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BIOLOGICAL ASPECTS OF RESTING EGG FORMATION, DIAPAUSE AND HATCHING OF SMALL SPECIES ROTIFERS AND ITS APPLICATION TO MASS PRESERVATION TECHNOLOGY

小型轮虫静卵形成、滞育和孵化的生物学特征及其在大规模保存技术中的应用

基本信息

批准号：
10660187
负责人：
HAGIWARA Atsushi
金额：
$ 1.41万
依托单位：
NAGASAKI UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10660187/
关键词：
ROTIFERA SMALL SPECIES RESTING EGG PRESERVATION LARVAL REARING Brachionus rotundiformis 休眠卵量産

项目摘要

Mechanism of resting egg formationAmong collected twenty rotifer strains, eight strains had sexual reproductive ability. These exhibited the percent mixis at more than 30% under 30℃ and 16 ppt salinity. Resting egg formation was vigorous with Hawaii and Langkawi strains. The HPLC analysis clarified that gamma-aminobutyric acid and 5- HT are present in rotifers, suggesting that these may function as neurotransmitters in rotifers. We conducted several assays using the polyclonal antibody of male mate recognition pheromone obtained from Koshiki strain. The antibody binded well with strains except Hamana and Spanish strains. Male mate recognition to females among strains were equally inhibited when females were bound with the antibody. These suggest that the contamination of other small-species rotifer strains will not reduce the male mate recognition in the population.Mass production and preservation of resting eggsUsing Hawaii and Hamana strains, we conducted mass production of resting eggs under optimal condition obtained above. Semicontinuous culture method was used to enhance the mixis induction for a longer period of time. For Hawaii strain, the resting egg harvest in three tanks were 7, 5 and 1.1 (million ) resting eggs per 500 liter tank, and it was 65, 79 and 27 million resting eggs for Hamana strain. Resting eggs were mass preserved at 4oC after lyophilization, and we obtained 50-60% hatching.

休眠卵形成机制在收集的20个品系中，有8个品系具有有性生殖能力。在30℃、盐度16 ppt条件下，混合率均在30%以上。夏威夷和兰卡威品系的休眠卵形成旺盛。HPLC分析表明，γ-氨基丁酸和5-羟色胺存在于轮虫中，这表明它们可能作为神经递质在轮虫中发挥作用。我们利用从Koshiki株获得的雄性配偶识别信息素的多克隆抗体进行了几次测定。该抗体与除Hamana和Spanish株外的其它株均能较好地结合。当雌性与抗体结合时，菌株之间的雄性对雌性的配偶识别同样受到抑制。这些结果表明，其它小种轮虫品系的污染不会降低种群中雄虫的配偶识别能力。休眠卵的大量生产和保存利用夏威夷品系和Hamana品系，在上述优化条件下进行了休眠卵的大量生产。采用半连续培养的方法，可以在较长的时间内提高混合物的诱导。对于夏威夷株，每500 L罐的休眠卵收获量分别为7、5和110万枚，而对于Hamana株，每500 L罐的休眠卵收获量分别为65、79和27万枚。冷冻干燥后，将休眠卵大量保存在4 ℃下，我们获得了50-60%的孵化率。