CN112913978A - 一种仔猪肠道菌群调节剂 - Google Patents
一种仔猪肠道菌群调节剂 Download PDFInfo
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- CN112913978A CN112913978A CN202110256772.1A CN202110256772A CN112913978A CN 112913978 A CN112913978 A CN 112913978A CN 202110256772 A CN202110256772 A CN 202110256772A CN 112913978 A CN112913978 A CN 112913978A
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- vitamin
- feed additive
- senkyunolide
- galactomannan
- oligosaccharide
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Abstract
本发明涉及一种可调节仔猪肠道菌群的饲料添加剂，其中活性成分半乳甘露寡糖与洋川芎内酯A的重量比为(10:1)～(1:10)。与普通饲料添加剂相比，本发明的饲料添加剂制备方法简单，配伍合理，疗效显著，能够显著改善仔猪肠道菌群。由于添加剂中无西药成分，因此饲料添加剂无毒副作用，无耐药性。
Description
技术领域
本发明涉及饲料领域，具体涉及一种仔猪饲料添加剂。
背景技术
自20世纪中叶发现抗生素对动物的促生长作用以来，抗生素作为饲料添加剂在养殖业中得到了广泛应用；鉴于抗生素的负面效应以及人们对食品健康的日益关注，使得许多国家纷纷制定用以禁止抗生素和化学合成类药物使用的法律、法规。近年来，开始了安全、无毒、无副作用的抗生素替代品的开发研究。
研究证实，果寡糖可以提高猪、鸡、牛等动物的生产性能和疾病抵抗力；甘露寡糖可以提高鼠、猪、鸡、牛、水生动物等动物的生产性能和促进肠道中有益菌群(如双歧杆菌、乳酸杆菌等)增殖，而抑制有害菌(如大肠杆菌)的繁殖，并提高机体的免疫力。
半乳甘露寡糖(GMOS)又称半乳甘露低聚糖，它是由半乳甘露糖和葡萄糖通过糖苷键连接构成的寡糖，其为我国自主开发出的一个功能性寡糖品种。据报道，半乳甘露寡糖具有下列功能：一、通过优化肠道微生态系，形成以双歧杆菌和乳酸杆菌为优势菌的肠道菌群，对宿主起到有益的保健功能。二、对病原微生物有识别、粘附和排除作用。三、可为免疫调节剂，具有激活免疫系统应答的作用，这一特性也引起人们很大的兴趣。目前人们普遍认为甘露寡糖可以增强动物的非特异性免疫。近年来，多名研究人员验证了其在异育银鲫幼鱼、肉鸡、肉兔、早期断奶仔猪、常规断奶仔猪、生长育肥猪和泌乳母猪上的应用效果。
目前生产和制备GMOS的主要原料为半乳甘露聚糖胶，如田菁胶等。田菁胶来源于田菁种子的内胶乳，其化学组成是半乳甘露聚糖，相对分子质量约20万Da。含少量蛋白质，不溶于各种有机溶剂，易溶于水。田菁胶聚糖是由甘露糖单元构成主链，半乳糖单元形成支链，半乳糖与甘露糖单元之比为1：2。甘露糖单元通过α-1,4糖苷键连接，半乳糖单元通过β-1,6糖苷键连接在甘露糖主链上。半乳甘露寡糖的生产和制备主要是通过生物酶降解田菁胶得到。
用120尾异育银鲫饲养10周，结果表明，在基础饲料中添加0.2％GMOS能显著提高异育银鲫幼鱼增重率和特定生长率。降低饵料系数，同时能显著提高鱼体非特异性免疫功能。在增重率方面，0.1％组、0.2％组和0.3％组分别比空白对照组提高了2.33、30.0和18.1个百分点。在特定生长率方面，0.2％组和0.3％组分别比空白对照组提高了l9.8和9.5个百分点；而饵料系数0.1％组、0.2％组和0.3％组分别比空白对照组降低了l1.1、13.9和9.5个百分点。
试验结果表明，GMOS可一定程度替代肉鸡日粮抗生素(50mg/kg金霉素+40mg/kg盐霉素)，同时可显著提高血清新城疫抗体滴度水平，从而增强肉鸡体液和细胞免疫机能。
用30只50日龄新西兰肉兔饲养20d，结果表明，与添加0.005％喹乙醇的对照组相比，添加0.25％GMOS的试验组兔存活率提高了7.5％，料重比、腹泻率降低了6.1％和38.15％，此外屠宰后测得试验组的全净膛率比对照组提高了2.22％。
研究表明，GMOS对大肠杆菌引起的腹泻有预防作用，效果好于壳寡糖。GMOS能提高14日龄早期断奶仔猪生长潜能，表现在：能提高血清中类胰岛素生长因子-1(IGF-1)、生长激素浓度和肝脏、眼肌中IGF-1的基因表达水平，从而通过介导而起促生长作用：GMOS能提高14日龄早期断奶仔猪免疫功能。表现在：提高了仔猪血清中IgG、IgA、IgM、IL-2、IL-6及IL-1β免疫因子浓度，还提高了仔猪外周血单核细胞因子IL-1β、空肠黏膜IL-1β和肠系模淋巴结IL-lβ基因表达水平，从而增强机体细胞免疫和体液免疫。研究表明，饲喂不同浓度GMOS可降低早期断奶仔猪回肠微生物种群数量，抑制金黄色葡萄球菌、产气夹膜梭状芽孢杆菌等有害菌，促进部分乳酸杆菌、双歧杆菌等有益菌繁殖；在结肠段也可降低大肠杆菌数，明显提高结肠中乳酸杆菌和双歧杆菌数。
用72头28日龄断奶仔猪试验表明，日粮中添加0.12％GMOS可明显提高平均日采食量和平均日增重，并可降低腹泻发生率。试验表明，GMOS在9.4～15kg仔猪日粮中可完全取代饲用金霉素，并可获得更好生长性能，但在15～27kg仔猪上效果要逊色于75mg/kg金霉素。
用45头体重为25.3±2.3kg的杜×约二元猪饲喂94d，结果表明，在25～90kg生长育肥猪饲粮中添加一定比例酵母培养物和GMOS。可提高日增重、降低料重比、较少腹泻率。获得了良好的经济效益和社会效益。用0.10％GMOS饲喂生长猪96d，结果表明，与添加50mg/kg金霉素相比，可提高日增重，减少腹泻等消化道疾病发生率，但对气喘等呼吸道疾病没有改善效果；在血清生理生化指标方面，GMOS有降低血清中胆固醇和甘油三酯的趋势，可不同程度地降低血清谷草转氨酶、谷丙转氨酶活性，起到保肝、护肝作用，还可显著提高血清中GH、IL-2含量，进而起到促生长和提高免疫力的作用；在屠宰性能方面，GMOS有降低皮厚、背膘厚、眼肌面积和提高瘦肉率的趋势，对胴体直长、胴体斜长、屠宰率几乎没什么影响；在猪肉品质方面，GMOS可显著降低眼肌肉中吲哚含量，提高猪肉风味，此外还有提高系水力的趋势。
试验表明，当母猪饲粮营养水平相同且都不添加抗生素时，添加GMOS与未添加组相比，可显著提高母猪泌乳量、降低料乳比，对母猪内分泌机能也有调节作用；当添加抗生素时，GMOS同样可增加母猪泌乳量、降低料乳比，还可显著提高仔猪断奶个体重，改善效果明显好于对照组和吉他霉素，与维吉尼亚霉素相当；与对照组和2个抗生素组相比，半乳甘露寡糖组乳蛋白含量最高，平均提高幅度达14.9％。
中药制剂富含多种多糖等活性成分，在提高机体免疫力和抗病力方面有着显著的作用，现已证明数百种中草药有多方面的免疫活性，如促进免疫器官的生长发育，增强机体的细胞免疫和体液免疫，促进干扰素-γ、白细胞介素-2等细胞因子产生等作用，并且绿色、安全、不易产生耐药性。因此，中草药的开发利用已成为众望所归。
洋川芎内酯类属于内酯类化合物，主要包括洋川芎内酯A、洋川芎内酯H、洋川芎内酯I、洋川芎内酯D、洋川I芎内酯F、洋川芎内酯J、洋川芎内酯M、洋川芎内酯Q等，在中药川芎、当归等伞形科植物中含量较高，是主要活性成分及代谢成分，并具有多种药理活性。洋川芎内酯类化合物具有抗氧化损伤、抗炎镇痛、抗凝及抗血小板聚集、舒张血管、细胞毒等多种药理作用。
采用洋川芎内酯A对心肌细胞进行药物孵育55min后，再低氧2h，复氧1h，可使上清液中乳酸脱氢酶(LDH)明显减少、丙二醛(MDA)的含量降低、超氧化物歧化酶(SOD)的活性提高，与空白对照组相比还可降低心肌细胞内丝裂原活化蛋白激酶(MAPK)活性，表明洋川芎内酯A对于体外培养的乳鼠心肌细胞缺氧/复氧损伤具有保护作用，同时MAPK信号传导途径可能参与这一保护作用。
还有文献报道，洋川芎内酯A预处理对心脏微血管内皮细胞缺氧/复氧(H/R)具有保护作用，可使细胞存活数明显增加，细胞培养液中NO和一氧化氮合酶(iNOS)活性增加，内皮素活性降低，iNOS mRNA表达量增加而内皮素mRNA表达量则减少，其作用机制可能与上调内皮细胞中iNOS mRNA的表达和下调内皮素mRNA的表达有关。此外，采用Langedorff离体心脏灌流法，在恒温恒压条件下，用K-H灌流液与不同浓度的洋川芎内酯A灌流心脏。结果发现，洋川芎内酯A预处理可提高大鼠离体心脏停灌再灌期的冠脉流量，降低冠脉流出液中IL-1β的浓度及TXB2/6-Keto-PGF1α的比值，室性心动过速和心室颤动的发生率均降低，持续时间也缩短；同时心肌匀浆液中LDH、MDA的含量降低，SOD活性提高，表明洋川芎内酯A预处理对大鼠离体心脏缺血再灌注所致的血管内皮细胞损伤具有保护作用。
洋川芎内酯A对大鼠离体主动脉血管具有一定的舒张作用。其对PGF2α、苯肾上腺素、5-羟色胺、KC1诱导的血管收缩均具有抑制作用，并且腺嘌呤核苷酸环化酶抑制剂、可溶解的鸟甘酸环化酶抑制剂、非选择性的钾离子通道阻断剂四乙胺及去除内皮对洋川芎内酯A的舒张作用无明显影响，其具体作用机制有待进一步研究。
洋川芎内酯A对于结肠癌细胞HT-29具有细胞毒性，并能抑制其细胞增殖，IC50为(10.40±0.98)μg/mL。洋川芎内酯A对HT-29的存活性抑制较正常结肠细胞CCD-18Co的抑制性高，其IC50为(20.95±4.22)μg/mL，且洋川芎内酯A与丁苯酞、z-藁本内酯对HT-29细胞具有整合作用，并且对结肠癌细胞的抑制呈剂量依赖型。
目前，尚未见将半乳甘露寡糖与洋川芎内酯A组合作为调节肠道菌群的仔猪饲料添加剂的文献报道。
发明内容
本发明所要解决的技术问题是提供一种能有效调节肠道菌群，且毒副作用低的仔猪饲料添加剂。
本发明解决上述问题所采用的技术方案是，提供一种调节仔猪肠道菌群的饲料添加剂，其中活性成分半乳甘露寡糖与洋川芎内酯A的重量比为(10:1)～(1:10)。
优选的，所述饲料添加剂中半乳甘露寡糖与洋川芎内酯A的重量比为(9:1)～(1:1)。
更优选的，所述饲料添加剂中半乳甘露寡糖与洋川芎内酯A的重量比为(5:1)～(3:1)。
最优选的，所述饲料添加剂中半乳甘露寡糖与洋川芎内酯A的重量比为4:1。
优选的，所述饲料添加剂进一步包括其他活性成分。
更优选的，所述其他活性成分选自氨基酸、维生素等中的一种或多种。
最优选的，所述氨基酸为甘氨酸、丙氨酸、亮氨酸、缬氨酸、半胱氨酸、甲硫氨酸、苏氨酸、丝氨酸、苯丙氨酸、酪氨酸、色氨酸、蛋氨酸、谷氨酸、精氨酸和/或组氨酸。
最优选的，所述维生素为维生素A、维生素B1、维生素B2、维生素B3、维生素B5、维生素B6、维生素B12、维生素C、维生素D、维生素E、维生素H、维生素K和/或维生素P。
本发明还提供一种上述饲料添加剂在制备调节仔猪肠道菌群的饲料中的应用。
优选的，所述100重量份的饲料中，含有1-5重量％的饲料添加剂。
本发明具有积极有益的效果：
令人惊奇的是，本发明意外发现将半乳甘露寡糖与洋川芎内酯A组合使用，对于调节仔猪肠道菌群具有协同增效的效果，显著降低了活性成分的给药量，配合其他活性成分使用效果更佳。
与普通饲料添加剂相比，本发明的仔猪饲料添加剂制备方法简单，配伍合理，疗效显著，实验证明对调节仔猪肠道菌群效果良好；由于添加剂中无西药成分，因此饲料添加剂无毒副作用，无耐药性。
具体实施方式
下面结合实施例对本发明作更进一步的说明，但本发明的实施方式不限于此。下述实施例中所使用的实验方法如无特殊说明，均为常规方法。
实施例1、饲料添加剂的制备
取半乳甘露寡糖9重量份，洋川芎内酯A1重量份，混合后制成半乳甘露寡糖和洋川芎内酯A重量比为9:1的饲料添加剂S1。
取半乳甘露寡糖5重量份，洋川芎内酯A 5重量份，混合后制成半乳甘露寡糖和洋川芎内酯A重量比为1:1的饲料添加剂S2。
取半乳甘露寡糖8重量份，洋川芎内酯A 2重量份，混合后制成半乳甘露寡糖和洋川芎内酯A重量比为4:1的饲料添加剂S3。
实施例2、本发明饲料添加剂在调节断奶仔猪肠道菌群中的应用
1.试验动物及分组处理
选取杜洛克×(长白×大白)三元杂交28日龄断奶仔猪120头，所有公猪去势，按照体重、性别、血缘一致原则，随即分成6组，即本发明组合物组1-3(实施例1制备的饲料添加剂S1-S3)、半乳甘露寡糖组、洋川芎内酯A组和对照组，每组仔猪20头。在S1-S3组合物组、半乳甘露寡糖组、洋川芎内酯A组日粮中按照饲料总重量的1％添加上述饲料添加剂(每组加入的活性成分总重量相同)，对照组的日粮为不添加上述饲料添加剂的相同重量的猪饲料，各组仔猪每日投喂3次，试验期内自由采食，同时提供充足的饮水，其他各项日常管理措施按照养殖场常规管理进行，定期免疫和驱虫，试验期为30天。
2.肠道菌群测定
试验结束时，将各试验组仔猪进行屠宰，立即剖腹结扎取结肠段，用酒精棉球消毒各结扎口后，放入已灭菌塑料袋中，运到微生态实验室进行检测大肠杆菌、乳酸杆菌和双歧杆菌的数量。
培养基：大肠杆菌用伊红-美蓝培养基(EMB)，乳酸杆菌用MRS培养基，双歧杆菌用BLb培养基。
计数：采用平板涂布法进行细菌计数。以灭菌生理盐水进行倍比稀释为10-1、10-2、10-3---直到10-8，然后，用微量进样器从高倍稀释开始分别取稀释液0.02m1，滴入各选择培养基，每个样品做三个稀释度(大肠杆菌选10-4、10-5、10-6；乳酸杆菌选10-5、10-6、10-7；双歧杆菌选10-5、10-6、10-7)，每个稀释度做三个重复，取其平均数。接种后用曲玻棒推匀，大肠杆菌需氧培养24h，乳酸杆菌和双歧杆菌厌氧培养48h后，分别计数大肠杆菌、乳酸杆菌和双歧杆菌数。结果以cfu的常用对数值(logcfu)表示。
3.试验结果
各试验组饲料添加剂对断奶仔猪肠道菌群的影响见表1。
表1本发明饲料添加剂对断奶仔猪肠道菌群的影响(logcfu/g肠道内容物)
由实施例2可以看出，与对照组相比，本发明的饲料添加剂能显著降低仔猪肠道菌群中的大肠杆菌数量，提高乳酸杆菌和双歧杆菌的数量，是效果优异的仔猪肠道菌群调节剂。特别是半乳甘露寡糖和洋川芎内酯A的组合使用产生了协同效果，减少了药物的使用量，其中半乳甘露寡糖和洋川芎内酯A重量比为4:1的饲料添加剂效果最为突出，产生了难以预期的优异效果。
Claims (8)
1.一种可调节仔猪肠道菌群的饲料添加剂，其特征在于，其中活性成分半乳甘露寡糖与洋川芎内酯A的重量比为(10:1)～(1:10)。
2.根据权利要求1所述的饲料添加剂，其特征在于，半乳甘露寡糖与洋川芎内酯A的重量比为(9:1)～(1:1)。
3.根据权利要求2所述的饲料添加剂，其特征在于，半乳甘露寡糖与洋川芎内酯A的重量比为(5:1)～(3:1)。
4.根据权利要求1所述的饲料添加剂，其特征在于，所述饲料添加剂进一步包括其他活性成分。
5.根据权利要求4所述的饲料添加剂，其特征在于，所述其他活性成分选自氨基酸、维生素中的一种或两种。
6.根据权利要求5所述的饲料添加剂，其特征在于，所述氨基酸选自甘氨酸、丙氨酸、亮氨酸、缬氨酸、半胱氨酸、甲硫氨酸、苏氨酸、丝氨酸、苯丙氨酸、酪氨酸、色氨酸、蛋氨酸、谷氨酸、精氨酸和/或组氨酸，所述维生素选自维生素A、维生素B1、维生素B2、维生素B3、维生素B5、维生素B6、维生素B12、维生素C、维生素D、维生素E、维生素H、维生素K和/或维生素P。
7.权利要求1-6任一项所述的饲料添加剂在制备调节仔猪肠道菌群的饲料中的应用。
8.根据权利要求7所述的应用，其特征在于，所述100重量份的饲料中，含有1-5重量％的饲料添加剂。
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