CN106616029B - 一种提高虹鳟抗病能力的饲料添加剂及其制备方法 - Google Patents
一种提高虹鳟抗病能力的饲料添加剂及其制备方法 Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Marine Sciences & Fisheries (AREA)
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Fodder In General (AREA)
- Feed For Specific Animals (AREA)
Abstract
本发明涉及一种提高虹鳟抗病能力的饲料添加剂,按重量份数计包括:β‑1,3葡聚糖20‑35份、低分子壳寡糖15‑25份、维生素C 10‑22份、柠檬酸5‑10份、蛋氨酸锌3‑6份、5‑甲基‑2‑(4‑甲氧基苯基)‑4‑噁唑甲酸乙酯0.5‑2份。添加本发明所提供的饲料添加剂后可以有效降低了虹鳟感染寄生虫病、病毒性及细菌性疾病的几率,增强其防病抗病能力,同时可大幅提升养殖虹鳟的成活率及品质。
Description
技术领域
本发明属于水产养殖技术中的水产饲料添加剂领域,具体涉及到一种提高虹鳟抗病能力的饲料添加剂。
背景技术
虹鳟,俗称鳟鱼,原产地北美西部,在中国为引进品种,分布在北京、黑龙江、山东、山西、辽宁、吉林、陕西等地。因其肉里无刺,易于消化,富含维生素D、铁元素的含量、高度不饱和脂肪酸特别是22炭6烯酸和20炭5烯酸,前者具有助脑发育成长,活化脑细胞,延缓脑老化的功效,后者具有降低血脂,防止动脉硬化,心肌梗塞和脑中风的功效,鱼皮富含蛋白质,具有美容和强健筋骨的功效,除此还含有能供给人体必需的氨基酸、矿物质、维生素A和维生素D等人体所需元素,因此虹鳟常被称为水中人参,具有较高的营养价值。
随着社会消费水平的提高,虹鳟的消费市场也在逐步扩大,虹鳟的养殖市场具有广阔的发展前景。在虹鳟过养殖过程中,目前主要的病害来自于病毒性、细菌性疾病,如鳃病、肠道疾病等,除传染性疾病外,商业化养殖的主要病害威胁还来自于寄生虫的攻击,即寄生虫病,如鱼虱病等。
目前的防治主要是使用抗生素为主,但使用大量抗生素不仅污染环境,而且还严重人体健康。为了提高虹鳟的抗病性和减少抗生素的使用,可在虹鳟饲料中添加其他药用成分以提高抗病性,从而提升虹鳟的成活率和产量。
发明内容
本发明的目的在于克服现有技术的不足之处,提供一种提高虹鳟抗病能力的饲料添加剂,可增强该鱼种的免疫力并促进其生长。
本发明实现目的的技术方案如下:
一种苯基噁唑类化合物5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯在防治鱼类寄生虫病饲料添加剂中的应用;尤其是在防治虹鳟鱼鱼虱病害饲料添加剂中的应用。
一种提高虹鳟抗病能力的饲料添加剂,按重量份数计包括:β-1,3葡聚糖20-35份、低分子壳寡糖15-25份、维生素C 10-15份、柠檬酸5-10份、蛋氨酸锌3-6份、5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯0.5-9份。
进一步地上述提高虹鳟抗病能力的饲料添加剂,按重量份数计包括:β-1,3葡聚糖30份、低分子壳寡糖20份、维生素C 12份、柠檬酸8份、蛋氨酸锌5份、5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯7份。
此外,本发明还提供了上述提高虹鳟抗病能力的饲料添加剂的制备方法,具体步骤包括:将上述原料各自按量称取,先将β-1,3葡聚糖、低分子壳寡糖、维生素C、蛋氨酸锌、5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯分别研磨成粉、过100目筛后混合,再加入柠檬酸,混合均匀后制得所述虹鳟抗病能力的饲料添加剂。
虹鳟养殖目前采用的是集约化养殖,养殖过程中又极容易发生细菌性疾病及寄生虫病害,当发生寄生虫病时,寄生虫通过咬食鳞片、上皮和粘膜来损害鱼,当感染严重时,这些寄生虫还会损害到下层真皮,因此即使除去了寄生虫,但是仍多发继发感染和涝渍,极大的影响了虹鳟的产量。本发明提供的相应饲料添加剂应用于虹鳟人工养殖过程中,能有效的抑制病毒、细菌、寄生虫病害,增强鱼体的免疫力,大大提升了该品类鱼的抗病能力,进而提升虹鳟的成活率和产量。
本发明配方中添加了少量的5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯,可有效抑制虹鳟的寄生虫病。5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯是一种已知的苯基噁唑类化合物,该化合物分子式为C14H15NO4,分子量为261.2,浅黄色固体,目前该化合物的应用研究多见于抗癌药物领域的研制,而未见在水产养殖尤其是在鳟鱼类疾病病害防治中应用的相关研究和报道。本发明人通过大量的实验发现在一定浓度添加范围内,5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯能迅速抑制寄生虫如鱼虱的活性,在24小时的测定中基本能完全杀死相应鱼虱,并对鱼体无害。
同时,本发明还添加了β-1,3葡聚糖、低分子壳寡糖、维生素C等,β-1,3葡聚糖可以有效提升鱼体的免疫能力;添加低分子壳寡糖后虹鳟的吞噬活力等有所增强,同时增强了对杀鲑气单胞菌的抵抗力;维生素的添加不仅能提高饲料的营养成分,还对鱼类出血病、烂鳃病有一定的预防作用。
本发明提供的上述原料互相匹配后,经过研发人员对虹鳟各生长指标进行反复试验论证后发现,增加该饲料添加剂后有效降低了虹鳟感染寄生虫病、病毒性及细菌性疾病的几率,增强了其防病抗病能力,大幅提升了养殖虹鳟的成活率及品质;同时因减少了大量抗生素的使用,极大程度的降低了抗生素带来的鱼体残留、环境污染等危害。
具体实施方式
下面通过具体实施例对本发明作进一步详述,以下实施例只是描述性的,不是限定性的,不能以此限定本发明的保护范围。
实施例1
称取β-1,3葡聚糖20g、低分子壳寡糖15g、维生素C 15g、蛋氨酸锌4g份、5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯0.5g,分别研磨成粉,过100目筛,混合,再添加入柠檬酸6kg,混合均匀,即得本发明所提供的提高虹鳟抗病能力的饲料添加剂。
其中5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯可参照已有的合成方式制得:将适量乙酰乙酸乙酯溶于NaNO2的醋酸溶液中,搅拌冷却到5-7℃条件下,进行亚硝化反应,反应完成后加入乙醚,过滤收集析出的结晶,用甲醇-乙醚按体积比1:5再重结晶,得到α-羟亚氨基化合物;然后在氯化氢气体饱和的醋酸溶液中,α-羟亚氨基化合物与对甲氧基苯甲醛缩合,生成不稳定的中间体氮氧化合物;运用催化加氢还原中间体氮氧化合物,即可获得化合物:5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯。
实施例2
将β-1,3葡聚糖30g、低分子壳寡糖20g、维生素C20g、蛋氨酸锌5g、5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯0.8g分别研磨成粉,过100目筛,混合,再添加入柠檬酸8g,混合均匀,即得本发明所提供的提高虹鳟抗病能力的饲料添加剂。其中5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯的合成方式参照实施例1中所示。
实验对比
1、5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯毒性实验
5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯已见有在人体及动物体内进行抗癌实验的研究,研究表明其对细胞无毒性作用;而该化合物应用于水产养殖,尤其是应用于防治鱼虱病这种寄生虫病的研究还鲜见报道。下列实验是针对其对鱼虱尤其是海虱防治的体内实验及鱼体的耐受性实验。
取出30条在养殖场中被感染鱼虱的虹鳟,分成三组每组各10条,一组为空白组,其他两组为实验组,为每组虹鳟感染的鱼虱计数,将三组鱼转入充气良好的海水箱中,保持48小时以便适应环境,在加入5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯之前禁食24小时,两组对照组虹鳟中一组以添加5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯1.0ppm测试浓度进行处置,另一组以2ppm的浓度进行处置,饲喂后保持24小时,然后对死亡和存活的寄生虫进行计数。24小时后实验表明,空白组虹鳟感染鱼虱数量未见明显减少,而两组对比组感染鱼虱数量均接近0;同时采用5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯进行长效实验,对染病虹鳟体表鱼虱病达到了几乎100%的治愈,时效可保持到4个月左右的时间内,且无任何鱼死亡现象,被虹鳟很好的耐受,而实验中作为对照的空白组(即染病虹鳟,未加以治疗),长达4个月中出现了20%的死亡率,同时鱼体较大面积受损率高达70%。上述实验充分说明5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯能有效的控制虹鳟养殖中常见的鱼虱寄生虫病,并能被鱼体很好的耐受。
2、采用本发明饲料添加剂后虹鳟的免疫指标测定
选用80条虹鳟为八组(每组10条)进行对比实验,其中四组为健康虹鳟、四组为明显染有寄生虫病害的虹鳟;在健康虹鳟组和染有病害虹鳟组中,各有一组为空白对照组,即仅添加基础饲料,各试验组:试验1为在基础饲料中添加实施例1所示饲料添加剂,试验组2是在基础饲料中添加实施例2所示饲料添加剂,两组饲料添加剂添加到基础饲料中的比例以质量百分比计均为0.3%;试验组3为在基础饲料中添加了实施例2中除5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯以外的饲料添加剂组分(即仅添加了将β-1,3葡聚糖30g、低分子壳寡糖20g、维生素C20g、蛋氨酸锌5g、柠檬酸8g),且该饲料添加剂添加到基础饲料中的比例以质量百分比计也为0.3%。投喂时间持续60天后,测定虹鳟鱼血细胞的吞噬活性、溶菌酶活力。免疫指标测定:
1)采血
用MS-222(150mg/L)麻醉鱼体,1%肝素湿润的注射器尾部血管采血。将每尾鱼的血液分为2份,其中1份血液首先在室温下静置1h待其凝固,置冰箱4℃保存4h,4℃,4000r/min离心10min,收集血清用于溶菌酶测定;另1份用肝素抗凝,制备抗凝血,供测定白细胞吞噬活性。
2)吞噬细胞吞噬功能的测定
在抗凝血中加入0.25ml金黄色葡萄球菌悬液,充分混匀后于28℃下水浴1h,水浴期间每隔10min摇匀1次。再加入0.15mol/L冷NaCl溶液0.8ml,2000r/min离心4min,弃去上清液。然后用吸管吸取血细胞涂片,每个血样涂5片,甲醇固定5~7min,蒸馏水冲洗,Giemsa染色1~5h,水洗晾干后镜检。吞噬活性分别以吞噬百分比(PP)和吞噬指数(PI)表示:PP%=(100个吞噬细胞中参与吞噬的细胞数/100)×100%;PI%=(吞噬细胞内的细菌总数/参与吞噬的吞噬细胞数)×100%。3)溶菌酶(LSZ)活力的测定
以溶壁微球菌为底物,用0.1mol/ml磷酸缓冲液(pH6.4)将溶壁微球菌配成0.3mg/ml的菌悬液,每个样品取3ml菌悬液,吸取0.04ml血清加入菌悬液中,混匀后立即用UV-2000分光光度计于570nm下测定AO值。然后在37℃水浴中温浴30min后,取出置于4℃水浴中10min以终止反应,测定其于570nm处的光密度值A,按下式计算溶菌酶活力:U=(AO-A)/AO。
测定结果详见下表:结果表明添加了实施例1、实施例2所示饲料添加剂的饲料均能显著地提高虹鳟鱼的吞噬活力、血清溶菌酶活力也显著地提高,与空白对照组相比,尤其是吞噬活力指标值差异均极显著;且相较而言试验组2即添加实施例2饲料添加剂组的各项指标也明显好于试验组1。
健康组和染病组虹鳟相比,除了两组空白对照相外,两大组中的试验1组相对比、试验2组相对比,差异性不大,也侧面验证了本发明能较有效的防治虹鳟寄生虫病害,提升其免疫力及抗病能力;大组内试验2组的各项指标性能明显好于其他组别。
以上对本发明的较佳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 (5)
1.一种提高虹鳟抗病能力的饲料添加剂,其特征在于按重量份数计包括:β-1,3葡聚糖20-35份、低分子壳寡糖15-25份、维生素C10-22份、柠檬酸5-10份、蛋氨酸锌3-6份、5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯0.5-2份。
2.根据权利要求1所述的提高虹鳟抗病能力的饲料添加剂,其特征在于按重量份数计包括:β-1,3葡聚糖30份、低分子壳寡糖20份、维生素C12份、柠檬酸8份、蛋氨酸锌5份、5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯0.8份。
3.根据权利要求1或2所述提高虹鳟抗病能力的饲料添加剂的制备方法,其特征在于包括如下步骤:将上述β-1,3葡聚糖、低分子壳寡糖、维生素C、蛋氨酸锌、5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯分别研磨成粉、过100目筛后混合,再加入柠檬酸,混合均匀后制得所述虹鳟抗病能力的饲料添加剂。
4.苯基噁唑类化合物5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯在防治鱼类寄生虫病饲料添加剂中的应用。
5.根据权利要求4所述的苯基噁唑类化合物5-甲基-2-(4-甲氧基苯基)-4-噁唑甲酸乙酯在防治鱼类寄生虫病饲料添加剂中的应用,其特征在于在防治虹鳟鱼鱼虱病害饲料添加剂中的应用。
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