CN111034855A - 一种增强动物呼吸道免疫的组合物及方法和其应用 - Google Patents
一种增强动物呼吸道免疫的组合物及方法和其应用 Download PDFInfo
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- CN111034855A CN111034855A CN201911407185.7A CN201911407185A CN111034855A CN 111034855 A CN111034855 A CN 111034855A CN 201911407185 A CN201911407185 A CN 201911407185A CN 111034855 A CN111034855 A CN 111034855A
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Abstract
本发明提供一种增强动物呼吸道免疫的组合物,由添加进饲料的拌料添加剂和喷涂禽舍的喷雾复合菌制剂组成。拌料添加剂的原料包括:复合发酵中药70‑95份,维生素A 0.2‑0.5份,维生素C 4‑8份,维生素E 1‑2份。其中复合发酵中药包括:甘草:桑叶:薄荷:苦杏仁:金银花:黄芪=2:1:1:1:1:1。喷雾复合菌制剂原料为:枯草芽孢杆菌:地衣芽孢杆菌:植物乳杆菌:粪肠球菌=1:1:1:1,喷雾浓度为2mL/m3。本发明根据呼吸道粘膜的特殊性,采用拌料和喷雾两种方式,共同发挥作用,增强呼吸道粘膜的免疫功能,全面防治动物呼吸道疾病。
Description
技术领域
本发明涉及饲料添加剂领域,更具体地说涉及一种增强动物呼吸道免疫的组合物及方法和其应用。
背景技术
近年来,随着规模场的快速增加,畜禽呼吸道疾病越来越引起人们的重视,其预防和治疗也变得尤为重要。比如,支原体的感染,是动物养殖中较为棘手的疾病。支原体的感染后,一方面呼吸道上皮细胞和绒毛细胞破坏,降低呼吸道粘膜屏障作用,另一方面,呼吸道的免疫***受到损伤,增加机体对各种病原菌、病毒的敏感性,引发继发性感染。
动物机体除了全身性的免疫机能以外,还包括上呼吸道和消化道粘膜的局部免疫。这两者为机体的天然门户,保护呼吸道和肠道免疫,可以阻碍病原菌的侵入。目前,改善呼吸道粘膜,增强呼吸道的免疫的措施主要包括以下几种:A.加强饲养管理,改善饲养环境:畜舍环境内的有害气体、饲料粉尘颗粒、病原微生物等会对动物的呼吸道粘膜造成很大的威胁,保证良好的饲养密度,饲养密度维持在30-33kg/m2,适度通风,定期打扫圈舍卫生,消毒,全进全出对畜禽呼吸道有保护作用。B.合理使用疫苗和用药:免疫和用药可以辅助行的增强机体的呼吸道粘膜免疫。合理适度选择抗菌药,增强畜禽呼吸道免疫;C.使用饲料添加剂:中药、益生菌、维生素等饲料添加剂可以促进畜禽粘膜的发育,促进损伤粘膜的修复,提高呼吸道的免疫功能。
本发明通过两种方式共同来改善呼吸道粘膜免疫,添加剂A添加剂A包括:复合发酵中药、维生素A、维生素C、维生素E,作为拌料添加剂,长期使用,从动物机体着手,增强呼吸道粘膜免疫力。添加剂B的原料包括:枯草芽孢杆菌、地衣芽孢杆菌、植物乳杆菌、粪肠球菌,为喷雾制剂,从***环境着手,能够降低环境中的总需氧菌、大肠杆菌、金黄色葡萄球菌数量,改善鸡舍环境。
中国专利公报CN108740340A公布了一种预防动物呼吸道疾病的动物饲料,通过在饲料中添加中药添加剂,该中药添加剂中包括大青叶、甘草、川大黄、玄参和连翘,实现对猪呼吸道疾病的防治,中药原药单一粉碎后添加,其有效成分释放有限,防治呼吸道疾病缓慢。
中国专利公报CN107952062A公布了一种动物用呼吸道防治药物及其制备方法,该动物用呼吸道防治药物包括匹多莫德、辣木、褪黑素、微藻和复合益生菌,本发明结合抗生素和益生菌的共同作用,治疗呼吸道疾病。
发明内容
本发明克服了现有技术中的不足,提供了一种增强动物呼吸道免疫的组合物及方法和其应用。
本发明的目的通过下述技术方案予以实现。
一种增强动物呼吸道免疫的组合物,由添加进饲料的拌料添加剂和喷涂禽舍的喷雾复合菌制剂组成。
进一步,所述拌料添加剂各组分的重量份数为:复合发酵中药70-95份,维生素A0.2-0.5份,维生素C4-8份,维生素E1-2份。
进一步,所述复合发酵中药中各中药原料添加比例为甘草:桑叶:薄荷:苦杏仁:金银花:黄芪=2:1:1:1:1:1。
进一步,所述喷雾复合菌制剂中各原料组分的添加比例为枯草芽孢杆菌:地衣芽孢杆菌:植物乳杆菌:粪肠球菌=1:1:1:1。
增强动物呼吸道免疫的组合物的制备方法,包括以下步骤:
S1:制备复合发酵中药:按照复合中药组方取中药混合,按照所述复合中药质量的0.5%接种复合菌种,然后发酵;发酵温度为25℃左右,时间为一周,每天翻动2-3次,发酵后减压真空干燥。发酵结束后,有发酵清香,水分达到39%,PH≤4.0,总活菌数≥2*108CFU/g。
S2:制备拌料添加剂:将所述S1中得到的所述复合发酵中药与维生素混合;
S3:制备喷雾复合菌制剂:按比例称取枯草芽孢杆菌、地衣芽孢杆菌、植物乳杆菌、粪肠球菌溶于水中,充分搅拌,成悬浮状。
进一步,所述S1中所述复合中药组方的各中药原料添加比例为甘草:桑叶:薄荷:苦杏仁:金银花:黄芪=2:1:1:1:1:1。
进一步,所述S1中发酵时优选的复合菌种为枯草芽孢杆菌、酵母菌、丁酸梭菌和粪肠球菌混合物,总活菌数≥2*1010。
进一步,所述S1中制备复合中药的步骤为:
第一步,将所述复合中药组方的各中药原料粉碎,过筛30目;
第二步,所述第一步中的混合物料在温度121℃的条件下加温45分钟,水蒸气灭菌,灭菌后的水分控制在40%-60%;
所述S3中所述喷雾复合菌制剂中各原料组分的添加比例为枯草芽孢杆菌:地衣芽孢杆菌:植物乳杆菌:粪肠球菌=1:1:1:1。
增强动物呼吸道免疫的饲料添加剂在饲料添加剂及肉禽舍抑菌方面的应用。
本发明的有益效果为:
固体发酵复合中药具有资源丰富、绿色无污染、生物学功能突出、替抗减抗的优势,目前在动物养殖中使用较为广泛。本发明主要结合动物呼吸道免疫的特殊性,精选针对呼吸道粘膜有改善的几味中药进行复配。甘草:补脾益气、清热解毒、祛痰止咳,对呼吸道疾病有较明显的防治作用;桑叶:抗菌作用鲜桑叶煎剂体外实验,对金黄色葡萄球菌、白喉杆菌和炭疸杆菌均有较强的抗菌作用,对大肠杆菌、伤寒杆菌、痢疾杆菌、绿脓杆菌也有一定的抗菌作用。薄荷:利咽、疏散风热、清利头目,疏肝行气。苦杏仁:降气止咳平喘,多用于咳嗽气喘、胸满痰多等。金银花:清热解毒、抗菌消炎、增强免疫功能。黄芪:作为免疫促进剂和调节剂,具有抗病毒、抗应激、抗氧化的作用。
复合维生素对呼吸道粘膜免疫具有重要作用。维生素A促进皮肤和粘膜增生,稳定上皮细胞细胞膜,参与呼吸道黏膜免疫调控,能够缓解脂多糖和氨气对呼吸道的应激作用,提高呼吸道免疫功能。维生素C提高机体的杀菌和抗病毒能力,构成胶原蛋白、维护皮肤、呼吸道粘膜的完整性和坚固性;维生素E抗氧化剂,可以增强肝脏的解毒能力,提高机体免疫力。
添加剂B中的原料组成为复合益生菌制剂,枯草芽孢杆菌和地衣芽孢杆菌是芽孢乳酸菌,耐受性强,喷雾在空气中,可抑制环境中微生物的生长,降低舍内空气所携带病原微生物。植物乳杆菌和粪肠球菌属于乳酸菌,产酸能力较强,可以抑制大肠杆菌的定植,调节动物呼吸道菌群的多样性,提高呼吸道的免疫力。复合益生菌进入动物体内,还可在动物代谢过程中,分泌蛋白酶、纤维素酶等,促进营养物质消化吸收,减少有害气体的排除,改善畜禽养殖环境。
与现有技术相比,本发明具有的有益效果为:①复合饲料添加剂,分为拌料制剂和喷雾制剂,从内到外,全方位提高动物呼吸道免疫;②中药组方采取益生菌发酵方式处理,催化中药有效成分的释放,增强其生物活性;③添加剂B采用新颖的喷雾制剂的方式,充分结合呼吸道粘膜易损机制,有效增强呼吸道粘膜免疫;④复合维生素的添加,使多种维生素协同作用,更好的保护呼吸道粘膜;⑤使用绿色添加剂,避免出现抗生素耐药性等问题,改善动物和人类健康;⑥改善动物福利,增加动物养殖收入。
具体实施方式
下面通过具体的实施例对本发明的技术方案作进一步的说明。
实施例1
一种增强动物呼吸道免疫的饲料添加剂,包括两个部分,第一部分为拌料添加剂,为添加剂A;第二部分为喷雾复合菌制剂,为添加剂B。添加剂A的原料包括:复合发酵中药95份,维生素A0.25份,维生素C4份,维生素E1份。其中复合发酵中药包括:甘草:桑叶:薄荷:苦杏仁:金银花:黄芪=2:1:1:1:1:1。添加剂B为复合菌制剂,原料为:枯草芽孢杆菌(1500亿CFU/g):地衣芽孢杆菌(1500亿CFU/g):植物乳杆菌(1500亿CFU/g):粪肠球菌(1500亿CFU/g)=1:1:1:1,喷雾浓度为2mL/m3。
验证本发明一种增强动物呼吸道免疫的饲料添加剂对于肉禽舍环境的改善效果。申请人在山东某肉鸡场进行试验,随机选择4栋通风、光照、饲养面积一致的圈舍进行试验,2栋进行喷雾试验,按比例称取枯草芽孢杆菌、地衣芽孢杆菌、植物乳杆菌、粪肠球菌共计20g,溶于20kg的水中,充分搅拌,成悬浮状,即可。每天早晚各喷雾一次。试验周期为42天,试验时间为2019年5月20日—2019年7月1日。
饲养管试验理按照养殖场正常规范进行操作鸡,按照常规程序进行免疫,7日龄进行新城疫和传支二联苗点眼滴鼻免疫,21日龄进行新城疫饮水免疫。每周采用空气采集器采集圈舍气体,选点位置为前、中、后个3个点,共9个点,每个点采集3次平行,每次采集固定相同位置。总需氧菌、大肠杆菌和金黄色葡萄球菌分别以营养琼脂培养基、伊红美兰培养基和甘露醇高盐培养基为采样介质。采样器采样流量设置为28.3L/min,总需氧菌和金黄色葡萄球菌采集时间为2-3min,大肠杆菌采集时间为2-4min。采样期间关闭风机,样品采集完成后,立即放入恒温培养箱培养,总需氧菌和大肠杆菌培养18~24h,金黄色葡萄球菌培养24~36h,之后进行鉴定计数。
试验数据采用SPSS12.0统计软件对试验数据进行统计分析,经单因子方差比较,结果以“平均值±标准差”表示。试验结果如下表1、表2:
表1添加剂对鸡舍空气微生物含量的影响(lgCFU/m3)
备注:同列肩注不同字母,表示差异显著(p<0.05)。
试验结果表明,随着饲养日龄的增长,圈舍的空气质量降低,有害微生物数量增加,21日龄,试验组的总需氧菌、大肠杆菌、金黄色葡萄球菌数量显著低于对照组(p<0.05)。42日龄,试验组的总需氧菌、大肠杆菌、金黄色葡萄球菌数量显著低于对照组(p<0.05)
表2添加剂对不同圈舍肉鸡***物微生物含量的影响(lgCFU/m3)
备注:同列肩注不同字母,表示差异显著(p<0.05)。
由上表可知,随着饲养日龄的增长,肉鸡***物的微生物数量增加,21日龄,试验组的总需氧菌、大肠杆菌、金黄色葡萄球菌数量显著低于对照组(p<0.05)。42日龄,试验组的总需氧菌、大肠杆菌、金黄色葡萄球菌数量显著低于对照组(p<0.05)。
以上实施例表明,本发明添加剂可以明显降低圈舍空气和***物中总需氧菌、大肠杆菌和金黄色葡萄球菌等微生物含量,净化动物养殖环境,降低呼吸道疾病发生率。
实施例2
一种增强动物呼吸道免疫的饲料添加剂,包括两个部分,第一部分为拌料添加剂,为添加剂A;第二部分为喷雾复合菌制剂,为添加剂B。添加剂A的原料包括:复合发酵中药92份,维生素A0.35份,维生素C6份,维生素E2份。其中复合发酵中药包括:甘草:桑叶:薄荷:苦杏仁:金银花:黄芪=2:1:1:1:1:1。添加剂B为复合菌制剂,原料为:枯草芽孢杆菌(1500亿CFU/g):地衣芽孢杆菌(1500亿CFU/g):植物乳杆菌(1500亿CFU/g):粪肠球菌(1500亿CFU/g)=1:1:1:1,喷雾浓度为2mL/m3。
验证本发明一种增强动物呼吸道免疫的饲料添加剂对肉鸡呼吸道黏膜免疫的影响。试验在陕西省某肉鸡养殖场进行,肉鸡品种为AA肉鸡,随机选择两栋进行试验,一组为对照组,一组为试验组,试验组添加本发明添加剂。每栋肉鸡存栏为3000羽。
试验方法:21日龄时,每个处理随机选取10只采样。翅静脉采血,4℃下3000r/min,离心10min,分离血浆后.20℃冷冻保存待测,静脉采血后处死;取长度大约lcm的气管10%***固定、脱水透明、浸蜡、包埋、切片;气管切开后,留置针置入,以lml PBS灌洗,反复3次,轻轻按摩动物胸部20-30s后,回收肺泡灌洗液黏蛋白质,回收率在80%-90%,3000r/min,离心5rain,上清保存在20℃冰箱中待测定。取气管和肺分装,置于-80℃速冻保存待测。
试验结果如下表3、表4、表5:
表3添加剂对肉鸡血清IgA的影响
组别 | IgA(ng/mL) |
对照组 | 820.76±25.18<sup>b</sup> |
试验组 | 916.79±23.96<sup>a</sup> |
备注:同列肩注不同字母,表示差异显著(p<0.05)。
试验结果表明,本发明添加剂可以提高肉鸡的血清IgA含量,增强肉鸡呼吸道免疫。
表4添加剂对肺泡灌洗液黏蛋白质浓度的影响
组别 | 肺泡灌洗液黏蛋白质(ug/mL) |
对照组 | 612.35±35.73<sup>b</sup> |
试验组 | 873.77±25.18<sup>a</sup> |
备注:同列肩注不同字母,表示差异显著(p<0.05)。
试验结果表明,本发明添加剂可以增加肺泡灌洗液黏蛋白质的浓度,促进呼吸道粘膜的生长。
表5对肺泡灌洗液IgA的含量的影响
组别 | 肺泡灌洗液IgA(ng/mL) |
对照组 | 601.46±36.12<sup>b</sup> |
试验组 | 769.46±25.54<sup>a</sup> |
备注:同列肩注不同字母,表示差异显著(p<0.05)。
试验结果表明,本发明添加剂可以增加肺泡灌洗液IgA的含量,增强呼吸道免疫力,保护呼吸道免受应激和疾病影响。
以上对本发明的两个实施例进行了详细说明,但所述内容仅为本发明的较佳实施例,不能被认为用于限定本发明的实施范围。凡依本发明申请范围所作的均等变化与改进等,均应仍归属于本发明的专利涵盖范围之内。
Claims (10)
1.一种增强动物呼吸道免疫的组合物,其特征在于:由添加进饲料的拌料添加剂和喷涂禽舍的喷雾复合菌制剂组成。
2.根据权利要求1所述的增强动物呼吸道免疫的组合物,其特征在于:所述拌料添加剂各组分的重量份数为:复合发酵中药70-95份,维生素A 0.2-0.5份,维生素C 4-8份,维生素E1-2份。
3.根据权利要求2所述的增强动物呼吸道免疫的组合物,其特征在于:所述复合发酵中药中各中药原料添加比例为甘草:桑叶:薄荷:苦杏仁:金银花:黄芪=2:1:1:1:1:1。
4.根据权利要求1所述的增强动物呼吸道免疫的组合物,其特征在于:所述喷雾复合菌制剂中各原料组分的添加比例为枯草芽孢杆菌:地衣芽孢杆菌:植物乳杆菌:粪肠球菌=1:1:1:1。
5.如权利要求1-4任一所述的增强动物呼吸道免疫的组合物的制备方法,其特征在于:包括以下步骤:
S1:制备复合发酵中药:按照复合中药组方取中药混合,按照所述复合中药质量的0.5%接种复合菌种,然后发酵,发酵至有发酵清香,水分达到39%,PH≤4.0,总活菌数≥2*108CFU/g;
S2:制备拌料添加剂:将所述S1中得到的所述复合发酵中药与维生素混合;
S3:制备喷雾复合菌制剂:按比例称取枯草芽孢杆菌、地衣芽孢杆菌、植物乳杆菌、粪肠球菌溶于水中,充分搅拌,成悬浮状。
6.根据权利要求5所述的制备方法,其特征在于:所述S1中所述复合中药组方的各中药原料添加比例为甘草:桑叶:薄荷:苦杏仁:金银花:黄芪=2:1:1:1:1:1。
7.根据权利要求5所述的制备方法,其特征在于:所述S1中发酵时采用的复合菌种为枯草芽孢杆菌、酵母菌、丁酸梭菌和粪肠球菌混合物,总活菌数≥2*1010。
8.根据权利要求5所述的制备方法,其特征在于:所述S1中制备复合中药的步骤为:
第一步,将所述复合中药组方的各中药原料粉碎,过筛30目;
第二步,所述第一步中的混合物料在温度121℃的条件下加温45分钟,水蒸气灭菌,灭菌后的水分控制在40%-60%。
9.根据权利要求5所述的制备方法,其特征在于:所述S3中所述喷雾复合菌制剂中各原料组分的添加比例为枯草芽孢杆菌:地衣芽孢杆菌:植物乳杆菌:粪肠球菌=1:1:1:1。
10.利用如权利要求1-4任一所述的增强动物呼吸道免疫的饲料添加剂在饲料添加剂及肉禽舍抑菌方面的应用。
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