CN107137528A - 防治海马烂尾病的药物及组合物 - Google Patents
防治海马烂尾病的药物及组合物 Download PDFInfo
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- CN107137528A CN107137528A CN201710392848.7A CN201710392848A CN107137528A CN 107137528 A CN107137528 A CN 107137528A CN 201710392848 A CN201710392848 A CN 201710392848A CN 107137528 A CN107137528 A CN 107137528A
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Abstract
本发明涉及水产养殖技术领域,尤其涉及防治海马烂尾病的药物及组合物。本发明提供的组合物由具有抑制海绵假交替单胞菌功能的活性物质组成,本发明通过体外实验证明,活性物质茶多酚、没食子酸、大蒜素、乳酸、聚赖氨酸皆能够具有抑制这种病原菌的作用,抑菌圈直径与大部分常用抗生素相当,且未表现出耐药性。本发明提供的组合物中,5种组分相互配合起到了良好的增效协同作用,在用量相当的条件下,组合物的抑菌圈直径显著大于单一组分的抑菌圈直径(p<0.05)。并且,本发明药物采用天然食药用成分,将通过海马代谢完全,无药物残留,无毒副作用,且不会产生抗药性,也不会对海马养殖水环境造成污染。
Description
技术领域
本发明涉及水产养殖技术领域,尤其涉及防治海马烂尾病的药物及组合物。
背景技术
海马为海龙科动物是刺鱼目海龙科暖海生数种小型鱼类的统称。海马作为一种名贵的中药材,在我国使用的历史悠久,早在《本草纲目》等多部医药著作中就有记载。海马因具有极高的药用和营养价值,享有“南方人参”之美誉,具有温肾壮阳、消肿散节和妇人催生等功效。
目前在热带和温带沿海的海草床、珊瑚礁、红树林和河口地区发现至少46个种的海马。2012年,世界自然保护联盟(IUCN)将38种海马列入濒危物种。作为缔约国,我国也已将海马列为国家二级保护动物。为了拯救海马这一濒危物种,并满足市场需求,除建立海马自然保护区,改善海马生存环境来保护海马自然资源外,通过人工养殖海马是海马保护的重要手段之一。我国自1957年以来已经开始养殖海马,然而目前海马养殖产业在国内推广并不是很好,限制其发展的主要瓶颈是养殖技术和病害。
海马养殖中,水质、水温、光线、比重及饵料等环境因子变化往往较大,海马由于不能适应这种环境的变化,病害发生较多,死亡率较高。据调查,海南省琼海市某海马种苗繁育基地,从2016年5月~8月,约30个养殖池几乎全部死亡,给该养殖基地造成巨大的损失。在同一时期,广东和福建沿海的养殖户也出现大面积死亡,死亡率高达80%~90%。而引起海马大规模死亡的主要原因是海马肠胃炎病和烂尾病。
海马烂尾病是由海绵假交替单胞菌(Pseudoalteromonas spongiae)导致的疾病,目前,针对海马的烂尾病的治疗方法还是以使用抗生素类药物为主,结合其他药物,如土霉素和聚维酮碘等,而长期使用抗生素类药物不但会使细菌产生抗药性,而且会影响海马健康和抗病害能力;药物残留不但会对环境产生危害,而且会产生严重的社会问题。近年来,从天然植物特别是药用植物中寻找纯天然抗生物药物已成为研究热点。
发明内容
有鉴于此,本发明要解决的技术问题在于提供防治海马烂尾病的药物及组合物。本发明提供的活性物质或组合物能够通过抑制海绵假交替单胞菌达到防治海马烂尾病的作用。
本发明提供了活性物质在制备抑制海绵假交替单胞菌的药物中的应用;
所述活性物质为茶多酚、没食子酸、大蒜素、乳酸、聚赖氨酸中的至少一种。
本发明实施例中,进行效果验证的菌株为:海绵假交替单胞菌Pseudoalteromonasspongiae HL11。
本发明还提供了活性物质在制备防治海马烂尾病的药物中的应用;所述活性物质为茶多酚、没食子酸、大蒜素、乳酸、聚赖氨酸中的至少一种。
本发明实施例中,进行效果验证的海马为大海马。
海马烂尾病由海绵假交替单胞菌引起,本发明通过体外实验证明,活性物质茶多酚、没食子酸、大蒜素、乳酸、聚赖氨酸皆能够具有抑制海绵假交替单胞菌的作用,抑菌圈直径与大部分常用抗生素相当,且未表现出耐药性。另有部分常见具有抑菌作用的活性物质(例如,Nisin、姜辣素等),未表现出对海绵假交替单胞菌的抑制作用。
本发明还提供了一种抑制海绵假交替单胞菌的组合物,该组合物由以下质量份的组分组成;
茶多酚0.5~7份;没食子酸1~3份;大蒜素1~3份;乳酸1~8份;聚赖氨酸2~12份。
本发明实施例中,组合物由如下质量份的组分组成:茶多酚0.5~5份;没食子酸1份;大蒜素1~3份;乳酸1~8份;聚赖氨酸2~10份。
经实验验证,本发明提供的组合物中,5种组分相互配合起到了良好的增效协同作用,在用量相当的条件下,组合物的抑菌圈直径显著大于单一组分的抑菌圈直径(p<0.05)。而在适宜配比下,增效协同作用更加显著,抑菌圈直径可达单一组分的2倍。
一些实施例中,本发明提供的组合物,由如下质量份的组分组成:茶多酚3份;没食子酸1份;大蒜素2份;乳酸1份;聚赖氨酸10份。
一些实施例中,本发明提供的组合物,由如下质量份的组分组成:茶多酚0.5份;没食子酸1份;大蒜素3份;乳酸8份;聚赖氨酸10份。
一些实施例中,本发明提供的组合物,由如下质量份的组分组成:茶多酚5份;没食子酸1份;大蒜素1份;乳酸5份;聚赖氨酸10份。
一些实施例中,本发明提供的组合物,由如下质量份的组分组成:茶多酚0.5份;没食子酸1份;大蒜素1份;乳酸1份;聚赖氨酸2份。
一些实施例中,本发明提供的组合物,由如下质量份的组分组成:茶多酚0.5份;没食子酸1份;大蒜素2份;乳酸5份;聚赖氨酸6份。
一些实施例中,本发明提供的组合物,由如下质量份的组分组成:茶多酚3份;没食子酸1份;大蒜素3份;乳酸5份;聚赖氨酸2份。
一些实施例中,本发明提供的组合物,由如下质量份的组分组成:茶多酚3份;没食子酸1份;大蒜素1份;乳酸8份;聚赖氨酸6份。
一些实施例中,本发明提供的组合物,由如下质量份的组分组成:茶多酚5份;没食子酸1份;大蒜素3份;乳酸1份;聚赖氨酸6份。
一些实施例中,本发明提供的组合物,由如下质量份的组分组成:茶多酚5份;没食子酸1份;大蒜素2份;乳酸8份;聚赖氨酸2份。
一些实施例中,本发明提供的组合物,由如下质量份的组分组成:茶多酚2.75份;没食子酸1份;大蒜素2份;乳酸1份;聚赖氨酸10份。
本发明提供的组合物能够用于防治由海绵假交替单胞菌引起的疾病,尤其适用于防治海马的烂尾病。
本发明提供组合物用于防治海马烂尾病的质量-体积浓度为45mg/L~270mg/L。
本发明还提供了防治海马烂尾病的药物,包括本发明所述的组合物和药学上可接受的辅料。
本发明提供的药物以药学上可接受的辅料制成各种剂型,以投放至饲养海马的水体中经过口服及接触防治海马的烂尾病。其剂型优选为:粉剂、片剂或胶囊剂。
本发明还提供了一种防治海马烂尾病的饲料,包括本发明所述的组合物和海马饵料。
本发明实施例中,饲料中所述组合物的质量-体积浓度为45mg/L~270mg/L。
所述体积为饲料总体积,本发明对海马饵料成分没有特殊要求,适宜海马各阶段自身的食性即可。
本发明还提供了一种海马的饲养方法,其特征在于,每年5~8月,以本发明所述的饲料饲喂海马,每天饲喂2次;其余时间以未添加药物的饵料饲喂海马。
本发明实验表明,健康大海马放入海马烂尾病原菌浓度为1×106cfu/mL的20L水体中,浸泡12h后,分别移入有效水体为70cm×50cm×50cm的玻璃缸中正常饲养和换水,连续观察30d。实验组采用本发明组合物,拌于饵料投喂,1天2次,对照组1采用等量土霉素,拌于饵料投喂;对照组2不采取措施,正常投喂,1天2次。对照组2第2d开始发病,海马起初是在水缸中出现平游和倒游,然后开始沉底,最后平躺于缸底,尾部向腹部弯曲;最终尾部的皮肤出现白色斑点、颜色变浅、发黄、变白死亡。至第10d,对照组2海马全部死亡。对照组1第8天才有发病症状,至第30d海马死亡率为60%。实验组海马在30d养殖期间,无发病现象。
本发明提供了防治海马烂尾病的药物及组合物,该组合物由具有抑制海绵假交替单胞菌功能的活性物质组成,本发明通过体外实验证明,活性物质茶多酚、没食子酸、大蒜素、乳酸、聚赖氨酸皆能够具有抑制海绵假交替单胞菌的作用,抑菌圈直径与大部分常用抗生素相当,且未表现出耐药性。另有部分常见具有抑菌作用的活性物质(例如,Nisin、姜辣素等),未表现出对海绵假交替单胞菌的抑制作用。经实验验证,本发明提供的组合物中,5种组分相互配合起到了良好的增效协同作用,在用量相当的条件下,组合物的抑菌圈直径显著大于单一组分的抑菌圈直径(p<0.05)。而在适宜配比下,增效协同作用更加显著,抑菌圈直径可达单一组分的2倍。而在同等抑菌效果下,组合物的绝对用量为单一组分的一半。并且,本发明药物采用天然食药用成分,将通过海马代谢完全,无药物残留,无毒副作用,且不会产生抗药性,也不会对海马养殖水环境造成污染。
具体实施方式
本发明提供防治海马烂尾病的药物及组合物,本领域技术人员可以借鉴本文内容,适当改进工艺参数实现。特别需要指出的是,所有类似的替换和改动对本领域技术人员来说是显而易见的,它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述,相关人员明显能在不脱离本发明内容、精神和范围内对本文的方法和应用进行改动或适当变更与组合,来实现和应用本发明技术。
茶多酚:又名茶单宁、茶鞣质,是茶叶中多酚类物质的总称,含黄烷醇类、花色苷类、黄酮类、黄酮醇类和酚酸类等成分,其中以属儿茶素类化合物的黄烷醇类含量最高,占茶多酚总量的60%-80%。茶多酚的抑菌谱广,对有细胞壁和无细胞壁的革兰氏阳性、阴性菌均有抑制作用,其对枯草杆菌、金黄色葡萄球菌、大肠杆菌、龋齿链球菌以及毛霉菌、青霉菌、赤霉菌、炭疽病菌、啤酒酵母菌有抑制作用。而且,茶多酚摄入人体后对人体有很好的生理效应;能清除人体内多余的自由基,能改进血管的渗透性能,降低血压,防止血糖升高,促进维生素C的吸收与同化,还有抗癌防龋、抗机体脂质氧化和抗辐射等作用。
没食子酸:又名五倍子酸、桔酸,为3,4,5-三羟基苯甲酸,为白色或淡黄色针状结晶或粉末,通常以一水合物的状态存在,是白色或微黄色针状晶体,是一种天然的多酚类化合物,它以游离酸形式或形成酯类化合物存在于五倍子、茶、没食子等植物中,是植物体中一类次生代谢产物,具有抗氧化、抑菌、防鍋齿等生物活性,可用于医药、有机合成和食品、农业、矿产等领域。没食子酸是多种植物的抑菌成分,对于枯草芽孢杆菌、大肠埃希菌、金黄色葡萄球菌、嗜水气单胞菌以及温和气单胞菌等都具有一定的抑菌作用。
大蒜素:大蒜为百合科葱属多年生草本植物,可作调味品、食品添加剂、饲料添加剂及美容化妆品的原料。大蒜具有杀菌、抑菌、抗病毒等医疗、保健功能。大蒜素是大蒜瓣和蒜精油中含硫化合物的总称,是大蒜的主要生物活性成分具有不同程度的抑菌作用,被誉为天然抑菌剂。蒜瓣被破碎、挤压时,由蒜氨酸与蒜苷酶接触,在磷酸吡哆醛催化下转化形成一类抑菌力最强的烯丙基二硫和三硫化物(亦称为大蒜新素)。
乳酸:广泛存在于食品特别是发酵食品中,如泡菜、腌菜、橄榄和某些肉、酸奶、奶酪及酸豆乳等发酵食品,多是采用乳酸菌种发酵所制。在乳酸菌发酵过程中,会产生许多有益于食品保藏的代谢产物,其中乳酸及少量醋酸是其中的主要代谢产物,它可以降低自身的pH值,从而有效抑制微生物的生长,达到抑菌和防腐效果。乳酸和乳酸盐被美国食品与药品管理局批准为“安全无毒物质(GRAS)”。乳酸和乳酸盐能抑制多种致病微生物的生长,如金黄色葡萄球菌、蜡样芽孢杆菌、埃希氏大肠杆菌和枯草芽孢杆菌等。
多聚赖氨酸:是一种由人体必须氨基酸L–赖氨酸的ε–氨基和另一L–赖氨酸的α–羧基形成ε–酰胺键连接而成的同聚物,是由一种白色链霉菌(Streptomyces albulus)发酵产生。多聚赖氨酸为阳离子表面活性物质,分子内部有疏水性的亚甲基,外部有亲水性的羧基和氨基。在水溶液中,游离的氨基带有正电荷,而正电荷有利于多聚赖氨酸与细菌细胞表面带负电的位点结合,因此具有良好的抑菌效果。目前,多聚赖氨酸已广泛应用到食品加工业的各个领域。
下面结合实施例,进一步阐述本发明:
实施例1
将目前,常用的抗生素或具有抗菌功能的物质进行海绵假交替单胞菌的抑菌圈实验。实验采用药物的体积为50μL。部分实验结果如表1~2:
表1常用抗生素抗菌效果
注:S,敏感;I,中介;R,耐药。
表2常用活性物质的抑菌圈直径(mm)
结果表明,聚赖氨酸、茶多酚、没食子酸、大蒜素和乳酸具有抑制海绵假交替单胞菌的作用,等浓度条件下,其效果与常用抗生素相当,甚至优于常用抗生素。而另一些常见具有抑菌功能的活性物质则未体现抑菌效果。
实施例2
按照表3配制组合物:
表3:组合物配方
序号 | 茶多酚(mg) | 乳酸(mg) | 大蒜素(mg) | 多聚赖氨酸(mg) | 没食子酸(mg) |
1 | 5 | 10 | 10 | 20 | 10 |
2 | 5 | 50 | 20 | 60 | 10 |
3 | 5 | 80 | 30 | 100 | 10 |
4 | 30 | 10 | 20 | 100 | 10 |
5 | 30 | 50 | 30 | 20 | 10 |
6 | 30 | 80 | 10 | 60 | 10 |
7 | 50 | 10 | 30 | 60 | 10 |
8 | 50 | 50 | 10 | 100 | 10 |
9 | 50 | 80 | 20 | 20 | 10 |
10 | 27.5 | 10 | 20 | 100 | 10 |
将表3组合物分别溶解至1L,各取50μL进行对海绵假交替单胞菌的抑菌圈实验,每组实验重复3次,测量结果取平均值结果如表4:
表4各组合物对海绵假交替单胞菌抑菌圈直径
结果显示,组合物效果显著优于单一组分的效果(p<0.05),例如,第2~4、6~8、10号组方的组合物,其效果皆显著优于单一组分(p<0.05),各组分的复配起到了增效协同作用。而不当的配比,仅能维持相似的抑菌效果,甚至降低对病原菌的抑制作用。效果最佳的组方为4号组方和第3号组方,其抑菌效果显著优于其他组方。
实施例3
健康大海马放入海马烂尾病原菌浓度为1×106cfu/mL的20L水体中,浸泡12h后,分别移入有效水体为70cm×50cm×50cm的玻璃缸中正常饲养和换水,连续观察30d。
实验组采用本发明组合物(分别给予2~4、6~8、10号组方),拌于饵料投喂,1天2次,
对照组1采用等量土霉素,拌于饵料投喂,1天2次;
对照组2不采取措施,正常投喂,1天2次。
结果显示,对照组2第2d开始发病,海马起初是在水缸中出现平游和倒游,然后开始沉底,最后平躺于缸底,尾部向腹部弯曲;最终尾部的皮肤出现白色斑点、颜色变浅、发黄、变白死亡。至第10d,对照组2海马全部死亡。对照组1第8天才有发病症状,至第30d海马死亡率为60%。实验组海马在30d养殖期间,无发病现象。
以上仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
Claims (10)
1.活性物质在制备抑制海绵假交替单胞菌的药物中的应用;
所述活性物质为茶多酚、没食子酸、大蒜素、乳酸、聚赖氨酸中的至少一种。
2.活性物质在制备防治海马烂尾病的药物中的应用;
所述活性物质为茶多酚、没食子酸、大蒜素、乳酸、聚赖氨酸中的至少一种。
3.一种抑制海绵假交替单胞菌的组合物,其特征在于,由以下质量份的组分组成;
茶多酚0.5~7份;没食子酸1~3份;大蒜素1~3份;乳酸1~8份;聚赖氨酸2~12份。
4.根据权利要求3所述的组合物,其特征在于,由如下质量份的组分组成:茶多酚3份;没食子酸1份;大蒜素2份;乳酸1份;聚赖氨酸10份。
5.根据权利要求3所述的组合物,其特征在于,由如下质量份的组分组成:茶多酚0.5份;没食子酸1份;大蒜素3份;乳酸8份;聚赖氨酸10份。
6.根据权利要求3所述的组合物,其特征在于,由如下质量份的组分组成:茶多酚5份;没食子酸1份;大蒜素1份;乳酸5份;聚赖氨酸10份。
7.一种防治海马烂尾病的药物,其特征在于,包括权利要求3~6任一项所述的组合物和药学上可接受的辅料。
8.一种防治海马烂尾病的饲料,其特征在于,包括权利要求3~6任一项所述的组合物和海马饵料。
9.根据权利要求8所述的饲料,其特征在于,其中所述组合物的质量-体积浓度为45mg/L~270mg/L。
10.一种海马的饲养方法,其特征在于,每年5~8月,以权利要求8或9所述的饲料饲喂海马,每天饲喂2次;其余时间以未添加药物的饵料饲喂海马。
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CN116711659A (zh) * | 2023-06-13 | 2023-09-08 | 中国科学院南海海洋研究所 | 假交替单胞菌在促进珊瑚幼虫高产生物膜和诱导珊瑚幼虫附着生长中的应用 |
CN117448230A (zh) * | 2023-11-03 | 2024-01-26 | 广东海洋大学 | 一株海绵假交替单胞菌及其在对虾养殖中的应用 |
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CN116711659B (zh) * | 2023-06-13 | 2024-01-05 | 中国科学院南海海洋研究所 | 假交替单胞菌在促进珊瑚幼虫高产生物膜和诱导珊瑚幼虫附着生长中的应用 |
CN117448230A (zh) * | 2023-11-03 | 2024-01-26 | 广东海洋大学 | 一株海绵假交替单胞菌及其在对虾养殖中的应用 |
CN117448230B (zh) * | 2023-11-03 | 2024-03-26 | 广东海洋大学 | 一株海绵假交替单胞菌及其在对虾养殖中的应用 |
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