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.

静息卵形成机制收集的20个轮虫菌株中，有8个菌株具有有性繁殖能力。HPLC分析表明，在30℃和16 ppt盐度下，静息卵形成旺盛。 γ-氨基丁酸和 5-HT 存在于轮虫中，表明它们可能在轮虫中充当神经递质。从Koshiki株中获得的雄性配偶识别信息素多克隆抗体，除了Hamana株和西班牙株外，与雌性株结合时，雄性配偶识别同样受到抑制。这表明其他小株的污染。 -物种轮虫菌株不会降低群体中雄性配偶识别度。休眠卵的大规模生产和保存使用夏威夷和哈马纳菌株，我们在上述获得的最佳条件下进行了休眠卵的大量生产。采用半连续培养法，增强了较长时间的混合诱导，对于夏威夷品系，每500升罐中三个罐的静息卵收获量分别为7、5和1.1（百万）个，分别为65、79个。 Hamana品系的静息卵2700万枚，冻干后在4℃下大量保存，孵化率达到50-60%。