【作者】 王品虹；

【导师】 常亚青；

【作者基本信息】 中国科学院研究生院（海洋研究所） ， 海洋生物学， 2007， 博士

【摘要】 本论文以大连沿海养殖刺参劾皱纹盘鲍为研究对象,利用组织学、超微病理学、病理生理学的方法,开展了患病病理学研究,并对可能诱发刺参和皱纹盘鲍病害的原因,以及病变过程进行了初步探讨。中国北方养殖刺参[Apostichopus japonicus (Selenka)]幼参和成参的急性口围肿胀病。急性口围肿胀病是爆发于中国北方辽宁省和山东省沿岸养殖刺参群体中的一种新发现的疾病,并且从2004年开始已经引起刺参的大面积死亡现象,经济损失重大。本文报道在养殖患病刺参的肠上皮细胞内发现大量的病毒样颗粒。这是首次报道病毒样粒子感染刺参。组织学检查表明该病毒具有包涵体结构,寄生于肠上皮细胞中。电镜检查的结果表明,该病毒粒子呈球形,直径80-100纳米,螺旋状核衣壳,具有囊膜结构且囊膜表面具有纤突结构。进一步的形态学和病理学分析发现该病毒具有许多报道的关于冠状病毒的特征。细胞质内的病毒颗粒大部分以团聚方式存在于一个完整的包膜内,形成典型的病毒包涵体结构。最明显的细胞病理学特征是细胞质内大面积的粒状物质的存在,该区域相对缺少相应的细胞器。在病毒包涵体内的管状结构,核衣壳包涵体以及双层膜囊泡也在病变细胞内发现。在病参体内未发现立克次氏体,衣原体,细菌以及其他寄生生物。中国北方养殖皱纹盘鲍(Haliotis discus hannai)的肌肉萎缩症病理研究。在2004年和2005年的中国北方海区,养殖皱纹盘鲍幼体和成体群体中爆发了严重的肌肉萎缩症。肌电图监测显示病鲍的肌肉萎缩是肌源性的而不是神经源性的。与正常对照组对比结果显示,病鲍的肌纤维数量和肌纤维直径都显著降低。肌酶谱测定结果显示,病鲍血清中的肌酸激酶水平,肌酸激酶—肌同工酶水平和乳酸脱氢酶水平,与正常血清对比,呈显著升高状态,说明病鲍体内发生了肌细胞损伤。超薄切片电镜检测结果显示在发生病变的肌细胞内存在双螺旋丝状结构的包涵体(Paired helical filaments, PHFs),该结构是人类患包涵体肌炎的主要的病理特征。本文首次报道在无脊椎动物中发现双螺旋丝包涵体结构。对损伤的肌纤维的进一步研究发现了异常增生的小圆柱体结构(Small Cylinder Structure, SCS)和致密小体(Dense colored particles, DCP),同时观察到这两种异常结构与肌纤维的损伤直接相关。本文还对该病的感染机制作了探讨。中国北方皱纹盘鲍(Haliotis discus hannai)肌肉萎缩的病理生理研究。严重的足肌肉萎缩症是鲍的一种慢性致死性疾病。在中国北方的养殖皱纹盘鲍群体中第一次发现是在2000年,随后在各养殖海区,自然海区及实验室养殖过程中都发现了该病的暴发。超微结构电镜检测显示肌肉纤维损伤严重,其内的大部分肌原纤维断裂或消失。大量的花瓣状的糖原颗粒聚集在断裂的肌原纤维中,还有一些被包裹在包膜内。这说明在病鲍体内的糖原代谢处于被抑制状态(或停止状态),随后的血清学检测表明病鲍血清中的葡萄糖含量与正常对照的含量显著降低,也证明了糖代谢途径的终止。病鲍血清中胆固醇和甘油三酯的量显著降低,伴随着高密度脂蛋白的水平显著升高,说明脂肪代谢途径的亢进状态。病鲍血清中的二氧化碳水平的显著降低说明出现了酸碱平衡紊乱,通过病因学分析可知该酸碱平衡紊乱应属于代谢性酸中毒。病鲍血清中的钠离子和钾离子浓度显著升高,无机磷和镁离子浓度显著降低说明出现了患病皱纹盘鲍体内出现了电解质紊乱。病鲍血清中蛋白质含量的检测表明患病皱纹盘鲍已出现低蛋白血症,说明病鲍出现了严重的营养不良。病鲍血清中的尿酸含量,胆碱酯酶水平和γ-转肽酶水平与正常对照组相比显著降低。各项指标检测的结果说明糖代谢途径的终止造成了病鲍一系列的调节体系的不平衡和各种生理功能的紊乱,甚至造成了鲍的最终死亡。更多还原

【Abstract】 Acute peristome edema disease in juvenile and adult sea cucumbers [Apostichopus japonicus (Selenka)] reared in North China. Acute peristome edema disease (APED) is a new disease that broke out in cultured sea cucumber along the Shangdong and Liaoning province coasts in China, PR, and has caused a great deal of death in Apostichopus japonicus(Selenka)since 2004. Here we report virus-like particles found in intestine epithelium of sea cucumbers reared in North China. It is the first time that sea cucumbers are reported to be infected by virus. Histological examinations showed that the viral inclusion bodies existed in intestine epithelium cells. Electron microscopic examinations show that the virions were spherical, 80—100 nm in diameter, and composed of a helical nucleocapsid within an envelope with surface projections. Detailed studies on the morphogenesis of these viruses found many characteristics previously described for coronaviruses. Virus particles always congregated, and formed a virus vesicle with an encircling membrane. The most obvious cellular pathologic feature is large granular areas of cytoplasm, relatively devoid of organelles. Tubular structures within virus-containing vesicles, nucleocapsid inclusions, and double-membrane vesicles are also found in the cytopathic cells. No rickettsia, chlamydia, bacteria, or other parasitic organisms were found.Pathology of atrophic muscle in abalones (Haliotis discus hannai) reared in North China. Severe muscle atrophy disease in cultured abalone juveniles and adults were reported in 2004 and 2005 in North China. Electromyography test shows that the atrophy of abalones is myogenetic but not neurogenetic. Comparing to the control group, the number of intact myofibrils and the diameter of myofibrils of the diseased ones significantly decreased statistically. The levels of Creatine Kinase (CK), Creatine kinase—MM(CK--MM) and Lactate Dehydrogenase (LDH) in serum of diseased abalones increased significantly comparing to the control, which suggests that damage of muscle cells occurred. Ultrathin-section electron microscopic examination was performed and paired helical filaments (PHFs) inclusion bodies were found in cytopathic cells of muscle, which is the pathologic feature reported for atrophic muscles of human patients with inclusion body myositis. It is the first time that PHFs inclusion bodies were reported found in invertebrate. Further research on myofibril damages demonstrated that two abnormal structures, Small Cylinder Structure (SCS) and Dense-Colored Particles (DCPs), proliferated in broken myofibrils, which were observed to be related directly to myofibril damage. The mechanism of the infection was discussed.Pathophysiology of muscle atrophy in abalones (Haliotis discus hanni) reared in China. Severe amyotrophy syndrome of cultured abalone was a chronic disease and the disease breaks out in North China in May every year. Ultrathin-section electron microscopic examination was performed and myofibers were observed to be damaged severely, within which most of myofibrils were broken. Large numbers of petaloid glycogen particles, some of which were encircled in membranes, were observed accumulated among broken myofibrils. It suggests that the glucose metabolism were inhibited in the disease, which is proved by the low level glucose detected in blood. Lower levels of cholesterol and triglyceride, together with high level of high-density lipoprotein in blood suggest the promotion of lipolysis. Low levels of total carbon dioxide in blood suggest acid-base disturbance, which belongs to metabolic acidosis by its etiologic analyses. Electrolytes imbalances were characterized by low levels of sodium and potassium accompanied with high levels of phosphate and magnesium in blood. Hypoproteinemia also appeared in diseased abalones, which suggest the diseased abalones were in severe dystrophia. The content uric acid and the levels of choline esterase andγ-transpeptidase in blood all decreased sharply compared to the control. It can be concluded from the results that the inhibition of glucose metabolism brings series of imbalances and disturbances to the physiological functions of the abalones, and even caused them to death.更多还原