不用抗生素条件下布拉迪酵母改善动物肠道应用课件.ppt

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1、UseofS.BoulardiitoimproveGuthealthunderantibioticsfreetrendPierreLebreton,April2014Pierre LEBRETONIng. Agriculture, Pig nutrition specialistTechnical Director of Tecnor-Sofac FranceSpecialised in sows and piglets feeds productionConsultant for feed industries and pig farms,Consultant for Lallemand c

2、ompaniePreviously 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制约仔猪生产性能的因素PPiglet weight, maturity, vitality at birth仔猪出生重、出生时的成熟度与活力PColostrum

3、 intake, Colostrum quality初乳的摄取量、初乳的品质PPrestarter intake, weaning prparation教槽料教槽料/过渡料的采食过渡料的采食、断奶准备、断奶准备PHousing conditions, contamination from sow房舍条件、母猪病原菌的垂直传递PMilk production, milk quality猪奶的供给量、猪奶的品质42.aContamination from the sow母猪病原菌的垂直传递母猪病原菌的垂直传递The quality of the faeces influence the compo

4、sition of the piglets gut-Fiber 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Modifica

5、tion 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-NUTRITIONImmature 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应激导致肠道菌群变化 F

6、requent pathological events 疾病频发疾病频发 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 WeaningAfter W- incredible number of new species discovered:被发现的新微生物菌种数量庞大 - 854 different bacteriasid

7、entified 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：深度测序技术微生物菌群的应激：深度测序技术断奶前.After WeaningBig 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

8、常敏感Stress of the microflora : Deep sequencing method微生物菌群的应激：深度测序技术微生物菌群的应激：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WeaningWeaning : Stresses for th

9、e GIT断奶：对胃肠道的影响断奶：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小肠绒毛在断奶前后镜检和组织形态学的改变 Dr John Pluske Murdoch UtyImpact on the villis quality :对小肠绒毛质量的影响对小肠绒毛质量的影响 Gut structure in the sm

10、all intestine 小肠结构小肠结构 Before weaning断奶前断奶前After weaning断奶后断奶后After weaning断奶后断奶后Before weaning断奶前断奶前12Gut is the largest immune organ肠道是最大的肠道是最大的“免疫免疫”器官器官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

11、 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细菌、肠道细胞、树突细胞的相互作用MAMPs: microorganism-associated molecula

12、r patterns; PRRs: pattern recognition receptors Lebeer et al., 2010, Nature Reviews Microbiology 8The 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

13、/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BirthJ.28J.21J.14J.7DaysImmunity , first defensive line: 免疫免疫, 第一道防线第一道防线(25) Euzeby, 1989, Bull Acad Natl Med Lower protection: higher susceptibility to pathogens抗体保护较

14、低：对病原菌易感性较高抗体保护较低：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断奶对消化酶活性的影响 （胰腺）（胰腺）Trypsine胰蛋白酶胰蛋白酶Chymotrypsine胰凝乳蛋白酶胰凝乳蛋白酶Lipase酯酶酯酶Amylase淀粉酶淀粉酶Days after weaning断奶后日龄断奶后日龄(*)(*)(*) : enzymat

15、ic activities in relation to weaning, weaning = 100相对于断奶时水平的消化酶活性，断奶酶活设为相对于断奶时水平的消化酶活性，断奶酶活设为100Days after weaning断奶后日龄断奶后日龄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 (ha

16、ving 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 substrat

17、es 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断奶对消化酶活性的影响 （胰腺（胰腺） Main Solutions used to avoid antibiotics :避免使用抗生素的主要方案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

18、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断奶前断奶前5天至断奶后天至断奶后5天天 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

19、长First : Feed consumption首先：首先： 饲料采食量饲料采食量 First : Feed consumption首先：首先： 饲料采食量饲料采食量 Week of lactationDays after weaningAge, weeksAverage consumption of dry matter平均干物质消耗量平均干物质消耗量10 days断奶weaningWeek of lactation泌乳期Days after weaning断奶后几天内的变化Age, weeks56周龄之后2 BatchesDuring lactation*+ 8 %*48h aprs s

20、evrage+ 14 %During lactation*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 N1Farm N2Farm N3Total quantity of feed per litterQuantiti

21、consummed (g)*3 BatchesDuring lactation 泌乳期泌乳期+150%Total quantity per piglet *1st days after weaning+62%+42%+82%Quantiti consummed (g)Quantiti consummed (g)3 BatchesTotal quantity of feed per litterTotal quantity of feed per litter1st days after weaningTotal quantity per piglet Total quantity per pi

22、glet 每窝饲料消耗量断奶后一天的仔猪采食量 Weaning with Pr-starter 使用教保料断奶使用教保料断奶 Weaning without Pr-starter 不使用教保料断奶不使用教保料断奶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 2002280300320340360380400Length of Villis (m)绒毛长度绒毛长度04Da

23、ys after weaning 断奶后断奶后4天的变化天的变化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布拉迪布拉迪Fibers with high Hydratation capacity高水合能力的纤维Pigl

24、etsFeedsFeedsFatty 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：其次： 饲料的品质饲料的品质 Second : Feed Quality = More than nutritionnal levels Raw material quality 饲料原料的品质饲料原料的品质= Protein quality (high sensibility) 蛋白的品质蛋白的品质（较高的敏感性） Fiber quality (so

25、luble & insoluble) 纤维的品质纤维的品质（可溶&不可溶） Fat quality 脂肪的品质脂肪的品质 Starch quality 淀粉的品质淀粉的品质 Milk products quality 乳制品的品质乳制品的品质 Probiotics 益生菌的选择益生菌的选择 . Mycotoxins 霉菌毒素 Oxydations 氧化Slection of safety raw materials安全饲料原料的选择安全饲料原料的选择Limit soja meal 限制豆粕的使用 Technological Process, antinutritionnals factors

26、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425,0430,0435,0440,0445,0450,0455,0460,0465,0470,001234567FAT ingr (g/j)Hauteur villosits (m)Farine soya standardConc. protines soyaSlection of safety raw materials安全饲料原料的选择安全饲料原料的选择Soja meal 豆粕Soja meal t

27、reated 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 : 适宜的淀粉适宜的淀粉Trial Wheat Cooked wheatCereals variety and quality is important :Use of cooked wh

28、eat 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 TrialsAdapted starchs : 适宜的淀粉适宜的淀粉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Adaptat

29、ed Feeds :适宜的饲料适宜的饲料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种类型的淀粉针对不同的应用方案v Special gut protection, (days before weaning, 5 days after), 针对保护肠道，断奶前至断奶后针对保护肠道，断奶前至断奶后5天使用天使用v Spcial diahrea sensibility, low protein level v Spcial diahrea sensibility, normal protein

30、 level 针对腹泻敏感性选择淀粉，并配合不同蛋白水平针对腹泻敏感性选择淀粉，并配合不同蛋白水平v Spcial weaning 19 days, 针对不同断奶日龄选择淀粉针对不同断奶日龄选择淀粉v Spcial weaning 23 days, 并考虑是否需要着重保护菌群稳定性并考虑是否需要着重保护菌群稳定性v Spcial weaning 28 days , with flora protectionv Spcial weaning 28 days without high protectionv Spcial weaning 23 days, with antibiotic use (

31、probiotic inside)v 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 formulationSpcific feed free antibiotic trend :Raw materialsExemple of nutrient levels :Exemple of nutrient levels :Amino acids :Exemple of nu

32、trient levels :Vitamine AUI/kg17400Vitamine D3UI/kg116025 Hydchocalcifrolmg/kg0,021924equ. Vit D3876,96total D32036,96Vitamine Emg/kg174Polyphnols45Vitamine Cmg/kg150,8Vitamine B1mg/kg1,624Vitamine B2mg/kg5,8Pantothnate de Camg/kg17,4Niacinemg/kg29Vitamine B12mg/kg0,0406Vitamine B6mg/kg4,06Vitamine

33、K3mg/kg2,088Acide Foliquemg/kg1,16Biotinemg/kg0,116Chlorure de Cholinemg/kg290Btanemg/kg1044Fer mg/kg208,8Cuivre mg/kg162,4Zinc mg/kg0Manganse mg/kg63,8Iode (iodate)mg/kg1,16Cobalt (carbonate)mg/kg0Slnium (slnite)mg/kg0,2SlnoMthionine0,116Exemple of premix content :ExtractAdaptated solutions : exemp

34、le 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 : 适宜的益生菌适宜的益生菌Several probioti

35、cs 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

36、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371LEVUCELLSB :SPECIFIC PROBIOTIC YEAST布拉迪特殊的益生菌酵母特殊的益生菌酵母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) LE

37、VUCELL SB : Mode of Action 作用模式d) LEVUCELL SB : Main Applications and Role 主要应用与效果Third : 第三点第三点 381.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

38、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

39、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 鉴定与筛鉴定与筛选选391.a Bakers yeast : Maltose utilization Brewers yeast : Bitter Attenuation Wine yeast : Flavor, Color Distillers yeast: Fermenta

40、tion 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Identity and Selection 鉴定与筛选鉴定与筛选401.bOLD STORY BUT PROMISING FUTURE!Saccharomyces cerevisiae boulardii 酿酒酵母布拉氏亚种酿酒酵母布拉氏亚种1920. 发

41、现发现 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治疗人腹泻的药物上市 First form of boulardii active yeast recognized and marketed a

42、s 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 gu

43、t flora stabilizer in animal nutrition.1997. 完成分类学鉴定完成分类学鉴定boulardii was classified as a variety of Saccharomyces cerevisiae. Its genetic similarity and its functional differences are recognized.Today, 广泛使用广泛使用LEVUCELL SB is authorized and marketed in more than 44 countries.41 Research bibliography

44、booklet : Selection of 48 references related to S.c.boulardii in swine feeding. Abstracts in their original version.1.bS.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 applica

45、tions 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酿酒酵母布拉氏亚种这页可以去掉42第第1 1道防线道防线: : 肠道微生物肠道微生物强化优势菌群，如乳酸菌、纤维分解菌等第第2 2道防线道防线:

46、: 肠上皮细胞肠上皮细胞小肠通透性、粘膜厚度、抗菌肽分泌第第3 3道防线道防线: : 肠道免疫系统肠道免疫系统抗炎性、免疫细胞更新、白介素分泌LEVUCELLSB : Mode of ActionObjective : IMPROVEMENT OF GUT FUNCTION AND NATURAL SELF-DEFENSES改善肠道天然自体防御体系改善肠道天然自体防御体系1.cLEVUCELLSB : Mode of Action 作用模作用模式式431.c1st LINE OF DEFENSE :REGULATION OF INTESTINAL MICROFLORA第1道防线: 调整肠道微生

47、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清除残余氧清除残余氧Adhesion粘附作用粘附作用Neutralisation中和作用中和作用LEVUCELLSB : Mode of ActionUnique mode of action Neutralisation of microbial toxins (C.difficile, E.coli, .)特异性细菌毒素中和功能44 Faster O2 consumpti

48、on with LEVUCELLSB than with the other yeast strains布拉迪布拉迪 比其他酵母菌株的耗氧速度更快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 (SC

49、FA 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 consumption10 mL of a 0.1% resazurin solution+990 mL distilled water+5 g NaHCO3451.cIntestinal morphology肠道形态学肠道形态学Tight junctionsRegulation调节紧密连接调节紧密连接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第二道防线：维持肠道正常结构和功能

50、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 ActionLEVUCELLSB : Mode of Action46Improvement of physical barrier (gut)肠道物理屏障的改善肠道物理屏障的改善Control+LEVUCELLSBUt.Gand 绒毛长度 Villi (m)隐窝深度