enzyme preparations 酶制剂,Complex enzyme-mutation and selection 复合酶诱变选育,Aspergillus sulphureus-MAFIC001 可高产复合酶的硫色曲霉及其诱变筛育方法和固态发酵制备饲用复合酶的应用,High yield of complex enzyme: xylanase, -glucanase, pectin
19、ase, etc. 高产复合酶:木聚糖酶、 -葡聚糖酶、果胶酶等 Industrial production 工业化生产,商业利润超过1000万元,Aspergillus niger-MAFIC005 饲用非淀粉多糖酶高产菌种的选育及其固态发酵生产技术,High yield of complex enzyme: xylanase, -glucanase, pectinase, etc. 高产复合酶:木聚糖酶、葡聚糖酶、果胶酶等,Existing questions and the methods to increase the yield of -mannanase 存在问题及提高-甘露聚糖酶
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