不用抗生素条件下布拉迪酵母改善动物肠道的应用

,Use of S. Boulardii to improve Gut health under antibiotics free trend,Pierre Lebreton, April 2014,Pierre LEBRETONIng. Agriculture, Pig nutrition specialistTechnical Director of Tecnor-Sofac FranceSpecialised in sows and piglets feeds production,Consultant for feed industries and pig farms,Consultant for Lallemand companiePreviously part of the french Provimi pig team法国资深猪饲料配方和生产顾问，从业16年曾担任法国普乐维美猪饲料总配方师、法国Brittany养猪联合会技术顾问、现任法国Tecnor-Sofac公司技术总监，同时被法国养猪联合会和拉曼动物营养聘请问技术顾问。,Global conditionning factors for piglets abilities:制约仔猪生产性能的因素,Piglet weight, maturity, vitality at birth仔猪出生重、出生时的成熟度与活力,Colostrum intake, Colostrum quality初乳的摄取量、初乳的品质,Prestarter intake, weaning prparation教槽料/过渡料的采食、断奶准备,Housing conditions, contamination from sow房舍条件、母猪病原菌的垂直传递,Milk production, milk quality猪奶的供给量、猪奶的品质,21,2.a,Contamination from the sow母猪病原菌的垂直传递,The quality of the faeces influence the composition of the piglets gutFiber quality and content of the feed (gestation &lactation) 母猪粪便的优劣会影响仔猪消化道微生物的组成-纤维的品质和饲粮成分（妊娠料和泌乳料）决定母猪粪便的优劣,Weaning : Stresses for the piglet断奶：仔猪遭受应激,Stop in the maternel link强制与母猪分离,Change of environnement环境的变化,Modification of social organisation分群使社交模式改变,Modification of feeding and feeds饲喂制度和饲粮的改变,Immunitary weakness免疫系统尚不成熟,STRESSUnder-NUTRITION,Immature Digestif System尚不成熟的消化系统,Main consequences : 主要不良后果 Microbes and virus are widely exchanged 病原微生物和病毒混杂- environment 环境- stress 应激- decrease of immunity 免疫力降低- stress on the microflora 应激导致肠道菌群变化 Frequent pathological events 疾病频发 mainly enteric aspects (diarrhoeas, dema.) respiratory 主要是肠道疾病（腹泻、水肿病）和呼吸道疾病 symptoms, mortality 有临床症状且死亡率高 - feed efficiency decreased (ADG,FCR) 饲料转化率下降,.,Before Weaning,After W,incredible number of new species discovered:被发现的新微生物菌种数量庞大 854 different bacteriasidentified in the feces of an unweaned piglet一头哺乳小猪粪中鉴定出854种不同微生物Lactobacilli very limited counts!乳酸杆菌的比例非常小,Stress of the microflora : Deep sequencing method微生物菌群的应激：深度测序技术,断奶前,.,After Weaning,Big changes appears!断奶导致肠道菌群发生重大改变Cellulolitic bacterias are very sensitive to stresses纤维分解菌对应激非常敏感,Stress of the microflora : Deep sequencing method微生物菌群的应激：深度测序技术,断奶后,Stress of the microflora : role of feed intake微生物菌群的应激：采食量的作用,Week of lactation泌乳期,Days after weaning断奶后几天内的变化,Age, weeks56周龄之后,Average consumption of dry matter平均干物质摄入量,断奶Weaning,Weaning : Stresses for the GIT断奶：对胃肠道的影响,群体和日粮的改变,采食量降低,肠道的营养缺乏,粘膜功能受损,消化不良吸收障碍,生长表现不佳腹泻感染,炎症反应,细菌抗原和毒素摄入增加菌体位移增加,Microscopy and histomorphology of the villi before and after weaning小肠绒毛在断奶前后镜检和组织形态学的改变 Dr John Pluske Murdoch Uty,Impact on the villis quality :对小肠绒毛质量的影响,Gut structure in the small intestine 小肠结构,Before weaning断奶前,After weaning断奶后,After weaning断奶后,Before weaning断奶前,12,Gut is the largest immune organ肠道是最大的“免疫”器官,Huge amounts and diversity of antigens due to food and gut microbes摄入的食物和肠道微生物形成动物肠道内数量庞大且多样化的抗原物质All components of the defense system: physical barrier, mucus, specialised immune cells防御体系齐全：物理性屏障、粘膜免疫、特殊的免疫细胞Dense network of nerves and blood vessels: connected神经与血管网络分布密集且紧密相联,Effects on immunity 对免疫的影响,Interaction between bacteria, intestinal cells and dendritic cells细菌、肠道细胞、树突细胞的相互作用,MAMPs: microorganism-associated molecular patterns; PRRs: pattern recognition receptors,Lebeer et al., 2010, Nature Reviews Microbiology 8,The Ig concentration from the colostrum declines very rapidly ; the piglets own immune system progressively starts producing its own antibodies (25) 初乳中的免疫球蛋白浓度迅速下降；仔猪自身免疫系统逐渐建立产生自体抗体,Ig in the serum (mg/ml)血清中免疫球蛋白水平,Piglet Ig endogenous synthesis 仔猪免疫球蛋白内源合成量,Ig from the Colostrum初乳中的免疫球蛋白,Total Ig level 总抗体水平,Adult Ig level 成年免疫球蛋白水平,Birth,J.28,J.21,J.14,J.7,Days,Immunity , first defensive line: 免疫, 第一道防线,(25) Euzeby, 1989, Bull Acad Natl Med,Lower protection: higher susceptibility to pathogens抗体保护较低：对病原菌易感性较高,Total Immunoglobulines level along the time随着时间延长仔猪体内总免疫球蛋白水平的变化,Effects of weaning on enz. activity (pancras)断奶对消化酶活性的影响 （胰腺）,Trypsine胰蛋白酶,Chymotrypsine胰凝乳蛋白酶,Lipase酯酶,Amylase淀粉酶,Days after weaning断奶后日龄,(*),(*),(*) : enzymatic activities in relation to weaning, weaning = 100相对于断奶时水平的消化酶活性，断奶酶活设为100,Days after weaning断奶后日龄,Suckling piglets 哺乳小猪24 day old weaned Piglets but sick and kept untreated 24日龄断奶仔猪、疾病当尚未给药治疗31 day old weaned and healthy Piglets, 7 days post-weaning 31日龄断奶且健康仔猪，断奶后7天的情况 (having received Zinc oxide and Antibiotics) 已经摄入氧化锌和抗生素Abbreviations: ApA aminopeptidase A 氨基肽酶A ; ApN aminopeptidase N 氨基肽酶N ; DPP IV dipeptidilpeptidase IV 二酰肽肽酶IV.,Thymann et al.,Specific enzymatic activities in the upper gut and the pancreas上消化道和胰腺中的特定酶的酶活水平,Poor digestion lot of undigested substrates available for bacterial growth (protein!)消化不良大量未消化食糜成为病原微生物生长的营养底物,Effects of weaning on enz. Activity断奶对消化酶活性的影响 （胰腺）,Main Solutions used to avoid antibiotics :避免使用抗生素的主要方案,Impact of prestarter intake, 教槽/过渡料采食量的影响Impact of feed quality before and after weaning,断奶前后饲料品质的影响Help of Levucell SB布拉迪的应用,这里是否改成益生菌类产品的应用提及欧洲市场上的乳酸杆菌、粪链球菌、酵母菌、布拉氏酵母最后说他更倾向于使用布拉迪的原因,Objective from :关注目标5 days before weaning to 5 days after weaning断奶前5天至断奶后5天 Feed and protect the gut as an immune organ before looking for feeding the piglet for growth首先营养和保护消化道使其充分发挥成熟免疫器官的功能其次才是饲养仔猪促进其生长,First : Feed consumption首先： 饲料采食量,First : Feed consumption首先： 饲料采食量,Week of lactation,Days after weaning,Age, weeks,Average consumption of dry matter平均干物质消耗量,10 days,断奶weaning,Week of lactation泌乳期,Days after weaning断奶后几天内的变化,Age, weeks56周龄之后,Trials 2013, Tecnor-sofac : impact of pre starter consumption,Exemple of trials made last year on 3 commercial farms, 3 batches, 100% of piglets, Tecnor-Sofac Feeds2013年3个商业猪场连续3个批次仔猪的饲养成绩，全场使用Tecnor-Sofac的饲料,Farm N1,Farm N2,Farm N3,+42%,+82%,Quantiti consummed (g),Quantiti consummed (g),3 Batches,Total quantity of feed per litter,Total quantity of feed per litter,1st days after weaning,Total quantity per piglet,Total quantity per piglet,每窝饲料消耗量,断奶后一天的仔猪采食量,40% of ADG variation from de 0 to 34 days is explained by feed intake in the first week after weaning34日龄前平均日增重的40%取决于断奶后第一周的采食量,Bruininx 2002,First : Feed consumption,Protin from milkLactose乳制品提供蛋白和乳糖,Equilibrium between raw and cooked crals:Wheat, barley, biscuits熟化与未加工谷物的平衡小麦、大麦、饼干,Adapted Nutritional levels适合的营养水平 ,Organic Acids有机酸,Enzymes消化酶,Levucell SB布拉迪,Fibers with high Hydratation capacity高水合能力的纤维,PigletsFeeds,Fatty acids and fat脂肪酸和脂肪,AntiOxydants, Vit抗氧化剂与生物素,Methyl donnors甲基供体,Essencial oils精油,.,Second : Feed Quality 其次： 饲料的品质,Second : Feed Quality =,More than nutritionnal levels,Raw material quality 饲料原料的品质= Protein quality (high sensibility) 蛋白的品质（较高的敏感性） Fiber quality (soluble & insoluble) 纤维的品质（可溶&不可溶） Fat quality 脂肪的品质 Starch quality 淀粉的品质 Milk products quality 乳制品的品质 Probiotics 益生菌的选择 . Mycotoxins 霉菌毒素 Oxydations 氧化,Slection of safety raw materials安全饲料原料的选择,Limit soja meal 限制豆粕的使用Technological Process, antinutritionnals factors and take out their sensibilising factors ( inflammation)适宜的加工处理技术能降低抗营养因子水平、清除特异性过敏原，降低炎性反应发生风险,Farine soya standard,Conc. protines soya,Slection of safety raw materials安全饲料原料的选择,Soja meal 豆粕,Soja meal treated 熟化豆粕,Plasma 血浆,Milk serum 乳清,Soja meal treated 熟化豆粕,Patatoe protein 土豆蛋白,Synthetic amino acids 合成氨基酸,Exemple of Protein composition in special formulation (21 day weaning) : 21日龄断奶典型配方的蛋白组成,Adapted starchs : 适宜的淀粉,Trial Wheat Cooked wheat,Cereals variety and quality is important :Use of cooked wheat and oat, cakes and BarleyCooked Rice could be very interesting谷物多样性和品质均至关重要使用熟化的小麦、燕麦、饼干和大麦熟化大米的效果可能也非常好,Exemple of T-S Trials,Adapted starchs : 适宜的淀粉,Exemple of starch content in special formulation (21 day weaning) :21日龄断奶配方中淀粉来源的举例,Cake 蛋糕,Barley大麦,CookedWheat熟化小麦,High Quality Wheat高品质小麦,Adaptated Feeds :适宜的饲料,In my company we developed 9 different types of starters : 9种类型的淀粉针对不同的应用方案,Special gut protection, (days before weaning, 5 days after), 针对保护肠道，断奶前至断奶后5天使用Spcial diahrea sensibility, low protein level Spcial diahrea sensibility, normal protein level 针对腹泻敏感性选择淀粉，并配合不同蛋白水平Spcial weaning 19 days, 针对不同断奶日龄选择淀粉Spcial weaning 23 days, 并考虑是否需要着重保护菌群稳定性Spcial weaning 28 days , with flora protectionSpcial weaning 28 days without high protectionSpcial weaning 23 days, with antibiotic use (probiotic inside)Spcial weaning 28 days, with antibiotic use (probiotic inside)针对不同断奶日龄且使用抗生素的情况，此时必须添加益生菌保护,Exemple of extreme formulation,Spcific feed free antibiotic trend :,Raw materials,Exemple of nutrient levels :,Exemple of nutrient levels :,Amino acids :,Exemple of nutrient levels :,Exemple of premix content :,Extract,Adaptated solutions : exemple of 21 days weaning product21日龄断奶的方案实例,Acidification 酸化剂Sorbic + Benzoic +.山梨酸+苯甲酸+,Essentials oils 精油Thymol/Eugenol/Carvacrol麝香草酚/丁香油酚/香芹酚,Resistant Starch抗性淀粉,Fat emulsifier乳化剂,Clay : smectite蒙脱石,Probiotic益生菌,.,Soluble&Insoluble fiber可溶和不可溶纤维,Good Flora,Adaptated Probiotic : 适宜的益生菌,Several probiotics have been tested, selection has been done based on :根据下面的选择依据已经对许多益生菌进行过筛选1- resistance again antibiotics, 抗生素敏感性2- resistance again acidification, 酸化剂敏感性3- ability to promote cellulolitic flora, 纤维分解菌促进能力4- ability to promote lactic flora, 乳酸菌的促进能力5- direct property to struggle again E. Coli 直接抵抗大肠杆菌的能力6- direct effect on inflamation 对炎症反应的直接效果7- heat resistance during processing 饲料加工工艺的热稳定性,Live yeast preferred 活酵母最终胜出,4,1,LEVUCELLSB :SPECIFIC PROBIOTIC YEAST布拉迪特殊的益生菌酵母,Identity and Selection 鉴定与筛选b) Strain : Saccharomyces cerevisiae boulardii CNCM I-1079菌株：酿酒酵母布拉氏亚种CNCM I-1079菌株c) LEVUCELL SB : Mode of Action 作用模式d) LEVUCELL SB : Main Applications and Role 主要应用与效果,Third : 第三点,5,1.a,: A WORLD LEADER IN YEAST AND BACTERIA PRODUCTION世界顶尖的酵母和细菌生产供应商,1. Global leader in microbial selection and screening: More than 1,000 yeast strains in Lallemand culture collection.拉曼拥有上千株菌株的微生物种质资源2. State of the art production processes for microbial products: More than 25 specialized yeast & bacteria production plants around the world. 全球超过25家专业化的酵母和细菌发酵工厂3. Effective animal research and biotechnology network: 3 labs fully dedicated to screening, processing and stability, 三所实验室致力于菌株筛选、加工和稳定性 Research partnerships with world-class expertise centres: University of Nottingham, INRA, BRI 世界顶尖的研究机构长期合作研究诺丁汉大学、法国农业科学院、加拿大国家生物技术研究所.,Identity and Selection 鉴定与筛选,1.a, Bakers yeast : Maltose utilization Brewers yeast : Bitter Attenuation Wine yeast : Flavor, Color Distillers yeast: Fermentation rate, alcohol tolerance,Objectives Regulation of intestinal microflora 协调肠道菌群 Stimulation of immune system 刺激免疫系统 Improved intestinal structure and function 改善肠道结构与功能,6,Identity and Selection 鉴定与筛选,7,1.b,OLD STORY BUT PROMISING FUTURE!,Saccharomyces cerevisiae boulardii 酿酒酵母布拉氏亚种,1920. 发现 Identification of the active ingredient of a traditional Asian medicine (infusion of lychee hulls) against infant diarrhea : Dr Boulard discovered and isolated a specific yeast strain and named it boulardii.1962. 治疗人腹泻的药物上市 First form of boulardii active yeast recognized and marketed as a probiotic in human medicine. Since then, a huge amount of fundamental research on mode of action and application of boulardii in human medicine has been generated.1993. 欧盟首次批准用于动物 First EU registration of a S.c.boulardii (CNCM I-1079 / LEVUCELL SB) specific active yeast for use as a gut flora stabilizer in animal nutrition.1997. 完成分类学鉴定boulardii was classified as a variety of Saccharomyces cerevisiae. Its genetic similarity and its functional differences are recognized.Today, 广泛使用LEVUCELL SB is authorized and marketed in more than 44 countries.,Research bibliography booklet : Selection of 48 references related to S.c.boulardii in swine feeding. Abstracts in their original version.,8,1.b,S.c.boulardii is one of the most documented probiotic strain. The product already has an important scientific background and is still under scope for further documenting new applications and better understanding of its specific mode of action. LEVUCELL SB matches perfectly the trend to develop sustainable, effective and documented solutions for animal production.,Saccharomyces cerevisiae boulardii酿酒酵母布拉氏亚种,这页可以去掉,42,LEVUCELLSB : Mode of Action,Objective : IMPROVEMENT OF GUT FUNCTION AND NATURAL SELF-DEFENSES改善肠道天然自体防御体系,1.c,LEVUCELLSB : Mode of Action 作用模式,10,1.c,1st LINE OF DEFENSE :REGULATION OF INTESTINAL MICROFLORA第1道防线: 调整肠道微生物菌群,Stimulation of positive microflora刺激有益菌增殖,Reduced attachment of undesirable bacteria减少病原菌附着,Oxygen consumption清除残余氧,Adhesion粘附作用,Neutralisation中和作用,LEVUCELLSB : Mode of Action,Unique mode of action Neutralisation of microbial toxins (C.difficile, E.coli, .)特异性细菌毒素中和功能,48,Faster O2 consumption with LEVUCELLSB than with the other yeast strains布拉迪 比其他酵母菌株的耗氧速度更快,LEVUCELL SB role in the hindgut Oxygen scavenging in the caecum and the colon.盲结肠清除残余氧 Better anaerobic conditions. 提高厌氧微环境 Better fermentation rate. 提高发酵速度 More energy released from the same diet shorter time (SCFA energy)相同日粮因发酵产生短链脂肪酸供能而提供更高能值,Oxygen consumption,10 mL of a 0.1% resazurin solution+990 mL distilled water+5 g NaHCO3,16,1.c,2nd LINE OF DEFENSE :MAINTENANCE OF GOOD INTESTINAL STRUCTURE AND FUNCTION第二道防线：维持肠道正常结构和功能,Improved villus heightand crypt depth增加绒毛长度、隐窝深度,Improved barrier function.维持肠道上皮天然屏障功能,(University of Nottingham 2012),(University of Milan 2002),LEVUCELLSB : Mode of Action,LEVUCELLSB : Mode of Action,46,Improvement of physical barrier (gut)肠道物理屏障的改善,P 0,05,P 0,01,P 0,01,P 0,05,Control,+Levucell SB,Tight junction 紧密连接,47,跨膜蛋白,长度约50400nm，无缝隙；跨膜结构（紧密连接封闭索）由跨膜蛋白+胞质附着蛋白（粘附分子）构成。主要跨膜蛋白为封闭蛋白Claudin和密封蛋白Occludin。ZO具有封闭和维持栅栏栅：阻止2个不同功能区之间相互弥散形成细胞极性屏蔽：对分子大小、离子类型、细胞渗透性有选择性，调节胞间小分子运输,跨膜蛋白,紧密连接粘附因子,胞质附着蛋白ZO,肌球蛋白,肌动蛋白,Piglets fed LEVUCELLSB show up-regulated genesin the colon 饲喂布拉迪的仔猪呈现结肠上调基因,Watts et al., unpublished. MCOE,48,4 / 10related to gut wall function,免疫与神经内分泌,催产素信号,调节肌动蛋白稳定细胞骨架,紧密连接,信号转换,信号转换,细胞骨架构型,12,1.c,3rd LINE OF DEFENSE :ANTI-INFLAMMATORY AND IMMUNE PROPERTIES第三道防线：抗炎性和免疫特性,Less piglets mortality following a LPS endotoxin challenge脂多糖内毒素攻毒后仔猪死亡率降低,Less inflammation: decreased serum concentration of IL-1 (pro-inflammatory cytokine)降低炎性反应减少血清中促炎性白介素浓度,LEVUCELLSB : Mode of Action,Collier et al. 2010.,50,LEVUCELLSB improves pathogen recognition capacityin weaned piglets (1 week post weaning),Le Bon et al. unpublished MCOE,More toll-like receptors = beacons recognizing groups of micro-organisms,2009: Nobel Prize for TLR discovery = cutting edge research,ileum,Le Bon et al. unpublished MCOE,51,More toll-like receptors = beacons recognizing groups of micro-organismsToll 受体蛋白 (TLRs),2009: No