热带海洋珍珠贝类立克次体病研究：Ⅴ.超微病理学及致病机理 …

对海水养殖大珠母贝和合浦珠母贝类立克次体（Ｒｉｃｋｅｔｔｓｉａ－ｌｉｋｅ ｏｒｇａｎｉｓｍ,ＲＬＯ）病的超微病理学进行了研究。结果显示细胞及亚细胞结构经历由变性发展到坏死的病变过程,如胞核由核浓缩,核碎裂发展为核溶解;线粒体出现肿胀,嵴和基质改变及内质网脱颗粒,扩张和囊泡变等一系列混浊肿胀和水泡变性的基本特征;最终,细胞器完全溶解消失使胸质疏松,呈崩解或溶解状。     

热带海洋珍珠贝类立克次体病研究 Ⅳ.组织细胞病理学研究

类立克次体(RLO)对大珠母贝和合浦珠母贝具有强烈的致病性,引起基本一致的病理变化.在急性坏死破坏期内,RLO导致外套膜、鳃、消化管、肝胰腺、生殖腺腺管及全身血管内皮系统等多器官组织的变性坏死,使器官组织结构的完整性遭到破坏,破坏程度与RLO包涵体的数目密切相关,而细胞的破坏与细胞内RLO的生长发育及繁殖到大量数目密切相关.鉴于此,将RLO引起的珍珠贝病称为类立克次体病(RLO病).RLO病呈急性变质性炎症和慢性增生性炎症病理,在前者细胞呈现崩解性坏死、溶解性坏死和退变性坏死;在后者存在实质细胞增生和纤维母细胞增生为纤维细胞并形成纤维化.根据炎症病理的发展过程,RLO病可分为急性坏死破坏期和慢性增生修复期.     

热带海洋珍珠贝类立克次体病研究 Ⅳ.组织细胞病理学研究

类立克次体（RLO）对大珠母贝和合浦珠母贝具有强烈的致病性,引起基本一致的病理变化．在急性坏死破坏期内,RLO导致外套膜、鳃、消化管、肝胰腺、生殖腺腺管及全身血管内皮系统等多器官组织的变性坏死,使器官组织结构的完整性遭到破坏,破坏程度与RLO包涵体的数目密切相关,而细胞的破坏与细胞内RLO的生长发育及繁殖到大量数目密切相关．鉴于此,将RLO引起的珍珠贝病称为类立克次体病（RLO病）．RLO病呈急性变质性炎症和慢性增生性炎症病理,在前者细胞呈现崩解性坏死、溶解性坏死和退变性坏死;在后者存在实质细胞增生和纤维母细胞增生为纤维细胞并形成纤维化．根据炎症病理的发展过程,RLO病可分为急性坏死破坏期和慢性增生修复期．     

大黄鱼溃疡病的组织病理和超微病理研究

利用组织切片及超薄切片电镜技术对患溃疡病大黄鱼的骨骼肌、鳃、肝、肾、脾、肠6种组织进行病理学分析,探讨溃疡病的发生机理.组织病理显示,溃疡病的病变性质为变质性炎症,主要表现为骨骼肌肌丝断裂,炎性浸润;鳃丝肿胀,鳃小片溶融,细胞变性坏死;肝实质细胞空泡化,结缔组织异常;肾脏弥散性出血,肾小管上皮细胞异常;脾脏和肠也产生炎性症状.超微病理显示,骨骼肌肌丝排列紊乱,肌丝碎裂,线粒体肿胀,嵴断裂;肝脏组织及细胞内大量脂肪积聚表现明显的脂肪变性,肝内细胞大量坏死并溶融,核肿胀破裂溶解,胞内线粒体肿胀,嵴断裂;肾脏组织细胞发生形变,胞核异常;脾内细胞可见明显的纤维化;肠壁黏膜细胞结构破坏或消解,肠壁结构严重破坏.结果表明,大黄鱼溃疡病的组织细胞结构发生了明显的病理变化,全身性组织严重损害,肝、脾、肾等重要器官功能衰竭,丧失正常的生理代谢作用;病鱼的摄食能力也逐渐减退或丧失,终至死亡.最后提出一些防治大黄鱼溃疡病的建议.     

斑节对虾 弧菌病的病理学研究

对患有典型弧菌病症状的养殖斑节对虾进行了病理学研究，取病虾血淋巴制作血涂片，并取鳃、肝胰腺、肌肉等组织作组织病理和超微病理观察，同时取肌注感染后的患病对虾测定血淋巴中超氧化物歧化酶（SOD）的活力，结果显示，患病对虾的血细胞数目急剧减少，胞质浓染，透明细胞很难找到，血细胞形态多样，有的伸出伪足或形成异样突起，患病对虾锶、肝胰腺、心脏、肌肉以及胃肠等组织内均出现程度不同的炎症反应，以肝胰腺最为严重，组织中充满大量的浓染血细胞，细胞肿胀，细胞核核质边聚，线粒体肿胀，变性、严重时整个细胞呈溶解状态，此外，在鳃和肝胰腺组织中还发现溶酶体增多的现象，肌注感染2h后，感染对虾的SOD活力即开始显著下降，之后缓慢回升，至48h恢复大部分活力。     

三疣梭子蟹(Portunus trituberculatus)乳化病的组织病理和超微病理研究

利用组织切片及超薄切片电镜技术对患“乳化病”三疣梭子蟹的鳃、肝胰腺、步足肌肉、心肌和性腺5种组织进行病理学分析。组织病理显示:乳化病的病变性质为变质性炎症,主要病变部位在肝胰腺,其病灶表现为水变性、脂肪变性和细胞坏死等症状,步足肌和心肌表现出肌纤维断裂、浑浊等症状。超微病理显示:在肝胰腺和步足肌组织中有大量细菌,肝胰腺细胞结构损伤主要表现为微绒毛损伤脱落、线粒体坏死、溶酶体增多及脂肪滴变性,肌纤维断裂、溶解,血隙中血淋巴细胞受严重侵染。     

云纹石斑鱼淋巴囊肿病的光镜和电镜研究

本文用光镜和电镜对海水网箱养殖患淋巴囊肿病的云纹石斑鱼(Epinethelus moara)进行了研究.分别观察了其囊肿物、皮肤、肝脏、脾脏、肾脏、心脏和肠管等器官.光镜下,可见囊肿物主要由巨大的囊肿细胞构成,这些细胞的细胞质内含有许多包涵体和空泡;肝脏汇管区炎性细胞浸润、肝细胞肿胀并有胞质嗜碱性小体出现;脾脏内白细胞滞留;肾小管变性、萎缩、坏死并形成大面积无肾小管区;心肌炎和心外膜炎等病理变化.电镜下,可见囊肿细胞含有大量的病毒颗粒,这些病毒颗粒散布于核周区域;病毒颗粒切面多呈6角形,核衣壳外大部分还具有一中等电子密度的囊膜,衣壳由3层构成,核髓有明暗相间的区带.本文还对该病与国外报道有差异的问题、病毒的形态和病鱼慢性死亡的病理学等方面作了讨论.     

中华绒螯蟹颤抖病组织病理学研究

采用常规石蜡切片技术、以正常样品作对照,对患颤抖病的中华绒整蟹不同组织进行病理学观察,旨在为颤抖病病原的确定及诊断等提供参考,同时期望加深对河蟹发病机制及死亡过程的认识。结果表明,患病河蟹的鳃、心脏、肝胰腺、腹节神经等组织器官发生了不同程度的病变,其病理特征主要表现为：细胞浊肿、变性、坏死,某些细胞的细胞核固缩、碎裂或崩解;结合前人的研究,推测鳃、心脏、肝胰腺、腹节神经是病原体感染河蟹的主要靶组织器官;河蟹具有重要生理功能的鳃、心脏、肝胰腺组织发生了严重病变,使其功能遭到破坏,这可能是导致河蟹发病死亡的病理学基础。     

对虾白斑杆状病毒(WSBV)对斑节对虾的实验感染与病理学研究

用取自发病虾塘患典型白斑病的病虾为毒源 ,对健康斑节对虾Penaeusmonodon进行人工感染 ,按Koch氏法证实了对虾白斑杆状病毒 (WSBV)是白斑病的病原。对人工感染WSBV后患典型白斑病的斑节对虾的胃、肠及皮下结缔组织进行石蜡切片和超薄切片 ,研究WSBV引起的组织病理变化和宿主细胞感染WSBV后的超微病变过程。结果表明 :感染WSBV后病虾的不同组织均存在广泛的变性、坏死 ,上皮细胞大量解体、脱落 ;部分细胞核肿大、游离 ;结缔组织糜烂 ,呈现空泡化 ;未发现包涵体。透射电镜照片显示 :感染WSBV后的病虾在感染前期 ,靶细胞核膨胀 ,核仁消失 ,核外膜破损 ,染色质靠边分布 ,内质网断裂 ,核糖体脱落 ,线粒体肿胀 ,高尔基体萎缩 ,溶酶体空泡化 ;感染中期 ,病毒粒子在细胞核内大量复制 ,直至充满整个核 ,细胞核内无结构化 ,核膜及质膜皱缩 ;感染后期 ,核胀破 ,病毒粒子从核内释放 ,细胞核呈空囊状 ,胞质内髓样结构明显 ,细胞器解体 ,细胞功能丧失。     

泥螺肝胰腺细胞的超微结构和功能

利用透射电镜技术研究了泥螺(Bullactaexarata)肝胰腺细胞的超微结构。结果表明,泥螺的肝胰腺主要由4类细胞构成:泡状细胞(B细胞)、吸收细胞(R细胞)、纤维细胞(F细胞)和胚细胞(E细胞)。B细胞大,长柱状,胞质中有许多消化泡;R细胞的脂滴数量多,体积大,具少量的线粒体;F细胞含有极丰富的粗面内质网、线粒体等结构,但脂滴的数量远远少于R细胞。E细胞为多边形,核大而圆,粗面内质网少,线粒体小而狭长。根据这4种细胞的超微结构特点,指出了泥螺肝胰腺的功能,并探讨了肝胰腺细胞的分化序列。     

Studies on rickettsia-like organism (RLO) disease of tropical marine pearl oyster-Epidemiological investigation of RLO disease in juvenile populations of maricultured Pinctada maxima

The death which occurred in juvenile population was a severe problem in the course ofPinctada maxima artificial culture. With the methods of field investigation, histological study and statistic analyses, the epidemiological study was carried out on the disease and death in juvenile populations of Pinctada maxima in the Xinying Pearl Oyster Mariculture Farm of Lingao County (for the A, B and C batches of cultured juveniles hanged in the sea) and the Xincun Pearl Oyster Mariculture Farm of Lingshui County (for the D batch of cultured juvenile hanged in the pond), Hainan Province from November 1993 to April 1995.The results show that the deaths which occurred in juvenile populations of Pinctada maxima presented an outbreak pattern. The peak of mortality rates, in general, occurred in 4-to 6-month old pearl oyster juveniles, and the mortality rates gradully declined with the extention of pearl oyster age after cul-turing 8 months. The correlation between the mortality rates of juvenile populations and mean     

热带海洋珍珠贝类立克次体(RLO)病研究 Ⅷ.养殖大珠母贝贝苗养成期群体RLO病流行病学研究

大珠母贝人工养殖过程中,幼贝死亡是一个严重的问题。1993年11月至1995年5月,采用现场调查、实验室研究和数理分析相结合的方法,在海南省临高县新盈珍珠贝养殖场自然海区吊养的A,B,C3批养殖贝和陵水新村珍珠贝养殖场养殖池吊养的D批养殖贝的发病和死亡情况进行了流行病学凋查.结果显示,大珠母贝幼贝群体的死亡是以暴发性方式发生,幼贝群体的大批量死亡高峰一般发生在4～6月龄期,8个月以后随着龄期的延长,死亡率显著降低.幼贝群体死亡率与贝体平均体长的关系是4cm以下的贝体死亡率较高,1～3cm阶段处于死亡高峰期内,5cm以上的贝体死亡率显著降低.大珠母贝幼贝群体的死亡率与类立克次体感染(即RLO平均感染严重度指数SI)之间存在显著的相关性,在RLO感染的高峰之后或当中部相应伴随贝群体死亡率的高峰,随着RLO感染的降低,贝群体的死亡率也明显降低。在4批养殖贝群体中,仅在A批养殖贝群体样本中发现有少量纤毛虫寄生(感染率为87.5%,感染强度为3.56个/10倍物镜).这几批幼贝群体的死亡与海水温度、盐度之间均无相关性。     

Rickettsia-like organism infection associated with mass mortalities of blood clam, Tegillarca granosa, in the Yueqing Bay in China

A series of mass mortalities of the cultured blood clam, Tegillarca granosa, occurred in the Yueqing Bay of China from 2005 to 2009. An obligate intracellular prokaryote, designated as rickettsia-like organism (RLO), was frequently found in the moribund or dead blood clam sample during ultrastructural examination. These organisms were usually round, ellipsoid or occasionally dumbbell-shaped, ranged from approximately 0.28 to 0.71 μm in size and had a trilaminar cell wall. Two reproductive modes of organisms, transverse binary fission and budding, were observed. The organisms were able to form eosinophilic inclusions. Most inclusions were found within epithelial and connective tissues of the mantle, gills and digestive tube. The biological and morphological characteristics indicate that these organisms may belong to the family Rickettsiaceae. RLOs exhibited significant pathogenicity. Cytopathological examinations revealed extensive necrosis and destruction in the infected cell. The degree of tissue destruction was positively related to the number of RLO inclusions in the tissues, and the cytopathological effects were positively related to the number of intracellular RLO. RLOs and their inclusions were discovered throughout different disease areas and in different time periods. The infection intensity of the RLOs was positively correlated with the mortality rate of clams. Therefore, RLO infection might be associated with mass mortalities of cultured blood clams in the Yueqing Bay.     

Studies on rickettsia-like organism(RLO)disease of tropical marine pearl oyster--Epidemiologicai investigation of RLO disease in juvenile populations of maricultured Pinctada maxima

The death which occurred in juvenile population was a severe problem in the course ofPinctada maxima artificial culture. With the methods of field investigation, histological study andstatistic analyses, the epidemiological study was carried out on the disease and death in juvenile popula-tions of Pinctada maxima in the Xinying Pearl Oyster Mariculture Farm of Lingao County (for the A,B and C batches of cultured juveniles hanged in the sea) and the Xincun Pearl Oyster Mariculture Farmof Lingshui County (for the D batch of cultured juvenile hanged in the pond), Hainan Province fromNovember 1993 to April 1995. The results show that the deaths which occurred in juvenile populations of Pinctada maxima pre-sented an outbreak pattern. The peak of mortality rates, in general, occurred in 4-to 6-month old pearloyster juveniles, and the mortality rates gradully declined with the extention of pearl oyster age after cul-turing 8 months. The correlation between the mortality rates of juvenile populations and mean bodylengths of juvenile populations show that the mortality rate become higher under 4 cm of mean bodylengths of juvenile population and become obvious declined over 5 cm of mean body lengths. The peak ofmortality rate occurred in 1 ～3 cm of mean body lengths. The results of histological observation showedthat rickettsia-like organism inclusions were common, histological widespread infective agent among ev-ery batches (A, B, C and D) of cultured juvenile populations. The mean severity indices (SI) of rick-ettsia-like organism (RLO) infection were positively correlated with mortality rates of juvenile popula-tions. After or within every peaks of RLO infection were all accompanied with the peaks of mortalityrates of host populations and the mortality rates declined with decreased RLO infection. So the evidencesof histological observation and epidemiology in this study indicated that rickettsia-like organism (RLO)may be as an important pathogenic organism of disease and death of cultured Pinctada maxima. In addi-tion, a few ciliates were discovered only in part of batch A of cultured juvenile population. The mortalityrates in juvenile populations were not correlated with the weekly mean temperature and salinity of seawater.     

Studies on rickettsia-like organism (RLO) disease of tropical marine pearl oyster——Epidemiological investigation of RLO disease in larvae populations of maricultured Pinctada maxima

The onset and sPread of diseases in dricultured Anrnals are resulted frOm the mutule ef-fect of pathogen, hOSt and environrnnt. Epidemology takes the diseased AnInal poPulationsas the study object, rnakes observation and exPeriment on the field as the primp method ofstUdy and stuha the mutule effec Of PathOgen, hOS and environrnen by inVeSigating the na-ttire distribution of diseases and ePidendc mechedsm. To date, succeSSful exarnple of epidend-obo study on lerne shellfish deseases is…     

真鲷(Pagrosomus major)肠上皮组织中类立克 次体的超微形态与细胞病理学的初步研究

运用电镜技术对青岛地区患病真鲷进行了病原体检测,在肠上 皮组织细胞中发现一种类立克次体病原体。研究表明,该病原体具有完整的细胞包膜, 内部 具有低电子密度拟核,在胞浆中以两种类型存在-繁殖型和静止型;繁殖型类立克次体呈圆 形、椭圆、哑铃、长棒状,其长轴为0.2～0.8μm,形成包涵体,以分裂方式进行增殖;静止 型类立克次体为圆形,中等电子密度,直径为40～70nm,由繁殖型类立克次体出芽增殖形成 ;线粒体参与了包涵体的形成和类立克次体的增殖过程。细胞病理变化为上皮细胞肿胀并部 分解体,微绒毛破坏,线粒体变形,部分内嵴溶解;内质网膨胀,核糖体脱落。类立克次体 是导致鱼类病害的重要病原体。     

热带海洋珍珠贝类立克次体(RLO)病研究 Ⅶ 大珠母贝育苗期幼虫群体RLO病流行病学研究

对流行病学分析显示大珠母贝幼虫群体的死亡是以暴发性方式发生,从胚胎发育前期至变态期,幼虫存在两个死亡高峰,分别位于壳顶期之前或壳顶期和壳顶后期.大珠母贝幼虫群体的死亡率与RLO感染之间存在显著的相关性,在RLO感染的高峰之后均相应伴随幼虫群体死亡率的高峰,随着RLO感染率和感染强度的降低,幼虫群体死亡率也明显降低.在TEM下未发现未受精卵母细胞、受精卵和胚胎早期幼虫感染RLO,最先出现RLO包涵体阳性感染的时期为24h尚未摄食的D形期幼虫,故认为RLO可能不是通过垂直传播,而是通过外界接触感染的方式感染宿主,贝体感染至发病死亡存在潜伏期.     

合浦珠母贝代谢产物对异枝麒麟菜生长的促进作用

于１９９５年４月间在海南省三亚市、陵水县分别采集合浦珠母贝和异枝麒麟菜并进行混养实验，测定贝释放的三氮（ＮＨ４－Ｎ，ＮＯ３－Ｎ，ＮＯ２－Ｎ）以及藻类对贝释放的三氮的吸收及其生长，在期探讨合浦珠母贝代谢产物促进异枝麒麟菜生长的原因。结果表明，贝释放物中以ＮＨ４－Ｎ为主，ＮＯ３－Ｎ次之，ＮＯ２－Ｎ含量较少；藻对三氮的吸收 依此顺序递减。实验组藻体的生长速率明显高于对照组的，在模拟实验组中，通过加入无机