大口黑鲈外周血细胞显微结构、细胞化学特征及吞噬作用的研究
Microstructure and Cytochemical Characterisation and Phagocytosis of the Peripheral Blood Cells in Micropterus salmoides

中文摘要：通过对大口黑鲈外周血细胞的显微结构及细胞化学染色特征和吞噬作用的研究,为大口黑鲈免疫学研究积累资料。Wright's染色表明:大口黑鲈外周血细胞分为红细胞、粒细胞、单核细胞、淋巴细胞和血栓细胞,其中粒细胞又分为Ⅰ型、Ⅱ型和Ⅲ型。外周血中成熟红细胞最多,占血细胞总数的98.12%,其次为淋巴细胞和血栓细胞,占白细胞中的比例分别为60.92%和22.99%;单核细胞最大,大小为(9.89±0.70) μm×(8.72±0.68) μm;小淋巴细胞最小,大小为(3.88±0.46) μm×(3.48±0.40) μm。大口黑鲈外周血细胞的免疫相关酶ACP、AKP、PO、POX及细胞成分PAS反应、SBB染色的结果表明,所有红细胞的细胞化学染色均呈阴性,不同白细胞的细胞化学染色特征存在差异。其中所有白细胞的AKP、POX染色均呈阴性,PAS反应均呈阳性,除Ⅱ型粒细胞外所有白细胞的PO、ACP染色均呈阳性,除单核细胞外所有白细胞的SBB染色均呈阳性。大口黑鲈外周血细胞吞噬酵母菌的实验表明,红细胞能够吞噬酵母菌,其吞噬率为(15.70±1.07)%,也观察到部分白细胞吞噬酵母的现象。PO和ACP及脂类和糖类可能是大口黑鲈外周血细胞吞噬作用的重要酶类和能量来源。
英文摘要：To provide more information of fish immunology, the study investigated the morphological and cytochemical characterisation and phagocytosis of the peripheral blood cells in Micropterus salmoides. Erythrocyte, granulocyte (type Ⅰ, Ⅱ and Ⅲ), lymphocyte, thrombocyte and monocyte were identified and characterized in blood smear by staining with Wright's. The proportion of erythrocyte occupied the majority (98.12%) of the peripheral blood cells, followed by lymphocyte and thrombocyte, respectively. The size of the monocytes [(9.89±0.70) μm×(8.72±0.68) μm] was the largest, while the small lymphocytes [(3.88±0.46) μm×(3.48±0.40) μm] was the smallest. Blood smears were stained with acid phosphatase (ACP), alkaline phosphatase (AKP), phenoloxidase (PO), peroxidase (POX), periodic acid schiff (PAS), sudan black B (SBB), and examined using bright field microscopy. The results showed that erythrocyte was all negative for the six reactions, and variabe positive reactions with the leukocyte were detected. PAS positivity was observed in all leukocytes, while no AKP and POX positivity was observed. Granulocyte (type Ⅰ and Ⅲ), lymphocyte, thrombocyte and monocyte were positive for PO and ACP activity, while type Ⅱ granulocyte was negative for peroxidase activity. SBB was detected in all leukocytes, except monocytes. In our phagocytosis experiments, (15.70±1.07)% of erythrocyte was able to engulf yeast and part of leukocyte, such as thrombocytes and granulocytes, also was able to engulf yeast. PO and ACP activity and the presence of glycogen and lipid were related to the phagocytic functions of certain cell types requiring important enzyme and energy sources. 2015,34(1): 69-75,80 收稿日期：2014-7-26 DOI：10.3969/j.issn.1000-7083.2015.01.011 分类号：Q246;S942.1 基金项目：农业部热带亚热带水产资源利用与养殖重点实验室开放课题;四川省教育厅重点项目(14ZA0019);"优鲈1号"大口黑鲈在四川地区的养殖推广 *通讯作者：杨淞,E-mail:ysys210@hotmail.com 参考文献： 白俊杰, 李胜杰. 2013. 大口黑鲈遗传育种[M]. 北京: 海洋出版社. 蔡完其, 轩兴荣. 2003. 红鲤4群体间红细胞免疫功能及其差异[J]. 中国水产科学, 10(2): 133-136. 曹伏君, 朱晓燕. 2006. 红鳍笛鲷外周血细胞的显微结构观察[J]. 海洋科学, 30(5): 32-35. 邓国成, 谢骏,