“红肉病”文蛤中发现的一种球形病毒的形态发生与细胞病理学

在患“红肉病”文蛤的消化盲囊上皮细胞中发现了一种球形病毒及其发生基质。病毒粒子呈球形，无囊膜，直径约50－80nm，在细胞质内形成多泡状的包涵体并进行病毒粒子的装配。受感染的细胞具有明显的病理变化，主要表现为核固缩或肿胀并出现核空泡，内质网膨胀、大部分转化泡状，线粒体溶解或固缩，溶酶体数量增多等。     

栉孔扇贝急性病毒性坏死症(AVND)病理学观察与分析

2000~2002年,应用光镜和电子显微技术对AVND发生期间栉孔扇贝(Chlamys farreri)各主要组织器官病理变化进行了连续观察。光镜观察显示,除生殖腺、闭壳肌外,濒死栉孔扇贝的外套膜、鳃、肾、消化腺和肠组织都有不同程度的组织病理变化。病灶主要出现在上述各器官的结缔组织以及上皮组织,病理变化表现为细胞核肿大、固缩、破裂,核染色质边缘化或空泡化,受感染细胞崩解、脱落,留下大片均质无结构的空白区域,形成凝固性坏死,结缔组织细胞质中有嗜碱性包涵体样颗粒存在。电镜观察显示,在病灶出现的组织细胞内,细胞核染色质异常凝集和边缘化、核膜周隙扩张或溶解。细胞器病理变化明显,内质网扩张,核糖体脱落,有髓鞘样小体出现;线粒体肿大、嵴融解,整个细胞出现解体现象。病变的结缔组织细胞与间质细胞细胞质中有大量直径为130~170nm、具有囊膜的球形病毒粒子存在,负染电镜观察表明,病毒囊膜表面覆有长20nm放射状纤突。病毒的发生基质在细胞质内,并被一膜性结构所包围。病毒在宿主细胞中的繁殖分为3个阶段,即病毒发生期、病毒装配期和病毒释放期。病理学观察证明,病毒感染与病理变化具有明显的相关性。     

大连地区中国对虾暴发性流行病病理学研究

本文报道１９９４年－１９９５年大连地区由杆状病毒引起的自然发病和人工感染实验中国对虾组织病理学研究。结果表明，自然发病虾和人工感染的虾组织病理相同，在胃、皮下、类淋巴、造血组织、鳃、中肠、后肠、肝胰腺、心脏等组织器官的上皮细胞、干细胞和结缔组织细胞中都可观察到显著胀大，呈空泡状或被苏木精浓染的细胞核。胃是病理变化出现最早且最严重的器官。在电镜下，胃、甲壳下上皮细胞核中可见大量长２７０～３５０ｎｍ，宽１００～１２０ｎｍ的杆状病毒颗粒，细胞核、线粒体出现胀大，嵴断裂、消失情况。干细胞、上皮细胞和疏松结缔组织细胞为靶细胞。     

纯化病毒复制中华绒螯蟹“颤抖病”的组织病理观察

采用石蜡切片及超薄切片技术，对人工感染纯化病毒后具典型颤抖症状的中华绒螯蟹进行病理学观察。结果显示：人工感染“颤抖病”病毒发病与自然发病的病理变化相似，病蟹的肝胰脏、鳃、心脏及腹神经节等组织器官发生了病变，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；超薄切片电镜下可见细胞内存在病毒样颗粒，线粒体内嵴受损严重。病变严重的区域，组织细胞坏死，结构崩解。提示肝胰脏、鳃、心脏及神经组织是病毒感染主要靶器官。     

文蛤牛首科吸虫寄生病的组织病理学

报道了一种牛首科（Bucephalidae)吸虫幼虫在文蛤体内的寄生情况以及所引起的组织病理学变化。观察结果表明,该吸虫幼虫主要侵占生殖腺,少部分进入附近的消化盲囊、鳃、外套膜和足等。消化道中未发现其存在。寄生虫不仅破坏已感染的组织,未感染部位的组织结构也呈现一定的病理学变化。严重感染时,生殖腺完全被吸虫幼虫侵占;消化盲囊、消化道和鳃等器官组织的上皮细胞脱落或溃散;肌肉紊乱或溶解等。严重者导致器官组织坏死,贝类死亡。     

载铜蒙脱石对嗜水气单胞菌粘附尼罗罗非鱼上皮细胞的影响

采用尼罗罗非鱼上皮细胞培养模型，观察嗜水气单胞菌对鳃上皮、皮肤上皮、肠上皮细胞的粘附率，研究载铜蒙脱石对细菌粘附的阻断作用及其对细菌粘附引起鱼上皮细胞膜生物特性变化的影响。结果显示：嗜水气单胞菌与鱼上皮细胞均有不同程度的粘附作用，粘附率大小顺序为鳃上皮〉皮肤上皮〉肠上皮细胞。载铜蒙脱石均显著降低了嗜水气单胞菌对鳃、皮肤和肠上皮细胞的粘附率（P〈0．05），而且对不同上皮细胞的粘附阻断率无显著差异。嗜水气单胞菌粘附肠上皮细胞后细胞膜生物特性发生了显著变化，与正常对照组相比，嗜水气单胞菌粘附鱼上皮细胞后，胞浆游离钙和细胞膜磷脂酶A2活性显著上升。载铜蒙脱石可显著降低由于嗜水气单胞菌粘附鱼上皮细胞引起的胞浆游离钙和细胞膜磷脂酶A2活性的升高。结果表明，载铜蒙脱石可有效阻断病原菌粘附，从而防治细菌感染和细菌移位。     

文蛤吸虫寄生病的组织病理学观察

报道了吸虫在文蛤(Meretrix meretrix Linnaeus)体内的寄生情况以及所引起的组织病理学变化。观察表明，该吸虫是以胚细胞方式繁殖，幼虫经历胞蚴阶段，发育成尾蚴。其寄生情况与寄生于其他双壳贝类的一些吸虫相似，均是以生殖腺为主要侵占部位，少部分则进入附近的消化盲囊、鳃、外套膜、足等，消化道中未发现其存在。已被感染的消化盲囊、外套膜等器官组织出现上皮细胞脱落或崩解，肌纤维排列紊乱或溶解，黏液细胞增生等病理症状。观察发现，寄生虫不仅破坏已感染的组织，未感染部位的组织也呈现一定的病理学变化，最终导致宿主因器官衰竭、功能丧失而死亡。     

赤点石斑鱼病毒性神经坏死症的组织病理和电镜观察

用逆转录-聚合酶链式反应（RT-PCR）检测患病赤点石斑鱼苗，呈Beta诺达病毒阳性。光学显微镜下观察到病鱼的脑、视网膜、脊髓有空泡．在脑部，空泡主要分布在端脑、间脑和小脑。受感染的细胞明显收缩、致密变化和嗜碱性。包涵体常为圆形，大小不一。透射电镜下，在感染细胞的细胞质可观察到含有病毒粒子的致密体。病毒粒子呈等面体，无外膜，直径为25～28nm，随机分布在细胞质或在致密体内排列成品格状。致密体大小不一。偶尔观察到较大致密体的外膜已破裂，病毒粒子被释放到细胞质。     

感染小瓜虫草鱼幼鱼组织病理学变化

应用组织病理学方法,对感染小瓜虫草鱼幼鱼的脑、心脏、鳃、鳍、头肾、中肾、上颌、皮肤肌肉及内脏团等组织器官进行观察。观察结果表明,各组织器官均有明显的组织病理学变化,鳃丝、上颌、胸鳍及背鳍等组织器官可见小瓜虫寄生;脑组织淤血;心耳、心室及动脉球内发生淤血;鳃丝淤血并有炎细胞浸润,部分鳃小片结构消失,呼吸上皮细胞脱落;头肾淤血;中肾肾小管、连接小管的上皮细胞内出现脂肪样变及玻璃样变,肾小球肿大导致肾小囊囊腔消失,肾间质淤血;腹皮下致密层胶原纤维发生玻璃样变,静脉淤血;肠道黏膜层部分上皮细胞脱落,严重者上皮细胞溶解消失,黏膜下层可见炎细胞浸润;肝胰脏内肝血窦、静脉淤血,肝脏局灶性坏死;脾脏内静脉淤血。     

感染嗜水气单胞菌鱼类组织病理变化与多组学分析的研究进展

嗜水气单胞菌感染引起的病害使我国鱼类养殖遭受了严重经济损失，制约了水产养殖业的健康发展。嗜水气单胞菌可产生多种毒力因子，致病过程及影响因素复杂，感染鱼类种类广。鱼类的肠道、脾脏、肝脏和肾脏具有重要的免疫功能，在抵御病原菌入侵中发挥重要作用，其中肠道菌群直接影响着宿主的免疫和炎症反应。本文着重阐述了嗜水气单胞菌感染后鱼类免疫相关器官的组织病理学变化、转录组水平基因的差异表达和通路富集及肠道微生物组成与结构改变，综述了嗜水气单胞菌感染后鱼类病理学和组学水平变化，为研究鱼类免疫应答机制及肠道菌群免疫调节功能提供参考依据。     

九孔鲍精子发生的超微结构

利用透射电镜观察九孔鲍（Haliotis diversicolor aquatilis）精子的发生过程。结果表明,九孔鲍精子的发生历经精原细胞、初级精母细胞、次级精母细胞、精细胞和成熟精子5个阶段。其中精细胞的分化可分为5期,主要特点为：Ⅰ期,一些高电子致密颗粒参与顶体的形成;Ⅱ期,核形态由椭圆形到近圆形,再逐渐拉长为长圆柱状;Ⅲ期,核染色质以颗粒状形式凝集,最后向高电子密度均质状态转变;IV期,线粒体逐步融合、体积增大和嵴发达;Ⅴ期,中心体移动及鞭毛形成,胞质逐渐减少。成熟精子为原生型,由头部、中段和尾部组成。通过观察精子发生过程中细胞形态结构的变化,分析了其演变规律,同时与软体动物的其他种类作了比较研究。     

3种珍珠贝的外套膜小片外表皮细胞的超微结构

用透射电子显微镜对合浦珠母贝（Ｐｉｎｃｔａｄａｆｕｃａｔａ）、大珠母贝（Ｐｉｎｃｔａｄａｍａｘｉｍａ）和企鹅珍珠贝（Ｐｔｅｒｉａｐｅｎｇｕｉｎ）的外套膜小片外表皮超微结构进行比较观察,发现都是单层柱状表皮,由３类细胞构成,即柱状表皮细胞、粘液细胞和嗜酸性粒分泌细胞。柱状表皮细胞是贝壳和珍珠组份的主要合成和转运者,据其结构和功能的差异可细分为Ａ、Ｂ、Ｃ３型。粘液细胞和嗜酸性粒分泌细胞呈杯状或不规则,体积巨大,细胞质稀少,核的形态亦不固定,前者的胞体全由电子透明泡占据,后者含有大量嗜酸性的电子稠密颗粒。这２类细胞的主要功能是参与物质转运,它们在邻近表皮的结缔组织中也有分布,可作变形运动穿越基膜进入上皮层。     

Morphogenesis of koi herpesvirus observed by electron microscopy

Koi herpesvirus (KHV or cyprinid herpesvirus 3) was inoculated onto three fish cell lines derived from carp, Cyprinus carpio L., and the process of virion formation observed with electron microscopy. Essentially, similar features of virus particles were observed in all three cell lines. The nucleus of infected cells was characterized by margination of chromatin and often contained many capsids of about 110 nm in diameter with varying morphology. The morphological variation of the capsids was very similar to that of mammalian herpesviruses. Some capsids protruded from the inner nuclear membrane, and others, with envelopes, were located in the perinuclear space between the inner and outer nuclear membrane, suggesting budding of capsids at the inner nuclear membrane. Naked capsids and envelopment of capsids by budding into vesicles were also observed in the cytoplasm. Mature, enveloped virions 170-200 nm in diameter were seen in cytoplasmic vesicles or outside the cell. These observations suggest KHV virions mature through a complex morphological pathway including two distinct envelopments, which have been found only in herpesviruses. These observations support the inclusion of KHV in the family Herpesviridae.     

Quantitative PCR demonstrates a positive correlation between a Rickettsia-like organism and severity of strawberry disease lesions in rainbow trout, Oncorhynchus mykiss (Walbaum)

Strawberry disease (SD) is an inflammatory skin disorder in rainbow trout, Oncorhynchus mykiss (Walbaum). The aetiology of SD is unknown although the 16S rDNA sequence of a Rickettsia-like organism (RLO) has been associated with SD lesions using a nested PCR assay. In this study, we developed a Taqman quantitative PCR assay (qPCR) that targeted the RLO 16S rDNA sequence to examine the distribution of RLO relative to lesion status. We compared 18 lesion samples from 13 fish representing high or low lesion severity as judged by gross examination. QPCR results showed that there was a higher number of RLO sequences in high severity lesions (mean of 12,068 copies) compared with fewer copies of RLO sequence in low severity lesions (mean of 3287 copies, P = 0.012). Grossly normal skin samples (n = 13) from SD-affected fish were all negative by qPCR except two samples (121 and 139 copies). The qPCR assay described herein is a useful tool to investigate the role of RLO in SD in the absence of a culture system for RLO. Our results demonstrate a positive correlation between copy number and lesion severity consistent with the hypothesis that the RLO is the aetiologic agent of SD.     

Haemopoiesis in the head kidney of tilapia, <Emphasis Type="Italic">Oreochromis niloticus</Emphasis> (Teleostei: Cichlidae): a morphological (optical and ultrastructural) study

The present work focused on the histological and ultrastructural studies on haemopoiesis in the kidney of tilapia, Oreochromis niloticus. Haemopoietic tissue was found mainly in the head kidney and a small amount occurred in the mesonephros. The haemopoiesis of tilapia had the following series: erythropoiesis, granulopoiesis, thrombopoiesis, monopoiesis and lymphoplasmopoiesis. Erythropoiesis includes proerythroblasts, basophilic erythroblasts, polychromatic erythroblasts, acidophilic erythroblasts and young and mature erythrocytes. The proerythroblasts were the largest cells in the erythropoietic series. During the maturation process both the nuclear and cellular size decreased gradually due to the chromatin condensation and the progressive substitution of cytoplasmic matrix with a large amount of haemoglobin. Granulopoietic series consisted of cells with variable shape and size at different stages of maturity from myeloblasts to mature granulocytes. The promyelocytes were the largest cells in the series and were characterised by the appearance of primary (azoruphilic) granules. The maturation process involved the appearance of specific granules in the heterophilic, eosinophilic and basophilic series. It is important to mention that eosinophilic granulocytes were the dominant granulopoietic series in the haemopoietic tissue (Ht) of tilapia. Lymphopoietic series consisted of lymphoblasts, large lymphocytes, small lymphocytes and active and inactive plasma cells. Thrombopoietic series consisted of thromboblasts, prothromboblasts and thrombocytes. Thrombocytes of tilapia were nucleated and possessed a spindle shape. Melanomacrophage centres were dominant among the Ht of the head kidney. Also, monocytes were detected and shown to be large cells with an indented nucleus and cytoplasm containing numerous vesicles of different sizes and a few lysosomes.     

Systematic granulomas caused by a rickettsia-like organism in Nile tilapia, Oreochronuis niloticus (L.), from southern Taiwan

Abstract. A rickettsia-like organism was observed in diseased Nile tilapia, Oreochronuis niloticus (L.), from southern Taiwan. Most affected fish were pale and lethargic with haemorrhages and ulcers on the skin. The most significant gross pathological changes were varying degrees of ascites, and enlargement of the spleen, kidney and liver. Marked white nodules (as ring-shaped foci), varying in size, were found in these organs. A Gram-negative, rickettsia-like organism (RLO) frequently appeared as inclusions or within host cell intra-cytoplasmic vacuoles. Fibrin thrombi, perivascular necrosis, chronic inflammatory cells with hypertrophy and RLO-laden cells were characteristic of the disease. White nodules induced in experimental fish were similar to naturally infected cases and RLOs were reisolated in tissue culture using a CHSE-214 cell line. This provides evidence that the systemic granulomas in Nile tilapia were caused by RLOs.     

皱纹盘鲍眼部组织的显微及亚显微结构观察

采用组织学和电镜的方法,对皱纹盘鲍(Haliotis discus hannai Ino)的眼部组织进行了光镜和电镜观察,以期为进一步从分子生物学角度解析鲍对光照的生理响应机制提供组织学与细胞学基础。结果发现,眼部组织由外至内依次为视网膜色素上皮细胞层、外核层、光感受器内节、内核层、黑色素颗粒沉积层、视觉纤维层。组织表面布满乳头状突起,每一乳状突起的顶端均具有一簇或两簇纤毛环。疏松结缔组织和平滑肌纤维等是组织内的主要成分,结缔组织间分布的胶原纤维等对保持细胞的弹性和韧性具有重要的作用。研究结果显示了鲍的眼部组织在感受和辨识外界光环境因子中的重要作用,也为鲍养殖生产中的光环境因子优化和调控、深入探讨鲍对光照的生理应答机制提供了形态学依据。     

坛紫菜自由丝状体细胞超微结构的初步研究

本文介绍了用扫描电镜和透射电镜对坛紫菜自由丝状体细胞观察的结果。发现在坛紫菜自由丝状体的膨大藻丝细胞外壁上具有脊突；在丝状藻丝及膨大藻丝细胞的间壁上，存在着孔状联系。在坛紫菜自由丝状体的细胞质中，清晰地观察到内质网结构；这些内质网多分布于细胞核的周围，有些地方并与核膜连接，在细胞膜的附近也能观察到内质网的存在。在细胞核中可以看到双层核膜，在双层核膜之间是一核膜间区，核膜上具有核孔，此外还可见到核仁和浓聚深色的染色质。     

Histopathological survey of diseases and pathogens present in redclaw crayfish, Cherax quadricarinatus (Von Martens), cultured in Ecuador

A histopathological survey of commercial farms in Ecuador culturing redclaw crayfish, Cherax quadricarinatus (Von Martens), revealed the presence of different viral, bacterial and non‐infectious pathologies. A total of 536 animals from 27 different farms were analysed during a 3‐year period; 25, 154 and 357 specimens in 1995, 1996 and 1997, respectively. From the total sampled, a virus referred to as Cherax quadricarinatus bacilliform virus (CqBV) had a prevalence of 44% in animals examined during 1996 and 33% in 1997. A rickettsia‐like organism (RLO) was detected in five specimens sampled from three different farms in 1995. This RLO was detected in 36 and 12% of samples analysed in 1996 and 1997, respectively, and was the infectious agent most frequently associated with mortalities above 60%. Co‐infection by CqBV and the RLO was observed in 18 and 3% of the crayfish examined in 1997 and 1998, respectively. Five specimens out of 25 examined presented no sign of an infectious agent during 1995. The proportion of specimens testing negative by histopathology to known or suspected infectious agents was 25 and 32% in 1996 and 1997, respectively. Although different infectious agents were present in cultured crayfish, farms in general had overall survivals higher than 50% in most ponds. Other conditions observed included an intracellular prokaryotic organism, iron granules in the R and F cells of the hepatopancreas, different species of epibionts on the surface of the gills and body, and two cases of haemocytic enteritis. Two cases of serious mortality in different farms were associated with the presence of Cowdry type A inclusions, pyknosis, karyorrhexis and haemocytic infiltration in the stomach hypodermis and the antennal gland.