犬传染性肝炎病毒的形态发生学及抗原定位研究

应用免疫金电子显微镜技术研究了犬传染性肝炎病毒(ICHV)在犬肾传代细胞内的增殖过程及其病毒抗原在感染细胞内外的定位.结果显示:(1)吸附在细胞膜表面的病毒粒子,主要通过细胞吞饮作用侵入细胞,而吸附在微绒毛膜表面的病毒粒子则可直接“渗入”细胞;(2)在病毒感染不同时用的细胞核内观察到均质型、微粒型、颗粒型、副晶格型、病毒包涵体型及致密型等6种类型包涵体,其中前5种能被免疫金标记,它们是尚未形成病毒粒子的抗原蛋白、病毒装配后剩余的抗原蛋白或ICHV的病毒粒子结晶,致密型包涵体未被免疫金标记,可能是病毒DNA;(3)感染细胞内出现的某些特殊结构均不具有病毒抗原性,其中纤维样结构和管状结构在病毒增殖过程中起支持作用.     

动物3种DNA病毒在宿主细胞内形态发生比较

对猪细小病毒(PPV)、牛疱疹病毒2型(BHV2)和犬腺病毒1型(CAV1)3种动物DNA病毒在宿主细胞内的增殖、释放方式以及所致细胞结构的变化,通过电镜进行了观察比较.(1)这3种动物DNA病毒的复制和装配过程均发生在细胞核内,以毒浆结构(Viroplast)或核内包涵体为增殖场所和物质基础,但并非都形成结晶样结构.(2)有囊膜的BHV2,其核壳体在细胞核内装配完成后,从核内膜上以出芽方式获得囊膜,然后进入核周池,聚集的病毒使核外膜向胞质方向隆起,形成病毒性包涵体而脱离核外膜,并逐渐向细胞膜的方向移动,最后从细胞膜的破损处以病毒包涵体形式释放到细胞间隙.而无囊膜的CAV1,核壳体在细胞核内装配完成后,从细胞核膜破损处或细胞核崩解后进入细胞质,待整个细胞崩解后才能释放出来.无囊膜的PPV,在核壳体装配完成后,成堆地以病毒流的方式,从扩张的核孔释放到细胞质中,待细胞崩解后再释放出来.(3)3种病毒增殖时,宿主细胞的固有细胞器,如线粒体、内质网以及溶酶体等均出现不同程度的超微结构变化,并能诱导宿主细胞出现一些新的结构,除毒浆结构外,还有管状结构、细纤维样结构、周期性结构和髓膜样结构等,其中周期性结构仅见于BHV2感染.     

MDCK细胞感染犬传染性肝炎病毒后的超微结构观察

MDCK细胞感染犬传染性肝炎病毒(ICHV)BJ—1株后6h未见明显改变;12～18h胞核肿大,核内形成黑色颗粒样球状体;24h胞核内偶见病毒粒子,36h许多细胞内含病毒、病毒副晶体、形态各异的包涵体和多种板层样结构;48h感染细胞核膜以破损或出芽方式将成熟病毒粒子释入胞浆.     

应用免疫胶体金标记技术对狂犬病病毒形态发生学的抗原定位研究

利用包埋后免疫胶体金标记技术，通过透射电子显微镜，对感染ＢＨＫ２１细胞的鹿狂犬病病毒进行了形态发生的抗原定位。结果：感染细胞的细胞质内有５种包涵体，其中４种被免疫胶体金特异性标记，这些病毒抗原是多成分的，另一种未被免疫胶体金标记的包涵体可能是病毒核酸；感染细胞的细胞质内大量堆积的病毒前体物质包涵体，其中可能有病毒装配后多余的成分，表明病毒各组分并不是按比例需要而合成的。     

伪狂犬病病毒的形态发生研究

病毒为圆形或卵圆形,直径为150～170nm,核壳体直径为94～103nm,内部充满致密的核心,在宿主细胞核内,可以观察到特异的毒浆结构和处于不同装配阶段的核壳体,有时在核膜内侧还可以见到被有囊膜的成熟病毒粒子。在细胞质中常常也可以见到装配好的核壳体和成熟病毒,但后者仅存在于细胞质的空泡中,成熟的病毒可能是通过与吞噬作用相反的过程逐渐向细胞外释放。膜增生现象和大量的纤维结构在病变的细胞质中是十分明显的。     

实验性犬传染性肝炎肝胆的扫描电镜和透射电镜观察

透射电镜下,犬传染性肝炎急性坏死型病例肝细胞线粒体高度肿胀,嵴断裂。病变严重的,肝细胞胞浆固有结构崩解、碎裂。核内包涵体由基质和ICHV粒子构成;在不含包涵体的核内也见到ICHV粒子。核内的ICHV粒子通过溶解的核膜或由于核崩解而释入胞浆;当细胞膜局部破裂或整个细胞崩解时,细胞内的ICHV粒子释放于细胞外。在嗜酸性小体的胞浆内观察到ICHV粒子。窦状隙内皮细胞和星状细胞呈现与肝细胞类似的变化。扫描电镜下,组织学呈凝固性坏死的胆囊粘膜袁面平坦无结构;亦见胆囊粘膜有局部灶坏死,与周围界限明显,坏死灶外围的上皮细胞结结构不整,上皮顶部凸出的颗粒大小不一或脱落。透射电镜下,呈坏死的胆囊粘膜上皮细胞固有结构消失,呈均质状;其余部位的上皮细胞微绒毛断裂、脱落;线粒体肿胀,嵴溶解,上皮顶部堆集大量的致密小体,细胞间连接复合体结构消失;亦见血管均质坏死,血管内皮的坏死与粘膜上皮的坏死同步或先于其发生。     

应用电镜原位杂交技术对宿主细胞内犬传染性肝炎病毒DNA定位

应用电镜原位杂交技术，在超微水平上对犬传染性肝炎病毒（ＩＣＨＶ）ＤＮＡ在感染的犬肾传代细胞内的分布进行了定位研究。结果胶体金颗粒不仅特异地标记在细胞核致密型包涵体上，而且在细胞质致密型包涵体上也有胶体金颗粒的特异标记，提示细胞质内含有大量ＩＣＨＶＤＮＡ成分，从而为ＩＣＨＶ的细胞质内装配途径提供了证据。     

犬传染性肝炎的诊疗及营养解决方案

<正>犬传染性肝炎是由犬传染性肝炎病毒(ICHV)引起的急性败血性传染病。ICHV又称为Ⅰ型犬腺病毒(CAV-Ⅰ)主要引起犬的急性肝炎,以肝小叶中心坏死、肝实质细胞和皮质细胞核内出现包涵体及出血时间延长为特征。临床诊断以血液生化指标和B超探查结果为依据,确诊需要     

犬传染性肝炎的诊治

犬传染性肝炎是由Rubarth于1947年首次发现的由犬传染性肝炎病毒（ICHV）引起的犬的一种急性、高度接触性败血性的传染病。该病的特征是循环障碍、肝小叶中心坏死、肝实质细胞和内皮细胞的核内出现包涵体。     

犬传染性肝炎的诊疗体会

<正>犬传染性肝炎是由腺病毒引起的一种急性败血性传染病,以肝小叶中心坏死,肝实质细胞和皮肤细胞内出现核内包涵体及出血时间延长为特征。犬传染性肝炎病毒(ICHV)属于腺病毒,犬腺病毒分为CA-Ⅰ型和CA-Ⅱ型。CA-Ⅰ型是引起犬传染性肝炎的病原,CA-Ⅱ型是引起呼吸道疾病的病原,因此,一般也将腺病毒引起的疾病分为肝炎型和呼吸型。该病毒抵抗力相当强,低温条件     

鸭病毒性肠炎病毒强毒在人工感染鸭体内形态结构和发生学的电镜观察

为研究鸭病毒性肠炎病毒(DEV)CH强毒株在感染鸭体内的分布和形态学发生规律，应用透射电镜和超薄切片技术对人工感染DEV的成年鸭各组织器官进行观察。结果表明：感染后12h在脾脏和法氏囊首先观察到少量的DEV出现，24h后在脾、胸腺和法氏囊以及死亡鸭的肝、肠和胰中均观察到具有典型的疱疹病毒粒子及其核衣壳形态的DEV。DEV病毒核衣壳有空心型、致密核心型、双环型和内壁附有颗粒型4种形态，存在胞核和胞浆两种装配方式。病毒成熟有两种方式：一为细胞核内核衣壳在核内获得皮层，通过核内膜获得囊膜成为成熟病毒；二为核内核衣壳通过内外核膜进入胞浆，核内和胞浆内的核衣壳在细胞浆中获得皮层，然后在各种质膜上获得囊膜，最后成熟病毒通过细胞破裂或其他方式释放到细胞外。伴随着病毒的复制、装配和成熟，细胞中出现多种核内和胞浆包涵体、核内致密颗粒、核内微管和中空短管、胞浆电子致密小体等结构。     

Electron microscopic studies of the morphogenesis of duck enteritis virus

The morphogenesis of duck enteritis virus (DEV) and distribution in vivo were observed by electron microscopy after ducks were infected experimentally with DEV virulent strain. The investigation showed that a few typical herpesvirus virions and nucleocapsids were first observed in the spleen, thymus, and bursa of Fabricius (BF), and many nucleocapsids, mature viruses, and viral inclusion bodies could be found in the nucleus and cytoplasm of infected liver, small intestine, spleen, thymus, and BF when the ducks died. Nucleocapsids assembled both in nucleus and cytoplasm and could be divided into four different types according to their structures. Typical herpesvirus, light particles (L-particles), and virions without tegument could be observed at the same time. With the replication, assembly, and maturation of the viruses, intracytoplasmic and intranuclear inclusion bodies, electron-density particles, microtubules, hollow tubes, and coated electron-density bodies were observed in infected cells.     

鸭病毒性肠炎病毒CH强毒株感染鸭胚成纤维细胞的超微结构观察

通过超薄切片和透射电子显微镜技术对鸭病毒性肠炎病毒（DEV）CH强毒株在鸭胚成纤维细胞（DEF）中的形态结构进行了研究。结果发现，DEVCH强毒株病毒核酸呈圆形颗粒状，直径35～45nm，在胞核内常集中分布；病毒核衣壳呈圆形．直径90～100nm．在胞核和胞浆内都有分布；DEV核衣壳可根据所含核酸形态的差异分为空心核衣壳、内壁附有颗粒型核衣壳、同心圆形核衣壳和实心核衣壳；成熟的病毒粒子具有囊膜和皮层结构．呈圆形．直径150～300nm，存在于胞浆空泡内；DEV可在DEF中分别形成胞浆内和胞核内包涵体结构；伴随子代病毒在细胞内的出现，胞浆内迁出现豆英状、马蹄形、半圆形、圆形、同心圆形等与病毒发生有关的电子致密结构。     

Morphologic observations of new type gosling viral enteritis virus (NGVEV) virulent isolate in infected duck embryo fibroblasts

The morphogenesis of the new type gosling viral enteritis virus (NGVEV) and the characteristic ultrastructural changes in the duck embryo fibroblasts (DEFs) were investigated by ultrathin sectioning and transmission electron microscopy after monolayer DEFs were experimentally infected with a virulent NGVEV strain. The investigation demonstrated that typical NGVEV particles were round, with a diameter ranging from 75 nm to 90 nm and that they were present in both the nucleus and cytoplasm of the infected DEFs. The mature virions contained nucleocapsids and nucleic acids. The virion penetrated the DEF, replicated, and matured in the nucleus, and they were finally released into the extracellular space via budding and disruption of the cytoplasmic membrane. With the appearance of progeny NGVEV, certain virus-related structures that were densely electron stained, which were circular, U-shaped, or irregular in appearance, could be observed in the cytoplasm of the infected DEFs. In this research, we first detected three types of intracytoplasmic inclusion bodies during the NGVEV infection, which always contained a number of NGVEV particles. Furthermore, we detected that NGVEV could induce apoptosis in DEFs, which had not been reported previously. The morphologic changes of apoptosis included shrinking of the apoptotic cells, chromatin condensation and margination, appearance of vacuoles on the cytoplasmic membrane, and the formation of apoptotic bodies. The mitochondria were ultracondensed and aggregated into compact clusters during apoptosis.     

Experimental viral pneumonia in guinea pigs: an ultrastructural study

Homogenized lung tissue was used to experimentally reproduce lethal viral pneumonia in guinea pigs. The resultant lesions corresponded with those of the spontaneous disease. Pneumonia with necrotic bronchiolar epithelium accompanied by basophilic intranuclear inclusion bodies was the primary finding. With transmission electron microscopy, numerous viral particles were found--mainly within the nucleus of pulmonary epithelial cells but also within the cytoplasm and in extracellular space. The appearance of viral particles, in particular their paracrystalline and crystalline deposition within the nucleus, indicates that our experimentally induced pneumonia was caused by an adenovirus.     

Filamentous Intranuclear Inclusion Bodies in Psittacine Birds. A Structural and Ultrastructural Study

This paper concerns a disease affecting a group of African grey parrots, which involves intranuclear inclusion bodies composed of filamentous material. The disease was characterized by either sudden death or death within 2–3 days from onset of non-specific symptoms. At necropsy, gross lesions included enlarged liver, mild hepatic congestion and focal necrosis. Samples from five birds were fixed in 10% formol and routinely processed for light and electron microscopy. In four birds, numerous hepatocytes displayed an enlarged nucleus, with peripheral margination of chromatin; the nucleus was partially or wholly filled by a basophilic inclusion body. In the remaining bird, inclusion bodies were acidophilic and completely filled the nucleus; nuclear enlargement was less evident than in the other birds. At ultrastructural examination, and in both types of IIB, nuclei contained looped filaments but no evidence of viral structures. However, virion-like structures were observed in the cytoplasm of some hepatocytes.     

Filamentous intranuclear inclusion bodies in psittacine birds. A structural and ultrastructural study.

This paper concerns a disease affecting a group of African grey parrots, which involves intranuclear inclusion bodies composed of filamentous material. The disease was characterized by either sudden death or death within 2-3 days from onset of non-specific symptoms. At necropsy, gross lesions included enlarged liver, mild hepatic congestion and focal necrosis. Samples from five birds were fixed in 10% formol and routinely processed for light and electron microscopy. In four birds, numerous hepatocytes displayed an enlarged nucleus, with peripheral margination of chromatin; the nucleus was partially or wholly filled by a basophilic inclusion body. In the remaining bird, inclusion bodies were acidophilic and completely filled the nucleus; nuclear enlargement was less evident than in the other birds. At ultrastructural examination, and in both types of IIB, nuclei contained looped filaments but no evidence of viral structures. However, virion-like structures were observed in the cytoplasm of some hepatocytes.