Probable herpesvirus infection in an eastern cottontail (Sylvilagus floridanus).

One wild eastern cottontail (Sylvilagus floridanus) from Milwaukee County, Wisconsin was necropsied. The lungs contained numerous multifocal, circumscribed, tan foci; the spleen was markedly enlarged and had a mottled reddish tan color; and the brain had a red to tan friable tract in the left hemisphere. Microscopically, the lung had a severe bronchiolitis and pneumonia. The bronchiolitis was characterized by epithelial cells containing eosinophilic intranuclear inclusion bodies. The encephalomalacia of the left cerebral cortex featured tissue disruption and astrocytes or neurons containing intranuclear inclusion bodies. Herpesvirus particles were found within the bronchiolar epithelial cells. Based on histopathological and ultrastructural findings, a herpesvirus seemed the most likely etiologic agent.     

斜带髭鲷外周血嗜中性粒细胞核内包涵体的超微结构

目的在对斜带髭鲷（Hapalogenys nitens）外周血细胞结构进行透射电镜观察时发现嗜中性粒细胞存在大量的核内包涵体,系统研究了这些核内包涵体的超微结构,以探讨其来源和形成过程。方法应用电镜技术对这些核内包涵体的超微结构进行研究。结果斜带髭鲷外周血嗜中性粒细胞的核内包涵体可分为假包涵体和真包涵体两种类型,包涵体中的内含物来自胞质。胞核首先是以核膜内陷的方式将胞质及其各种有形成分包绕进核内而在核质外层形成具有双层膜包裹的典型假包涵体,随后假包涵体双层膜降解消失而转化成无被膜包裹的真包涵体,即核内糖原包涵体。结论假包涵体是形成真包涵体的开始阶段。随着假包涵体向真包涵体的转变,包涵体内含物的组成及其超微结构也出现了显著变化。     

Intranuclear filamentous inclusions in the gastro-entero-pancreatic (GEP) endocrine cells of birds

Summary Intranuclear filamentous inclusions were found in the normal endocrine cells of the avian stomach and pancreas. These inclusions were composed of a bundle of closely packed filaments (6–8 nm in diameter), being ultrastructurally similar to those found in the nucleus of various neurons. Most of them appeared as single rod- or spindle-shaped bodies; aggregations of two or more inclusions were rarely seen within a single nucleus. Cells with an intranuclear inclusion often contained a cytoplasmic fibrillar bundle similar to the intranuclear inclusion.     

Electron Microscopy of Measles Virus Replication

Replication of measles virus in HeLa cells was examined by electron microscopy with ultrathin sectioning and phosphotungstic acid negative staining methods. The cytoplasmic inclusion bodies consisted of masses of helical nucleocapsid which was similar in structure to the nucleocapsid found in measles virions. The cytoplasmic helical nucleocapsid appeared to align near the HeLa cell membrane, and the membrane differentiated into the internal membrane of the viral envelope and the outer layer of the short projections. The viral particles were released by a budding process involving incorporation into the viral envelope of membrane which was contiguous to but morphologically altered from the membrane of the HeLa cells. The intranuclear inclusion bodies were composed of tubular structures similar to those found in the cytoplasmic inclusion bodies. These structures aggregated to crystalline arrangement. The relationship between nuclear inclusion body and replication of measles virus was not clear.     

Histopathology of feline panleukopenia in domestic cats.

Histopathological observations were carried out on 17 domestic cats naturally affected with feline panleukopenia. Principal lesions were found in the intestine, bone marrow, and lymphoid organs. Intestinal lesions were characterized by degenerative changes accompanied by the appearance of intranuclear inclusion bodies in the epithelial cells of the crypts. In contrast to the crypts, the villi were seldom involved. Hypoplasia, parenchymal degeneration, and activation of the reticuloendothelial system were observed in the bone marrow and lymphoid organs. Intranuclear inclusion bodies were found occasionally also in the reticular and parenchymal cells of the bone marrow, lymphoid organs, liver, adrenals, and pancreas. Most of the inclusion bodies were amphophilic when stained with hematoxylin and eosin and occupied the whole area of the nucleus without producing any zone of clear halo. While cells bearing inclusion bodies underwent degenerative changes constantly in the intestinal crypts, the formation of inclusion body was not accompanied by the degeneration of corresponding cells in any other organ. Pathological changes as mentioned above were considered to be closely related to the systemic infection of feline panleukopenia virus.     

A parvo-like virus disease of penaeid shrimp

Cultured populations of four penaeid shrimp species (Crustacea, Decapoda) from four separate culture facilities in Asia were found to be adversely affected by a disease of presumed viral etiology. Individual shrimp with the disease displayed nonspecific signs, including poor growth rate, anorexia, reduced preening activity, increased surface fouling, and occasional opacity of tail musculature. These signs were accompanied by mortalities during the juvenile stages, after apparently normal development through the larval and postlarval stages. Accumulative mortality rates in epizootics in Penaeus merguiensis and P. semisulcatus reached as high as 50 to 100%, respectively, of the affected populations within 4 to 8 weeks of disease onset. The principal lesion, common to all four species, was necrosis and atrophy of the hepatopancreas, accompanied by the presence of large prominent basophilic, PAS-negative, Fuelgen-positive intranuclear inclusion bodies in affected hepatopancreatic tubule epithelial cells (hepatopancreatocytes). These inclusion bodies presumably developed from small, eosinophilic, intranuclear bodies that were also present in the affected tissues. Electron microscopy of affected hepatopancreatocytes revealed aggregations of 22- to 24-nm-diameter virus particles within the electron-dense granular inclusion body ground substance. The virus particle size and morphology, the close association of the nucleolus with the developing inclusion body, and the presence of intranuclear bodies within developing inclusion bodies are similar to cytopathological features reported for parvovirus infections in insects and vertebrates. It is suggested that this presumed virus disease of cultured penaeid shrimp be called HPV for Hepatopancreatic Parvo-like Virus disease.     

Origins and formation of intracellular inclusions associated with two leguminous virus diseases

Summary Two hitherto undescribed viruses were isolated from naturally-infected white clover plants.One induced both cytoplasmic and intranuclear inclusions, and the other caused cytoplasmic inclusions of a new type, which we have called Corner inclusion bodies.All three kinds of inclusion were found in the same cells.Vital observations showed that the two kinds of cytoplasmic inclusion bodies developed principally from the plastids.The intranuclear inclusions were shown to develop from the nucleoli.Cytochemical tests showed that the cytoplasmic inclusions contained protein and ribonucleic acid.On the basis of these findings, the possibility of virus synthesis in plastids and nucleoli is discussed.     

Unidentified kidney inclusion bodies in a lead-poisoned lorikeet

A lorikeet (unidentified species of subfamily Loriinae), which died of coliform septicemia subsequent to an esophageal ulcer also had intranuclear inclusion The ultrastructure bodies in the proximal convoluted tubular lining cells of the kidney. The ultrastructure of these inclusion bodies was compared with those described in experimental lead-induced inclusion bodies of rats.     

Cell-Free Transmission and In Vivo Replication of Marek''s Disease Virus

Marek's disease virus recovered from the feather follicle of infected chickens was found to be infectious for chickens in cell-free preparations. The virus replicated in epithelial cells of the germinative layer of the feather follicle epidermis, producing both intranuclear and round or diffuse cytoplasmic inclusion bodies in the infected cells. It was found at this site 2 weeks postinoculation and prior to the development of tumor or other gross lesions. In the nucleus, many naked and a few enveloped herpesvirions were found, whereas the cytoplasm contained predominantly enveloped herpesvirions, which were usually within the cytoplasmic inclusion bodies. Approximately 80% of the extracellular virions were enveloped. Studies with both virulent and avirulent strains of the virus revealed a relationship between virulence, contagiousness, and replication of the virus in the feather follicle.     

Karyokinetic reactions in rat kidney proximal convoluted segment epithelium on long-term lead supply with drinking water

In the rat kidney proximal convoluted segment epithelium the development of intranuclear inclusion bodies was observed after long-term lead administration with the drinking water. The inclusion bodies were PAS-positive. By electron microscopy they were identified as composed of filaments and granules lacking any kind of surrounding membrane. During the experiment, the nuclear volumes increased commensurately with the duration of lead exposure. The nuclear volume increase was considered a physiological reaction since it began long before the first appearance of intranuclear inclusion bodies.     

Microscopic evidence of adenoviral infection in a muskrat in Illinois

A wild muskrat (Ondatra zibethicus) found moribund in Illinois (USA) had minimal meningitis and pleuritis, probably of bacterial origin. There were large, basophilic, intranuclear inclusion bodies within scattered enterocytes. The inclusions were microscopically typical of those produced by adenoviruses, and ultrastructurally were intranuclear paracrystalline arrays of virus particles with characteristics of adenoviruses. The significance of the adenovirus infection in this muskrat is unknown.     

Fatal herpesvirus tamarinus infection in cotton-topped marmosets (Saguinus oedipus).

Fatal herpesvirus tamarinus infection was observed in cotton-topped marmosets (Saguinus oedipus) imported from South America via the United States on August 26, 1976. In addition to the lesions hitherto reported in herpesvirus tamarinus infection, severe degenerative and necrotic changes of ganglion cells were recognized with intranuclear inclusion bodies in the plexus of the digestive tract and the sympathetic nerves and their ganglions in the abdominal cavity. Inflammatory or regressive changes were also noted in the central nervous system. A large number of basophilic or eosinophilic intranuclear inclusion bodies frequently recognized in multinucleated giant cells were observed in various organs and tissues, and they showed different shapes at the electron microscopic level. Morphological findings indicated that herpesvirus tamarinus infection seemed to be similar to herpes simplex virus infection in man. The findings of the susceptibility of a variety of cell cultures to the virus isolate serologically identified as herpesvirus tamarinus and physicochemical characteristics of the virus isolate were in general agreement with the findings of herpesvirus tamarinus already reported by previous workers.     

Chronic lead intoxication causes a brain-specific nuclear protein to accumulate in the nuclei of cells lining kidney tubules

A characteristic feature of chronic lead intoxication is the induction of intranuclear inclusion bodies in cells lining kidney proximal tubules. These are relatively insoluble lead- and protein-rich structures which may serve a protective function by sequestering lead. The most abundant protein of isolated inclusion bodies, p32/6.3, has been partially characterized by use of a monoclonal antibody. As predicted by biochemical analysis, p32/6.3 occurs in kidney only in conjunction with lead intoxication and inclusion body formation. It does not accumulate in other tissues as a result of lead exposure. Unexpectedly, p32/6.3 was found to be a constitutive protein of adult brain, occurring primarily in the cerebral cortex. Within this tissue, both neurons and astrocytes contained p32/6.3. The brain p32/6.3 was concentrated in the insoluble nuclear protein or matrix fraction, a localization reminiscent of the intranuclear inclusion bodies from lead-exposed kidney. Brain p32/6.3 was detected in rat, mouse, dog, man, and chicken. In rat brain, the appearance of p32/6.3 was developmentally regulated. Only traces were detected 3 days after birth but within 1-2 weeks adult levels were achieved. The presence in brain of a protein which is involved in a potentially protective response to lead suggests that the brain may also have such a protective mechanism.     

Lethal pneumonia in guinea pigs associated with a virus

No bacteria were observed in an epizootic of lethal pneumonia in guinea pigs. Necrotic bronchitis and bronchiolitis with basophilic intranuclear inclusion bodies in bronchial epithelial cells were characteristic. Although adenovirus infection of guinea pigs has not previously been reported, histological findings paralleled those found in adenovirus infections of other animals including man. Virus particles found by electron-microscopical examination of the lung tissue closely resembled adenoviruses. The disease seemed to have a low contagiousness, a low morbidity (about 0.7%), but an acute course and a high mortality (100%).     

Detection of hepatopancreatic parvovirus (HPV) of penaeid shrimp by in situ hybridization at the electron microscope level

A post-embedding in situ hybridization procedure was developed to detect hepatopancreatic parvovirus (HPV) of penaeid shrimp at the ultrastructural level. The procedure was optimized using sections of resin-embedded hepatopancreas from HPV-infected juvenile Penaeus monodon and postlarval P. chinensis. The hepatopancreata were fixed using various fixatives, dehydrated, and embedded in the hydrophilic resin Unicryl. A 592 bp HPV-specific DNA probe, labeled with DIG-11-dUTP, was tested both on semi-thin and ultra-thin sections and examined by light and electron microscopy, respectively. Hybridized probe was detected by means of an anti-DIG antibody conjugated to 10 nm gold particles and subsequent silver enhancement. Hybridization signal intensities were similar with all fixatives tested, but ultrastructure was best preserved with either 2 or 6% glutaraldehyde. Post-fixation with 1% osmium tetroxide improved ultrastructure but markedly decreased hybridization signal and induced non-specific deposition of gold and silver. Under optimized conditions, this technique was used to successfully follow the development of HPV from absorption and transport through the cytoplansm to nuclear penetration, replication and release by cytolysis. The probe signal was consistently observed among necrotic cell debris within the lumen of hepatopancreatic tubules, within the microvillous border of tubule epithelial cells, within the cytoplasm, and within diagnostic HPV intranuclear inclusion bodies. The nucleolus and karyoplasm of patently infected cells (i.e., showing HPV intranuclear inclusion bodies) were almost devoid of signal. Electron-lucent structures, known as intranuclear bodies, commonly found within the virogenic stroma, showed only weak labeling. This is the first use of in situ hybridization to detect HPV nucleic acids with the electron microscope. The technique should be useful for studying the pathogenesis of HPV.     

Herpesvirus-like infection in a painted turtle (Chrysemys picta)

A painted turtle (Chrysemys picta) which died in captivity had marked necrosis in the liver and lungs with numerous intranuclear inclusion bodies in hepatocytes and respiratory epithelial cells. Electron microscopy revealed herpesvirus-like particles in cells in affected tissues.     

Papovaviral sialoadenitis in athymic nude rats

A wasting disease was found in 32 athymic nude rats. The rats had parotid sialoadenitis with intranuclear inclusion bodies in ductal and acinar epithelial cells. Other common lesions included bronchitis, bronchiolitis and secondary bacterial pneumonia. Less commonly, rhinitis and Harderian adenitis were seen. Intranuclear inclusions were also seen in bronchial epithelium of 1 rat, Harderian gland acini of 1 rat and laryngeal glands of 2 rats. Viral particles, averaging 45 nm in diameter, sometimes in crystalline arrays, were found in the nucleus of parotid epithelial cells. By the use of the avidin-biotin-peroxidase complex (ABC) immunoperoxidase technique, antibodies to disrupted SV40 virus (the group specific antigen of the polyomavirus (miopapovavirus) genus of the papovavirus family) reacted with intranuclear inclusions and cytoplasm of parotid epithelium and inclusions in lung and Harderian gland. The viral antigen did not cross react with antibodies to mouse polyoma, mouse K or disrupted bovine papilloma viruses.     

Cytomegalic inclusion disease in the east African headgehog.

Cytomegaly with intranuclear inclusion bodies was found in the duct epithelium of the salivary glands of five of ten hedgehogs (Atelerix albiventris). Cytomegalovirus infection was presumed to be the cause.     

Intranuclear inclusions containing melanosomes observed after PUVA therapy: a study of five psoriatic patients

Skin biopsies were obtained before and after PUVA therapy from five psoriatic patients and epidermal melanocytes surveyed for the presence of intranuclear melanosome-like bodies and nuclear bodies. Intranuclear melanosome-like bodies were observed after PUVA therapy whereas none were found before therapy. Nuclear bodies were found to increase in frequency after PUVA therapy. Remnants of a nuclear envelope membrane were not found around the intranuclear melanosome-like bodies. Nor were remnants of a nuclear envelope membrane found in or around the nuclear bodies. The results are discussed in view of the possible sites of origin of intranuclear melanosome-like bodies and nuclear bodies.     

Histopathology of hemorrhagic enteritis in turkeys.

In the summer of 1972, a disease of turkeys manifesting hemorrhagic diarrhea as a main clinical symptom and hemorrhagic enteritis as a necropsy finding broke out on a turkey farm in Japan. Seven-week-old Large White turkeys suffering from this disease were studied histopathologically and electron microscopically. Clinically, affected birds showed bloody diarrhea. Death occurred to them after an acute course. In the blood film, immature monocytes were higher in count in them than in healthy birds. Necropsy revealed a number of dark red bloody clots in the intestinal tract, many petechiae in the mucous membrane of small intestine and ceca, and atrophy of the spleen. The histopathological changes characteristic of this disease were acute hemorrhagic enteritis, degenerative changes of lymphatic tissue, proliferation of reticuloendothelial cells all over the body, and formation of intranuclear inclusion bodies in these cells. By electron microscopy, viral particles showing a crystalline array were found in the nuclei of reticuloendothelial cells. Viral particles which had electron-dense nucleoids and a naked hexagonal shape were about 80 nm in average diameter. The ultrastructural features of those inclusion bodies were identical with those of avian adenovirus.