CN107347752A - 一种红螯螯虾孵卵繁育方法 - Google Patents
一种红螯螯虾孵卵繁育方法 Download PDFInfo
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- CN107347752A CN107347752A CN201710457422.5A CN201710457422A CN107347752A CN 107347752 A CN107347752 A CN 107347752A CN 201710457422 A CN201710457422 A CN 201710457422A CN 107347752 A CN107347752 A CN 107347752A
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Classifications
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- 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
- A01K61/10—Culture of aquatic animals of fish
- A01K61/17—Hatching, e.g. incubators
-
- 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
- A01K61/50—Culture of aquatic animals of shellfish
- A01K61/59—Culture of aquatic animals of shellfish of crustaceans, e.g. lobsters or shrimps
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- A—HUMAN NECESSITIES
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Abstract
本发明公开了一种红螯螯虾孵卵繁育方法,包括亲虾培育,脱卵,消毒,着床,孵化。其中所用消毒剂成分及其重量份为:球孢链霉菌发酵液3‑8份,蟹壳提取物4‑7份,柠檬烯5‑10份,β‑水芹烯1‑3份,石竹烯2‑5份,柠檬酸3‑6份。有益效果为:本发明公开的方法所用的消毒剂对病原体毒性强,少量的消毒剂便可起到很好的效果,虾卵对该消毒剂的耐受性好,在杀死病原体的同时,不影响孵化率,所繁育的仔虾成活率高,品质好,繁育步骤简单,操作方便,实用性强,具有良好的经济效益。
Description
技术领域
本发明涉及水产养殖技术领域,尤其是涉及一种红螯螯虾孵卵繁育方法。
背景技术
红螯螯虾属甲壳纲、十足目、长尾亚目、拟河虾科、光壳虾属,原产于澳大利亚北部的热带地区和新几内亚南部的溪流中,因其外形酷似海水龙虾,又只生活在淡水中,故俗称澳洲淡水龙虾。红螯螯虾具有个体大、生长快、食性杂、易饲养、病害少等养殖性能优势;同时,因其肉味鲜美、富含低胆固醇蛋白质,作为营养保健食品而风靡欧美等国,其市场前景广阔。目前,美国、法国、西班牙、南非、东南亚等许多国家引进了此虾,并进行工厂化育苗及养殖。
现有技术如申请公布号为CN 103314908 A的中国发明专利,公开了一种红螯螯虾卵离体培育技术,该技术中虾卵经过消毒,分级孵化等过程得所需的虾苗,该发明提高虾苗的成活率,但提高的程度有限,难以满足市场对虾苗的要求。
发明内容
本发明的目的在于提供一种红螯螯虾孵卵繁育方法,方法步骤简单,操作方便,实用性强,极大地提高了虾苗的成活率和品质,虾卵对所选用的消毒剂的耐受性好,在杀死病原体的同时,不影响孵化率。
本发明针对背景技术中提到的问题,采取的技术方案为:一种红螯螯虾孵卵繁育方法,首先是亲虾培育:亲虾按雌雄比例3-4∶1进行专池培育;接着是脱卵:采用消毒后的镊子将虾卵从亲虾腹部分离;紧接着是消毒:取1-3份消毒剂,加入40-45份水,搅拌混合得消毒液,取2-5份虾卵,加入制备的消毒液,浸泡3-6h,取出;最后是孵化:将消毒后的卵虾置于孵化器中,孵化密度为125-130粒/mL,孵化池内溶解氧保持在10-13mg/L,孵化温度保持在25-27℃,孵化液由水、水解蛋白、α-亚麻酸、复合维生素、葡萄糖、葡萄糖酸钙、硫酸锌、海藻酸钠按100/4/0.3/0.4/7/4/0.2/7的重量比混合制成。其中,消毒剂的成分及其重量份为:球孢链霉菌发酵液3-8份,蟹壳提取物4-7份,柠檬烯5-10份,β-水芹烯1-3份,石竹烯2-5份,柠檬酸3-6份。其中,球孢链霉菌购买自上海一研生物科技有限公司。虾卵对所制备的消毒剂耐受性强,且其功效不是各组分作用的简单叠加,而是显著的协同增效,提高了消毒剂对病原体的药效,降低了用药量,符合绿色环保的要求。
作为优选,球孢链霉菌发酵液的制备方法为:取1-3份球孢链霉菌,加入9-12份无菌生理盐水得菌体悬浮液,向悬浮液中加入4-6份活性短肽,再接种到40-50份的察氏培养液中,置于20-23℃、170-175r/min的恒温摇床上培养4-5d,离心分离,取上清液得链霉菌发酵液。其中,察氏培养液的成分及其重量份为:5-7份蔗糖,1-3份FeSO4,1-2份NaNO3,2-3份K2HPO4,1-4份MgSO4·7H2O,5-7份KCl,10-13份蒸馏水。该培养基含有丰富的营养物质,为球孢链霉菌的新陈代谢提供了有利条件,所得的球孢链霉菌发酵液所含的抗生素含量高,抑菌活性强,提高了所制备的消毒剂的功效。
作为优选,球孢链霉菌发酵液的制备过程中所加入的活性短肽的氨基酸序列为:HSHACHPGKLCVCVNCSR。发酵液要达到一定的抑菌活性,需要长时间的摇床培养,加入的活性短肽改变了细胞膜的通透性,提高了营养物的传质速度,促进了胞内的新陈代谢,缩短了培养时间。
作为优选,蟹壳提取物的制备方法为:取10-15份蟹壳,用10-13份质量分数为12%-15%HCl溶液浸泡5-7h,过滤,水洗至中性,置于8-10份温度为质量分数为35%-40%NaOH溶液中,在温度为96-100℃下搅拌15-17h,离心分离,将所得固体水洗至中性,得蟹壳提取物。该方法制备步骤少,其蟹壳提取物中所含的甲壳素和壳聚糖含量高,增强了消毒剂对病原体的作用。
作为优选,消毒剂的制备方法为:取3-8份球孢链霉菌发酵液,加入3-6份柠檬酸,再加入4-7份蟹壳提取物,搅拌20-25min,再加入5-10份柠檬烯,1-3份β-水芹烯,2-5份石竹烯,在35-40℃下搅拌1.5-2h,得消毒剂。该条件下,各种成分可以充分混合,使消毒剂更加均匀和稳定,提高了产品在实际应用过程中的效果。
与现有技术相比,本发明的优点在于:本发明公开的一种红螯螯虾孵卵繁育方法,过程中所用的消毒剂对病原体毒性强,少量的消毒剂便可起到很好的效果,虾卵对该消毒剂的耐受性好,在杀死病原体的同时,不影响孵化率,所繁育的仔虾成活率高,品质好,繁育步骤简单,操作方便,实用性强,具有良好的经济效益。
具体实施方式
下面通过实施例对本发明做进一步说明:
实施例1:
一种红螯螯虾孵卵繁育方法,包括:
1)亲虾培育:亲虾按雌雄比例3∶1进行专池培育;
2)脱卵:采用消毒后的镊子将虾卵从亲虾腹部分离;
3)消毒:取1份消毒剂,加入40份水,搅拌混合的消毒液,取2份虾卵,加入制备的消毒液,浸泡3h,取出;
4)孵化:将消毒后的卵虾置于孵化器中,孵化密度为125粒/mL,孵化池内溶解氧保持在10mg/L,孵化温度保持在25℃,孵化液由水、水解蛋白、α-亚麻酸、复合维生素、葡萄糖、葡萄糖酸钙、硫酸锌、海藻酸钠按100/4/0.3/0.4/7/4/0.2/7的重量比混合制成。
步骤3)中消毒剂的成分及其重量份为:球孢链霉菌发酵液3份,蟹壳提取物4份,柠檬烯5份,β-水芹烯1份,石竹烯2份,柠檬酸3份。该消毒剂的制备方法包括:首先是球孢链霉菌发酵液的制备:点燃酒精灯,将接种环通过火焰灼烧灭菌,将灭菌的接种环伸入菌种管内,先将环接触管内壁,达到冷却的目的,然后再挑取1份菌苔,将接种环退出菌种管,迅速伸入9份无菌生理盐水得菌体悬浮液,向悬浮液中加入4份活性短肽,再接种到40份的察氏培养液中,置于20℃、170r/min的恒温摇床上培养4d,在6800r/min下离心10min,取上清液得链霉菌发酵液。其中,察氏培养液的成分及其重量份为:5份蔗糖,1份FeSO4,1份NaNO3,2份K2HPO4,1份MgSO4·7H2O,5份KCl,10份蒸馏水。该培养基含有丰富的营养物质,为球孢链霉菌的新陈代谢提供了有利条件,所得的球孢链霉菌发酵液所含的抗生素含量高,抑菌活性强,提高了所制备的消毒剂的功效;接着是蟹壳提取物的制备:向容器中加入10份蟹壳,再加入10份质量分数为12%HCl溶液浸泡5h,过滤,水洗至中性,置于8份温度为质量分数为35%NaOH溶液中,用水浴锅控制温度为96℃下,搅拌反应15h,在4200r/min下离心8min,将所得固体水洗至中性,得蟹壳提取物。该方法制备步骤少,其蟹壳提取物中所含的甲壳素和壳聚糖含量高,增强了消毒剂对病原体的作用;最后复配制备消毒剂:取3份球孢链霉菌发酵液,加入3份柠檬酸,再加入4份蟹壳提取物,搅拌20min,再加入5份柠檬烯,1份β-水芹烯,2份石竹烯,在35℃下搅拌1.5h,得消毒剂。该条件下,各种成分可以充分混合,使消毒剂更加均匀和稳定,提高了产品在实际应用过程中的效果。
实施例2:
一种红螯螯虾孵卵繁育方法,包括:
1)亲虾培育:亲虾按雌雄比例5∶1进行专池培育;
2)脱卵:采用消毒后的镊子将虾卵从亲虾腹部分离;
3)消毒:取3份消毒剂,加入45份水,搅拌混合的消毒液,取5份虾卵,加入制备的消毒液,浸泡6h,取出;
4)孵化:将消毒后的卵虾置于孵化器中,孵化密度为130粒/mL,孵化池内溶解氧保持在13mg/L,孵化温度保持在27℃,孵化液由水、水解蛋白、α-亚麻酸、复合维生素、葡萄糖、葡萄糖酸钙、硫酸锌、海藻酸钠按100/4/0.3/0.4/7/4/0.2/7的重量比混合制成。
步骤3)中消毒剂的成分及其重量份为:球孢链霉菌发酵液8份,蟹壳提取物7份,柠檬烯10份,β-水芹烯3份,石竹烯5份,柠檬酸6份。该消毒剂的制备方法包括:
1)球孢链霉菌发酵液的制备:取3份球孢链霉菌,加入12份无菌生理盐水得菌体悬浮液,向悬浮液中加入6份活性短肽,再接种到50份的察氏培养液中,置于23℃、175r/min的恒温摇床上培养5d,离心分离,取上清液得链霉菌发酵液。其中,察氏培养液的成分及其重量份为:7份蔗糖,3份FeSO4,2份NaNO3, 3份K2HPO4,4份MgSO4·7H2O,7份KCl,13份蒸馏水。该培养基含有丰富的营养物质,为球孢链霉菌的新陈代谢提供了有利条件,所得的球孢链霉菌发酵液所含的抗生素含量高,抑菌活性强,提高了所制备的消毒剂的功效;
2)蟹壳提取物的制备:取15份蟹壳,用13份质量分数为15%HCl溶液浸泡7h,过滤,水洗至中性,置于10份温度为质量分数为40%NaOH溶液中,在温度为100℃下搅拌17h,离心分离,将所得固体水洗至中性,干燥得蟹壳提取物。该方法制备步骤少,其蟹壳提取物中所含的甲壳素和壳聚糖含量高,增强了消毒剂对病原体的作用;
3)消毒剂的制备:取8份球孢链霉菌发酵液,加入6份柠檬酸,再加入7份蟹壳提取物,搅拌25min,再加入10份柠檬烯,3份β-水芹烯,5份石竹烯,在40℃下搅拌2h,得消毒剂。该条件下,各种成分可以充分混合,使消毒剂更加均匀和稳定,提高了产品在实际应用过程中的效果。
本发明的操作步骤中的常规操作为本领域技术人员所熟知,在此不进行赘述。
以上所述的实施例对本发明的技术方案进行了详细说明,应理解的是以上所述仅为本发明的具体实施例,并不用于限制本发明,凡在本发明的原则范围内所做的任何修改、补充或类似方式替代等,均应包含在本发明的保护范围之内。
SEQUENCE LISTING
<110> 浙江省淡水水产研究所
<120> 一种红螯螯虾孵卵繁育方法
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<170> PatentIn version 3.5
<210> 1
<211> 18
<212> PRT
<213> 人工合成
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Claims (9)
1.一种红螯螯虾孵卵繁育方法,包括亲虾培育,脱卵,消毒,孵化,其特征在于:所述消毒过程中所用的消毒剂成分及其重量份为:球孢链霉菌发酵液3-8份,蟹壳提取物4-7份,柠檬烯5-10份,β-水芹烯1-3份,石竹烯2-5份,柠檬酸3-6份。
2.根据权利要求1所述的一种红螯螯虾孵卵繁育方法,其特征在于:所述的球孢链霉菌发酵液的制备方法为:取1-3份球孢链霉菌,加入9-12份无菌生理盐水得菌体悬浮液,向悬浮液中加入4-6份活性短肽,再接种到40-50份的察氏培养液中,置于20-23℃、170-175r/min的恒温摇床上培养4-5d,离心分离,取上清液得链霉菌发酵液。
3.根据权利要求2所述的一种红螯螯虾孵卵繁育方法,其特征在于:所述的球孢链霉菌发酵液的制备过程中所加入的活性短肽的氨基酸序列为:HSHACHPGKLCVCVNCSR。
4.根据权利要求2所述的一种红螯螯虾孵卵繁育方法,其特征在于:所述的察氏培养液的成分及其重量份为:5-7份蔗糖,1-3份FeSO4,1-2份NaNO3,2-3份K2HPO4,1-4份MgSO4·7H2O,5-7份KCl,10-13份蒸馏水。
5.根据权利要求1所述的一种红螯螯虾孵卵繁育方法,其特征在于:所述的蟹壳提取物的制备方法为:取10-15份蟹壳,用10-13份质量分数为12%-15%HCl溶液浸泡5-7h,过滤,水洗至中性,置于8-10份温度为质量分数为35%-40%NaOH溶液中,在温度为96-100℃下搅拌15-17h,离心分离,将所得固体水洗至中性,得蟹壳提取物。
6.根据权利要求1所述的一种红螯螯虾孵卵繁育方法,其特征在于:所述的消毒剂的制备方法为:取3-8份球孢链霉菌发酵液,加入3-6份柠檬酸,再加入4-7份蟹壳提取物,搅拌20-25min,再加入5-10份柠檬烯,1-3份β-水芹烯,2-5份石竹烯,在35-40℃下搅拌1.5-2h,得一种红螯螯虾孵卵繁育方法。
7.根据权利要求1所述的一种红螯螯虾孵卵繁育方法,其特征在于:所述的消毒过程为:取1-3份消毒剂,加入40-45份水,搅拌混合得消毒液,取2-5份虾卵,加入制备的消毒液,浸泡3-6h,取出。
8.根据权利要求1所述的一种红螯螯虾孵卵繁育方法,其特征在于:所述的亲虾培育过程为:亲虾按雌雄比例3-4∶1进行专池培育;所述的脱卵过程为:采用消毒后的镊子将虾卵从亲虾腹部分离。
9.根据权利要求1所述的一种红螯螯虾孵卵繁育方法,其特征在于:所述的孵化过程为:将消毒后的卵虾置于孵化器中,孵化密度为125-130粒/mL,孵化池内溶解氧保持在10-13mg/L,孵化温度保持在25-27℃,孵化液由水、水解蛋白、α-亚麻酸、复合维生素、葡萄糖、葡萄糖酸钙、硫酸锌、海藻酸钠按100/4/0.3/0.4/7/4/0.2/7的重量比混合制成。
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