首页> 外国专利> METHOD FOR PRODUCING FREE SWIMMING ARTEMIA NAUPLII AND PACKAGED CYSTS FOR USE IN THAT METHOD

METHOD FOR PRODUCING FREE SWIMMING ARTEMIA NAUPLII AND PACKAGED CYSTS FOR USE IN THAT METHOD

机译:制备游离游动性青蒿素和包装的半胱氨酸的方法

摘要

1. A method for producing free swimming Artemia nauplii starting from a quantity of Artemia cysts, including diapauzing cysts, by incubating the cysts in a hatching medium, in a density which comprises maximally about 5 g cyst dry matter per litre hatching medium, for a predetermined incubation period and under conditions allowing at least a portion of the cysts to hatch and to release free swimming nauplii within this predetermined incubation period, in which method the cysts are brought in contact with hydrogen peroxide to break diapause in at least a number of said diapauzing cysts in order to enable them to produce free swimming nauplii within said predetermined incubation period, characterised in that said hydrogen peroxide is introduced in the hatching medium in an amount selected within the range between 0.5 and 30mg/l hatching medium. 2. A method according to claim 1, characterised in that the selected amount of hydrogen peroxide is lower than 20mg/l and more particularly lower than 15mg/I and is higher than 1mg/l, in particular higher than 2mg/l and more particularly higher than 3mg/l. 3. A method according to claim 1 or 2, characterised in that the cysts are hatched in said hatching medium in a density of less than 4 g cyst dry matter per litre, in particular in a density of less than 3 g cyst dry matter per litre and preferably in a density of about 2 g or less cyst dry matter per litre. 4. A method according to any one of the claims 1 to 3, characterised in that said cysts comprise such an amount of diapauzing cysts that by introducing an optimal amount of hydrogen peroxide in the hatching medium, the hatching percentage can be increased from X % to a maximum hatching percentage of M % and said amount of hydrogen peroxide is selected within said range to increase the hatching percentage from X % to a hatching percentage which is at least equal to X % + 0.4* (M-X) %, and preferably to a hatching percentage which is at least equal to X % + 0.6* (M-X) %. 5. A method according to claim 4, characterised in that said maximum hatching percentage is determined by taking a number of samples of the cysts, by hatching these samples in hatching media differing from one another by a different amount of hydrogen peroxide introduced per volume unit therein and situated within said range, by determining the hatching percentages for the different hydrogen peroxide amounts, and by selecting the highest hatching percentage amongst the determined hatching percentages. 6. A method according to any one of the claims 1 to 3, characterised in that for selecting said amount of hydrogen peroxide, a number of samples of the cysts are taken, these samples are hatched in hatching media differing from one another by a different amount of hydrogen peroxide introduced per volume unit therein and situated within said range, the hatching percentages are determined for the different hydrogen peroxide amounts and the maximum percentage M % amongst these hatching percentages, and a hydrogen peroxide amount is selected which increases the hatching percentage from X % to a value higher than X %, in particular a hydrogen peroxide amount which gives either the maximum hatching percentage of M % or a hatching percentage which is at least equal to X % +0.4* (M-X) % and preferably at least equal to X % + 0.6* (M-X) %, a hydrogen peroxide amount corresponding substantially to said hydrogen peroxide amount which gives said maximum hatching percentage of M % being most preferably selected. 7. A method according to any one of the claims 1 to 6, characterised in that said hydrogen peroxide is introduced in the hatching medium by adding hydrogen peroxide and/or at least one compound to this medium which produces hydrogen peroxide in the hatching medium, the hydrogen peroxide being preferably introduced in the hatching medium by adding at least one hydrogen peroxide producing compound which contains a peroxide link and which is selected more particularly from the group comprising peroxides, perborates, persulfates, peracetates or combinations thereof to the hatching medium, the hydrogen peroxide producing compound being preferably a solid compound and comprises most preferably calcium and/or magnesium peroxide. 8. A method according to any one of the claims 1 to 7, characterised in that said hydrogen peroxide is introduced in the hatching medium less than 10 hours, preferably less than 8 hours, more preferably less than 6 hours and most preferably less than 5 hours after having started to incubate the cysts therein. 9. A method according to claim 8, characterised in that said hydrogen peroxide is introduced substantially simultaneously with the cysts in the hatching medium, in particular by being mixed with the cysts and/or by being adhered or coated thereon. 10. A method according to any one of the claims 1 to 9, characterised in that said predetermined incubation period is a period of about 24 hours or less and at least 90 % of the cysts which hatch within an incubation period of 48 hours are hatched within this predetermined incubation period. 11. Artemia cysts destined to be incubated in a hatching medium to produce free swimming nauplii, which cyst show a hatching percentage of X% of the total number of full cysts when they are hatched in the hatching medium in the absence of hydrogen peroxide, characterised in that the Artemia cysts are combined with a quantity of hydrogen peroxide and/or of at least one compound which produces hydrogen peroxide when added to the hatching medium, which quantity is sufficient to introduce an amount of hydrogen peroxide in the hatching medium which increases the hatching percentage of the cysts from X% to a hatching percentage higher than X% when this amount of hydrogen peroxide is introduced in the hatching medium. 12. Artemia cysts according to claim 11, characterised in that said quantity of said hydrogen peroxide and/or of said hydrogen peroxide producing compound is determined in such a manner that when adding this quantity to the hatching medium wherein the cysts are to be incubated, the hatching percentage of the cysts is increased from X% to said hatching percentage higher than X%. 13. Artemia cysts according to claim 11 or 12, characterised in that said quantity of said hydrogen peroxide and/or of said hydrogen peroxide producing compound is determined in such a manner that when adding this quantity to the hatching medium wherein the cysts are to be incubated, an amount of hydrogen peroxide comprised between 0.5 and 30mg/l is introduced therein. 14. Artemia cysts according to any one of the claims 11 to 13, characterised in that they are destined to be incubated in the hatching medium in a predetermined density of about 5 g or less cyst dry matter per litre. 15. Artemia cysts according to any one of the claims 11 to 14, characterised in that said cysts comprise such an amount of diapauzing cysts that by introducing an optimal amount of hydrogen peroxide in the hatching medium, the hatching percentage can be increased from X % to a maximum hatching percentage of M % and said amount of hydrogen peroxide is such that when the cysts are hatched in the presence thereof the hatching percentage is at least equal to X % + 0.4* (M-X) %, and preferably at least equal to X % + 0.6* (M-X) %. 16. Artemia cysts according to any one of the claims 11 to 15, characterised in that the Artemia cysts are combined with a quantity of at least one compound which produces hydrogen peroxide when added to the hatching medium, which hydrogen peroxide producing compound containing preferably a peroxide link and is more particularly selected from the group comprising peroxides, perborates, persulfates, peracetates or combinations thereof, the hydrogen peroxide producing compound being preferably a solid compound and comprises most preferably calcium and/or magnesium peroxide. 17. Artemia cysts according to any one of the claims 11 to 16, characterised in that said quantity of said hydrogen peroxide and/or of said compound comprises and/or produces less than about 15 mg and more than about 0.5 mg, preferably more than about 1 mg and most preferably more than about 1.5 mg hydrogen peroxide per gram cyst dry matter, which quantity is preferably mixed with the cysts and/or adhered or coated thereon. 18. Artemia cysts according to claim 17, characterised in that said quantity of said hydrogen peroxide and/or of said compound comprises and/or produces less than about10 mg and preferably less than about 7.5 mg hydrogen peroxide per gram cyst dry matter. 19. Artemia cysts according to any one of the claims 11 to 18, characterised in that the cysts are jointly packaged with said quantity of hydrogen peroxide and/or of at least one compound which produces hydrogen peroxide when added to the hatching medium. 20. Artemia cysts according to claim 19, characterised in that said quantity of said hydrogen peroxide and/or of said hydrogen peroxide producing compound is contained in a further packaging separating this hydrogen peroxide and/or this compound from the cysts. 21. Artemia cysts according to claim 19 or 20, characterised in that said quantity of said hydrogen peroxide and/or of said compound is contained in the same packaging as the cysts and is in particular mixed with the cysts and/or adhered or coated thereto.
机译:1.一种生产自由游泳卤虫无节幼体的方法,该方法从一定数量的卤虫囊肿开始,包括滞育的囊肿,通过在孵化介质中孵育囊肿,其密度最大为每升孵化介质包含约5 g囊肿干物质。在预定的潜伏期中,并且在允许至少一部分囊肿在该预定的潜伏期中孵化并释放出自由游泳无节幼体的条件下,在这种方法中,至少将多个所述囊肿与过氧化氢接触以破坏滞育为了使它们能在所述预定的温育期内产生自由游泳的无节幼体,使卵囊悬浮,其特征在于将所述过氧化氢以0.5至30mg / l的孵化培养基中的量引入到孵化培养基中。 2.根据权利要求1所述的方法,其特征在于,所选择的过氧化氢的量低于20mg / l,更特别地低于15mg / l,并​​且高于1mg / l,特别是高于2mg / l,并​​且更特别地高于3mg / l。 3.根据权利要求1或2所述的方法,其特征在于,在所述孵化介质中以小于每升4g的孢子干物质的密度,特别是小于小于3g的孢子干物质的密度进行孵化。升,优选密度为每升约2 g或更少的囊肿干物质。 4.根据权利要求1至3中任一项所述的方法,其特征在于,所述囊肿包括一定数量的悬浮的囊肿,使得通过在孵化介质中引入最佳量的过氧化氢,可以将孵化百分比从X%增加。到最大孵化百分比为M%,并且在所述范围内选择所述过氧化氢的量,以将孵化百分比从X%增加到至少等于X%+ 0.4 *(MX)%的孵化百分比,并且优选为孵化百分比至少等于X%+ 0.6 *(MX)%。 5.根据权利要求4所述的方法,其特征在于,所述最大孵化率是通过采集多个囊肿样品,通过在彼此不同的孵化介质中孵化这些样品而确定的,所述孵化介质彼此不同,每体积单位引入的过氧化氢量不同通过确定不同过氧化氢量的孵化百分比,并在所确定的孵化百分比中选择最高的孵化百分比,将其置于所述范围内。 6.根据权利要求1至3中任一项所述的方法,其特征在于,为了选择所述量的过氧化氢,采集多个囊肿样品,这些样品在彼此不同的孵化介质中孵化。每单位体积引入的过氧化氢的量并处于所述范围内,针对不同的过氧化氢量确定孵化百分比,并在这些孵化百分比中确定最大百分比M%,并选择增加孵化百分比的过氧化氢。 X%到高于X%的值,特别是过氧化氢的量,该过氧化氢的量等于或大于M%的最大孵化百分比,或者至少等于X%+ 0.4 *(MX)%的孵化百分比,优选至少等于至X%+ 0.6 *(MX)%,最优选基本上对应于给出所述最大孵化率M%的所述过氧化氢量的过氧化氢量。精挑细选。 7.根据权利要求1至6中任一项所述的方法,其特征在于,通过将过氧化氢和/或至少一种化合物添加到该介质中在孵化介质中产生过氧化氢来将所述过氧化氢引入孵化介质中,优选通过向孵化介质中加入至少一种包含过氧化物连接并且更特别选自过氧化物,过硼酸盐,过硫酸盐,过乙酸盐或它们的组合的产生过氧化氢的化合物,将过氧化氢引入到孵化介质中,产生过氧化氢的化合物优选为固体化合物,并且最优选包含过氧化钙和/或镁。 8.根据权利要求1至7中任一项所述的方法,其特征在于,将所述过氧化氢在小于10小时,优选小于8小时,更优选小于6小时且最优选小于5小时的时间内引入孵化介质中。开始在其中培养囊肿后数小时。 9.根据权利要求8所述的方法,其特征在于,所述过氧化氢与所述囊肿基本上同时在孵化介质中被引入,特别是通过与所述囊肿混合和/或通过粘附或包被在其上。 10.根据权利要求1至9中的任一项所述的方法,其特征在于,所述预定的温育期为约24小时或更短的时间,并且在该预定的温育期内孵化在48小时的温育期内孵化的囊肿的至少90%。 11.注定要在孵化培养基中孵育以产生自由游泳无节幼体的卤虫囊肿,当它们在没有过氧化氢的孵化培养基中孵化时,其孵化率占完整囊肿总数的X%。因为卤虫囊肿与一定量的过氧化氢和/或至少一种化合物混合在一起,当添加到孵化介质中时会产生过氧化氢,该量足以在孵化介质中引入一定量的过氧化氢,从而增加孵化率。当在孵化培养基中引入过氧化氢时,囊肿的孵化百分比从X%到高于X%的孵化百分比。 12.根据权利要求11所述的卤虫囊肿,其特征在于,所述过氧化氢和/或所述产生过氧化氢的化合物的量以这样的方式确定,即当将该量添加到要培养所述囊肿的孵化介质中时,囊肿的孵化百分比从X%增加到高于X%的孵化百分比。 13.根据权利要求11或12所述的卤虫囊肿,其特征在于,所述过氧化氢和/或所述产生过氧化氢的化合物的量以这样的方式确定,即当将该量添加到其中要形成囊肿的孵化介质中时,温育后,向其中引入介于0.5和30mg / l之间的过氧化氢。 14.根据权利要求11至13中任一项所述的卤虫囊肿,其特征在于,它们预定以每升约5g或更少的囊干物质的预定密度在孵化培养基中孵育。 15.根据权利要求11至14中任一项所述的卤虫囊肿,其特征在于,所述囊肿包含一定量的悬浮的囊肿,使得通过在孵化介质中引入最佳量的过氧化氢,可以将孵化百分比从X%增加。到最大孵化百分比为M%,并且所说的过氧化氢的量是这样的:当囊肿在存在时孵化时,孵化百分比至少等于X%+ 0.4 *(MX)%,优选至少等于X%+ 0.6 *(MX)%。 16.根据权利要求11至15中任一项所述的卤虫囊肿,其特征在于,所述卤虫囊肿与一定量的至少一种当添加到孵化培养基中时产生过氧化氢的化合物结合,所述产生过氧化氢的化合物优选地包含过氧化氢键,更特别地选自过氧化物,过硼酸盐,过硫酸盐,过乙酸盐或它们的组合,产生过氧化氢的化合物优选为固体化合物,并且最优选包含钙和/或镁的过氧化物。 17.根据权利要求11至16中的任一项所述的卤虫囊肿,其特征在于,所述量的所述过氧化氢和/或所述化合物包括和/或产生小于约15mg且大于约0.5mg,优选大于9mg。每克囊肿干物质约1mg,最优选大于约1.5mg过氧化氢,该量优选与囊肿混合和/或粘附或包被在其上。 18.根据权利要求17所述的卤虫囊肿,其特征在于,所述每克囊干物质的所述过氧化氢和/或所述化合物的量包括和/或产生小于约10mg,优选小于约7.5mg的过氧化氢。 19.根据权利要求11至18中任一项所述的卤虫囊肿,其特征在于,所述囊肿与所述量的过氧化氢和/或当添加到孵化介质中时产生过氧化氢的至少一种化合物共同包装。 20.根据权利要求19所述的卤虫囊肿,其特征在于,所述一定量的所述过氧化氢和/或所述产生过氧化氢的化合物容纳在将所述过氧化氢和/或所述化合物与所述囊肿分开的另一包装中。 21.根据权利要求19或20所述的卤虫囊肿,其特征在于,所述一定量的所述过氧化氢和/或所述化合物被包含在与所述囊肿相同的包装中,并且特别地与所述囊肿混合和/或粘附或包被于其上。 。

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