CN115105490A - 一种高效抑制产气荚膜梭菌的植物精油组合物及应用 - Google Patents
一种高效抑制产气荚膜梭菌的植物精油组合物及应用 Download PDFInfo
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Abstract
本发明专利公开了一种高效抑制产气荚膜梭菌的植物精油组合物及应用,具体涉及畜牧养殖药物的技术领域。包括如下重量组份的原料:40‑60份的肉桂醛、4‑8份的α蒎烯、10‑20份的月桂酰甘氨酸和1‑4份的乙二胺四乙酸。采用本发明技术方案解决了禁用抗生素生长促进剂后有效抑制产气荚膜梭菌的问题,可用于畜禽养殖,提高畜牧养殖业的效益。
Description
技术领域
本发明涉及畜牧养殖药物的技术领域,特别涉及一种高效抑制产气荚膜梭菌的植物精油组合物及应用。
背景技术
抗生素生长促进剂(Antibiotic Growth Promoter,AGP)在国内外的应用已有60多年的历史,为提高畜牧养殖的效益提供了不可或缺的保障。但低剂量AGP的持续性添加所引发的细菌耐药性大发展,以及畜禽产品的药物残留问题也愈来愈引起人们的关注。逐步限制乃至最终全面禁止AGP的使用,已经在全球范围内成为行业和社会共识。
我国是抗生素产销第一大国,其中超过一半被用于畜牧养殖,作为AGP使用的抗生素每年超过10万吨,占全球使用量的30%以上。尽管近年来以家禽、生猪为代表的畜禽养殖业集约化程度和养殖水平均有了大幅度的提升,但在种苗的有效净化工作尚未全面完成之前,全面禁用AGP任重而道远。
尽管国内外对AGP替代技术的研究已有很多,所涉及的替代品包括酸化剂、植物提取物、微生态制剂、酶制剂、短链脂肪酸、抗菌肽等等,但迄今为止,尚无任何一种能够达到现在所使用的AGP的效能。
目前AGP替代品的筛选试验,绝大多数以大肠杆菌、沙门氏菌等G-为目标菌。实际上,肠道内产气荚膜梭菌(Clostridium perfringens)的感染才是AGP发挥作用的真正目标菌。正是其所产生的α、β外毒素上调了肠黏膜的炎症反应水平,一方面影响了肠黏膜对营养物质的吸收,另一方面加大了动物机体用于免疫防御的资源消耗。然而,遗憾的是,目前以产气荚膜梭菌作为目标菌的报道很少。
发明内容
本发明意在提供一种高效抑制产气荚膜梭菌的植物精油组合物及应用,解决了禁用抗生素生长促进剂后有效抑制产气荚膜梭菌的问题。
为了达到上述目的,本发明的技术方案如下:一种高效抑制产气荚膜梭菌的植物精油组合物,包括如下重量组份的原料:40-60份的肉桂醛、4-8份的α蒎烯、10-20份的月桂酰甘氨酸和1-4份的乙二胺四乙酸。
技术方案的原理及效果:在α蒎烯的存在下,月桂酰甘氨酸促进了肉桂醛对产气荚膜梭菌细胞壁、细胞膜的渗透性能,大幅度提高了肉桂醛的抑菌活性;乙二胺四乙酸通过络合细菌表面的金属离子,增强了月桂酰甘氨酸抑制产气荚膜梭菌产生和分泌外毒素的性能。
进一步的,包括如下重量组份的原料:50-60份的肉桂醛、6-8份的α蒎烯、10-15份的月桂酰甘氨酸和1-2份的乙二胺四乙酸。
进一步的,所述植物精油组合物包括如下步骤:
S1、预备权利要求1或2的重量组份的原料肉桂醛、α蒎烯、月桂酰甘氨酸和乙二胺四乙酸;
S2、将肉桂醛加热至45℃-55℃,然后加入三分之二组份的月桂酰甘氨酸搅拌至溶解,再加入乙二胺四乙酸继续搅拌至溶解,制得第一溶液;
S3、将剩余三分之一组份的月桂酰甘氨酸加入α蒎烯搅拌溶解,制得第二溶液;
S4、将第二溶液缓慢加入第一溶液,然后搅拌均匀得到植物精油组合物。
进一步的,将植物精油组合物通过后喷涂工艺添加至颗粒饲料、或通过轻质二氧化硅吸附后用于饲料混合加工,所述植物精油组合物剂量:15-20ppm,以肉桂醛计。
本发明提供的另一种技术方案:将上述的一种高效抑制产气荚膜梭菌的植物精油组合物应用于畜牧养殖业。
与现有技术相比,本方案的有益效果:
本方案通过肉桂醛与合适种类的乳化剂以特定比例配合,结合特定比例的其它活性成分,实现了将其MIC值降低到10μg/ml以下。并且可以有效减少产气荚膜梭菌外毒素的产生和分泌。
附图说明
图1是本实施例中不同浓度肉桂醛作用于产气荚膜梭菌后的流式细胞术检测结果图。
具体实施方式
下面通过具体实施方式对本发明作进一步详细的说明:
实施例
一种高效抑制产气荚膜梭菌的植物精油组合物,包括如下重量组份的原料:40-60份的肉桂醛、4-8份的α蒎烯、10-20份的月桂酰甘氨酸和1-4份的乙二胺四乙酸。
本实施例中原料采用50-60份的肉桂醛、6-8份的α蒎烯、10-15份的月桂酰甘氨酸和1-2份的乙二胺四乙酸。
植物精油组合物包括如下步骤:
S1、预备上述的重量组份的原料:50-60份的肉桂醛、6-8份的α蒎烯、10-15份的月桂酰甘氨酸和1-2份的乙二胺四乙酸。
S2、将50-60份的肉桂醛加热至45℃-55℃,然后加入月桂酰甘氨酸总量中三分之二组份的月桂酰甘氨酸,在80-120rpm的条件下搅拌至溶解,再加入乙二胺四乙酸继续搅拌至溶解,制得第一溶液。
S3、将剩余三分之一组份的月桂酰甘氨酸加入α蒎烯在80-120rpm的条件下搅拌溶解,搅拌过程中保持常温状态,制得第二溶液。
S4、将第二溶液缓慢加入第一溶液,然后搅拌均匀得到植物精油组合物。
将步骤S4中制备的植物精油组合物通过后喷涂工艺添加至颗粒饲料、或通过轻质二氧化硅吸附后用于饲料混合加工,所述植物精油组合物剂量:15-20ppm,以肉桂醛计。
将上述植物精油组合物与饲料混合加工后应用于畜牧养殖业,主要用于生猪、肉鸡、肉鸭、蛋鸡等动物养殖。
产气荚膜梭菌的抑制效果:
将本方案中单个原料、不同原料之间的混合物和本方案配制成梯度浓度的溶液,测定其对产气荚膜梭菌的最低抑菌浓度(MIC)。土霉素作为传统抗生素促生长剂对照,得到如表1所示的结果。
表1:各原料对产气荚膜梭菌的MIC测定结果(μg/mL)
注:/代表MIC>100mg/mL,通常认为不具备抑菌活性。
通过表1可知,本方案的组合物对产气荚膜梭菌的抑制效果最优,相较于传统的抗生素促生长剂的土霉素而言,其效果明显增强。
不同浓度肉桂醛作用于产气荚膜梭菌后的流式细胞术检测:
肉桂醛、抑菌活性较高的组合物以及本发明所采用的方案均以MIC的浓度作用于产气荚膜梭菌6h后,通过流式细胞仪检测细菌凋亡比例。结果如图1所示,其中图1A为未经处理的产气荚膜梭菌;图1B为肉桂醛处理的产气荚膜梭菌;图1C为肉桂醛+月桂酰肌氨酸钠+α蒎烯处理的产气荚膜梭菌;图1D为本方案处理的产气荚膜梭菌。
由图1可见,正常培养的产气荚膜梭菌基本不存在凋亡、细胞膜受损等情况(图1A所示),肉桂醛诱导了产气荚膜梭菌的凋亡,MIC的肉桂醛作用后诱导了59.5%的细菌坏死,9.87%的细菌细胞膜完整性受到破坏,9.03%的细菌细胞膜轻微受损(图1B);肉桂醛+月桂酰肌氨酸钠+α蒎烯作用后诱导了58.6%的细菌坏死,21.3%细菌细胞膜完整性受到破坏,15%的细菌细胞膜轻微受损(图1C);本发明采用的方案中肉桂醛组合物作用后诱导了63%的细菌坏死,14.8%细菌细胞膜完整性受到破坏,15%的细菌细胞膜轻微受损(图1D)。
通过上述情况可得如下结论:尽管肉桂醛及其一般组合物,诱导产气荚膜梭菌凋亡的比例比较接近,但本发明采用的方案中肉桂醛的使用剂量不到肉桂醛单独使用或简单组合剂量的十分之一,而用量越小成本越低,同时也避免肉桂醛的刺激性引起的适口性差的弊端。。
以上的仅是本发明的实施例,方案中公知的具体结构和/或特性等常识在此未作过多描述。应当指出,对于本领域的技术人员来说,在不脱离本发明结构的前提下,还可以作出若干变形和改进,这些也应该视为本发明的保护范围,这些都不会影响本发明实施的效果和专利的实用性。本申请要求的保护范围应当以其权利要求的内容为准,说明书中的具体实施方式等记载可以用于解释权利要求的内容。
Claims (5)
1.一种高效抑制产气荚膜梭菌的植物精油组合物,其特征在于:包括如下重量组份的原料:40-60份的肉桂醛、4-8份的α蒎烯、10-20份的月桂酰甘氨酸和1-4份的乙二胺四乙酸。
2.根据权利要求1所述的一种高效抑制产气荚膜梭菌的植物精油组合物,其特征在于:包括如下重量组份的原料:50-60份的肉桂醛、6-8份的α蒎烯、10-15份的月桂酰甘氨酸和1-2份的乙二胺四乙酸。
3.根据权利要求1或2中任一项所述的一种高效抑制产气荚膜梭菌的植物精油组合物,其特征在于:所述植物精油组合物包括如下步骤:
S1、预备权利要求1或2的重量组份的原料肉桂醛、α蒎烯、月桂酰甘氨酸和乙二胺四乙酸;
S2、将肉桂醛加热至45℃-55℃,然后加入三分之二组份的月桂酰甘氨酸搅拌至溶解,再加入乙二胺四乙酸继续搅拌至溶解,制得第一溶液;
S3、将剩余三分之一组份的月桂酰甘氨酸加入α蒎烯搅拌溶解,制得第二溶液;
S4、将第二溶液缓慢加入第一溶液,然后搅拌均匀得到植物精油组合物。
4.根据权利要求3所述的一种高效抑制产气荚膜梭菌的植物精油组合物,其特征在于:将植物精油组合物通过后喷涂工艺添加至颗粒饲料、或通过轻质二氧化硅吸附后用于饲料混合加工,所述植物精油组合物剂量:15-20ppm,以肉桂醛计。
5.将权利要求3的一种高效抑制产气荚膜梭菌的植物精油组合物应用于畜牧养殖业。
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