栉孔扇贝血细胞数量变化与生长的关系

A Relationship Between Haemocyte Count and Growth of Scallop Chlamys farreri

于2009年3月至2010年1月期间,每月中旬分别从青岛和威海两地区采集人工养殖和野生的栉孔扇贝(Chla-mys farreri)。采集的扇贝经壳长测量后,于闭壳肌血窦中取血,离心,重悬,超声波破碎后制得血细胞破碎液。从抗栉孔扇贝血细胞的单克隆抗体库中筛选出1株与颗粒血细胞和透明血细胞均能结合,并能与血细胞多个蛋白结合的单克隆抗体作为酶联免疫吸附法(ELISA)中的第一抗体;第二抗体为碱性磷酸酶标记的羊抗鼠抗体。将血细胞破碎液包被于酶标板孔中,经一抗、二抗孵育后,显色读数,分析血细胞数量与生长的关系。结果表明:两地区人工养殖和野生扇贝的壳长均由3月的2 cm左右增长到了次年1月的7 cm以上,其中4～7月增长较快,而8～1月相对缓慢。两地区人工养殖和野生扇贝的血细胞数量于3～6月间均维持在较高的水平,6月达到最高峰,随后急剧下降,并于8或9月达到最低值,此后10～1月有所回升,但仍显著低于3～6月的水平。结论认为:栉孔扇贝血细胞数量与生长的关系在两地区、两扇贝品种间差别不大;但3～6月栉孔扇贝生长较快,血细胞数量相对高;而8～10月栉孔扇贝生长缓慢,其血细胞数量较低。

In this paper,wild and artificial breeding species of scallop Chlamys farreri were collected monthly from two scallop culture areas in Qingdao and Weihai during the period from March 2009 to January 2010.After measurement of shell height,the collected scallops were bled from adductor muscle sinus,and the resulting haemolymph was centrifuged,and haemocyte pellet was collected.Afterwards,the haemocyte pellet was resuspended,sonicated and frozen at-80 ℃.Subsequently,a monoclonal antibody which was against all types of haemocytes in C.farreri,and reacted to many protein in haemocytes was screened out as the primary antibody,in the meantime,alkaline phosphatase labeled goat anti-mouse antibody was employed as the secondary antibody in enzyme-linked immunoadsorbent assay（ELISA）.Thereafter,the collected haemocyte samples were coated in 96-well plates,and incubated successively with primary antibody,secondary antibody and substrate solution,then read at 405 nm.The results showed that shell height of scallops in two species and two areas both increased from about 2 cm in March 2009 to above 7 cm in January 2010.Additionally,shell height grew fast from April to July,while slow from August to January.Haemocyte count of scallops in two species and two areas both maintained a high level from March to June.After June,it decreased dramatically,and reached the minimum in August or September.Afterwards,it recovered gradually from October to January,however,which was still significantly lower than the level in early 2009.In conclusion,seasonal variation of haemocyte count in C.farreri didn＇t present a significant diversity between two species and two areas.Besides,scallops grew fast and had a high haemocyte conut in the period from March to June,while they grew slow and had a low haemocyte conut in the time from August to October.