Chandipura Virus Forms Cytoplasmic Inclusion Bodies through Phase Separation and Proviral Association of Cellular Protein Kinase R and Stress Granule Protein TIA-1.

Negative-strand RNA viruses form cytoplasmic inclusion bodies (IBs) representing virus replication foci through phase separation or biomolecular condensation of viral and cellular proteins, as a hallmark of their infection. Alternatively, mammalian cells form stalled mRNA containing antiviral stress granules (SGs), as a consequence of phosphorylation of eukaryotic initiation factor 2α (eIF2α) through condensation of several RNA-binding proteins including TIA-1. Whether and how Chandipura virus (CHPV), an emerging human pathogen causing influenza-like illness, coma and death, forms IBs and evades antiviral SGs remain unknown. By confocal imaging on CHPV-infected Vero-E6 cells, we found that CHPV infection does not induce formation of distinct canonical SGs. Instead, CHPV proteins condense and co-localize together with SG proteins to form heterogeneous IBs, which ensued independent of the activation of eIF2α and eIF2α kinase, protein kinase R (PKR). Interestingly, siRNA-mediated depletion of PKR or TIA-1 significantly decreased viral transcription and virion production. Moreover, CHPV infection also caused condensation and recruitment of PKR to IBs. Compared to SGs, IBs exhibited significant rapidity in disassembly dynamics. Altogether, our study demonstrating that CHPV replication co-optimizes with SG proteins and revealing an unprecedented proviral role of TIA-1/PKR may have implications in understanding the mechanisms regulating CHPV-IB formation and designing antiviral therapeutics. Importance: CHPV is an emerging tropical pathogen reported to cause acute influenza-like illness and encephalitis in children with a very high mortality rate of ~70%. Lack of vaccines and an effective therapy against CHPV makes it a potent pathogen for causing an epidemic in tropical parts of globe. Given these forewarnings, it is of paramount importance that CHPV biology must be understood comprehensively. Targeting of host factors offers several advantages over targeting the viral components due to the generally higher mutation rate in the viral genome. In this study, we aimed at understanding the role of SGs forming cellular RNA-binding proteins in CHPV replication. Our study helps understand participation of cellular factors in CHPV replication and could help develop effective therapeutics against the virus.

Coordination of oxysterol binding protein 1 and VAP-A/B modulates the generation of cholesterol and viral inclusion bodies to promote grass carp reovirus replication.

Similar to other RNA viruses, grass carp reovirus, the causative agent of the hemorrhagic disease, replicates in cytoplasmic viral inclusion bodies (VIBs), orchestrated by host proteins and lipids. The host pathways that facilitate the formation and function of GCRV VIBs are poorly understood. This work demonstrates that GCRV manipulates grass carp oxysterol binding protein 1 (named as gcOSBP1) and vesicle-associated membrane protein-associated protein A/B (named as gcVAP-A/B), 3 components of cholesterol transport pathway, to generate VIBs. By siRNA-mediated knockdown, we demonstrate that gcOSBP1 is an essential host factor for GCRV replication. We reveal that the nonstructural proteins NS80 and NS38 of GCRV interact with gcOSBP1, and that the gcOSBP1 is recruited by NS38 and NS80 for promoting the generation of VIBs. gcOSBP1 increases the expression of gcVAP-A/B and promotes the accumulation of intracellular cholesterol. gcOSBP1 also interacts with gcVAP-A/B for forming gcOSBP1-gcVAP-A/B complexes, which contribute to enhance the accumulation of intracellular cholesterol and gcOSBP1-mediated generation of VIBs. Inhibiting cholesterol accumulation by lovastatin can completely abolish the effects of gcOSBP1 and/or gcVAP-A/B in promoting GCRV infection, suggesting that cholesterol accumulation is vital for gcOSBP1- and/or gcVAP-A/B-mediated GCRV replication. Thus, our results, which highlight that gcOSBP1 functions in the replication of GCRV via its interaction with essential viral proteins for forming VIBs and with host gcVAP-A/B, provide key molecular targets for obtaining anti-hemorrhagic disease grass carp via gene editing technology.

Pathological and phylogenetic characterization of a rare fowl adenovirus (FAdV-8b) associated with inclusion body hepatitis in naturally infected Meleagris gallopavo.

Adenoviruses are a diverse group of viruses that can cause a variety of diseases in poultry, including respiratory and gastrointestinal infections. In turkeys (Meleagris gallopavo), adenoviruses commonly cause hemorrhagic enteritis and, rarely, inclusion body hepatitis. In this study, we investigated fowl adenoviruses (FAdVs) circulating in turkeys in Egypt. Following clinical examination of 500 birds, a portion of the hexon gene was amplified from four out of 50 samples from diseased birds (8%), and one amplicon that produced a strong band was selected for sequencing. Molecular and phylogenetic analysis revealed that the virus in that sample belonged to serotype FAdV-8b. Histopathological and immunohistochemical examinations of prepared tissue sections were performed to confirm the pathological findings. Diseased birds exhibited ruffled feathers, low body weight, a crouching posture, and diarrhea. Gross examination revealed petechial hemorrhage on the spleen, swollen pale liver, and congested intestine. Microscopic examination revealed the presence of eosinophilic and basophilic intranuclear inclusion bodies, nuclear pyknosis, and apoptotic bodies in the liver, congestion, hemorrhage, and fibrosis in the lungs, and desquamation of enterocytes. The presence of viral antigens in the liver, lungs, and intestine was confirmed by immunohistochemistry. To our knowledge, this is the first report of the characterization of an outbreak of inclusion body hepatitis in turkeys (hybrid converter breeds) due to FAdV-8b in Egypt. This finding raises an epidemiological alarm, necessitating further studies, including full-genome sequencing, to trace the virus's origin and genetic diversity.

Investigating Liquid-Liquid Phase Separation in Virus-Generated Inclusion Bodies Using Fluorescence Recovery After Photobleaching of Fluorescently Labeled Host Proteins.

Many negative-sense single-stranded RNA viruses within the order Mononegavirales harm humans. A common feature shared among cells infected by these viruses is the formation of subcellular membraneless structures called biomolecular condensates, also known as inclusion bodies (IBs), that form through a process called liquid-liquid phase separation (LLPS). Like many other membraneless organelles, viral IBs enrich a specific subset of viral and host proteins involved in the formation of viral particles. Elucidation of the properties and regulation of these IBs as they mature throughout the viral replication process are important for our understanding of viral replication, which may also lead to the development of alternative antiviral treatments. The protocol outlined in this chapter aims to characterize the intrinsic properties of LLPS within the measles virus (MeV, a member of Mononegavirales) IBs by using an imaging approach that fluorescently tags an IB-associated host protein. This method uses common laboratory techniques and is generalizable to any host factors as well as other viral systems.

Vaccine Associated Measles Complicated by Suspected Measles Inclusion Body Encephalitis in a Pediatric Leukemia Patient and Stem Cell Transplant Recipient: A Focus on Clinical Evolution and Management.

BACKGROUND: Immunocompromised individuals are at increased risk for severe disease and complications from viral infections, highlighting the importance of vaccination. However, in extremely rare situations, vaccine associated viral infections can be associated with disseminated disease and complications in immunocompromised hosts. CASE: Herein, we present a case of a 1-year-old child diagnosed with acute myeloid leukemia less than 2 weeks after receiving live viral vaccines who developed acute vaccine-strain measles virus disease, later complicated by central nervous system involvement following hematopoietic stem cell transplantation. A brain biopsy specimen was positive for vaccine-strain measles virus detected by reverse transcriptase polymerase chain reaction. MANAGEMENT AND OUTCOME: She was treated with intravenous ribavirin, inosine pranobex, intrathecal interferon-alpha and donor lymphocyte infusion following measles-mumps-rubella vaccine boost. Despite these measures, the patient suffered neurologic decline and dysautonomia, expiring after compassionate extubation. Management and ideal risk mitigation strategies are discussed within the context of existing literature for this rare complication.

Ebola virus sequesters IRF3 in viral inclusion bodies to evade host antiviral immunity.

Viral inclusion bodies (IBs) commonly form during the replication of Ebola virus (EBOV) in infected cells, but their role in viral immune evasion has rarely been explored. Here, we found that interferon regulatory factor 3 (IRF3), but not TANK-binding kinase 1 (TBK1) or IκB kinase epsilon (IKKε), was recruited and sequestered in viral IBs when the cells were infected by EBOV transcription- and replication-competent virus-like particles (trVLPs). Nucleoprotein/virion protein 35 (VP35)-induced IBs formation was critical for IRF3 recruitment and sequestration, probably through interaction with STING. Consequently, the association of TBK1 and IRF3, which plays a vital role in type I interferon (IFN-I) induction, was blocked by EBOV trVLPs infection. Additionally, IRF3 phosphorylation and nuclear translocation induced by Sendai virus or poly(I:C) stimulation were suppressed by EBOV trVLPs. Furthermore, downregulation of STING significantly attenuated VP35-induced IRF3 accumulation in IBs. Coexpression of the viral proteins by which IB-like structures formed was much more potent in antagonizing IFN-I than expression of the IFN-I antagonist VP35 alone. These results suggested a novel immune evasion mechanism by which EBOV evades host innate immunity.

Cattle rabies: the effect of clinical evolution, viral genetic lineage, and viral load on the severity of histological lesions / Raiva bovina: relação do curso clínico, da linhagem genética do vírus e carga viral com a intensidade de lesões histológicas

Our objective was the characterization and staging of histological lesions in different anatomical sites of the central nervous system (CNS) of rabid cattle. The severity of the lesions was compared with the clinical stages of the disease, the variants of viral isolates, and with the load of virus. Thirty-one spontaneously affected rabid cattle the state of Santa Catarina underwent clinical follow-up and were eventually necropsied. CNS tissues were sampled and submitted to direct fluorescent antibody technique (DFAT), immunohistochemistry (IHC), routine histopathology with hematoxylin and eosin stain (HE), reverse transcriptase polymerase chain reaction (RT-PCR), and polymerase chain reaction in quantitative reverse transcriptase in real time (qRT-PCR). Affected cattle were allotted in four groups according to their clinical stage when euthanized: G1, euthanized while standing; G2, euthanized when in sternal recumbence; G3, euthanized when in lateral recumbence; and G4, affected cattle with natural death. In order to evaluate the degree of severity of the lesions and the presence of Negri bodies (NBs), the brain was sectioned at 9 sites. Additionally, spinal cord and trigeminal ganglion sections were examined. The intensity of the lesions was graded as either absent, mild, moderate, or marked, and the presence or absence of the NBs was noted. Histological lesions were characterized by lymphocytic and monocytic meningoencephalitis with NBs in 28 cases. In all analyzed groups, intensities of histological lesions ranging from mild to severe were observed. Brain regions with the highest inflammatory lesion intensity were the medulla at the level of obex, followed by the colliculus and thalamus. NBs were observed in a higher percentage in the cerebellum, followed by medulla at the obex level, striatum complex, and frontal telencephalon. The duration of the clinical course of the disease did not influence the intensity of the inflammatory lesion, but it did influence the presence of NBs, with a higher percentage of these inclusions in cattle that died naturally than in euthanized cattle. All isolated rhabdovirus included in this study were genetically compatible with samples from hematophagous bats Desmodus rotundus. The evaluation by qRT-PCR did not demonstrate a correlation between lesion intensity and the amount of virus.(AU)

Nosso objetivo foi a caracterização e estadiamento de lesões histológicas em diferentes locais anatômicos do sistema nervoso central (SNC) de bovinos raivosos. A gravidade das lesões foi comparada com os estágios clínicos da doença, as variantes dos isolados virais e com a quantidade de vírus. Trinta e um bovinos do estado de Santa Catarina, afetados naturalmente por raiva, foram acompanhados clinicalmente e, ao final, necropsiados. Os tecidos do SNC foram amostrados e submetidos a imunofluorescência direta, imunohistoquímica, histopatologia de rotina, reação em cadeia da polimerase via transcriptase reversa (RT-PCR) e reação em cadeia da polimerase em transcriptase reversa quantitativa em tempo real (qRT-PCR). Os bovinos afetados foram distribuídos em quatro grupos, de acordo com sua fase clínica: G1, eutanasiados quando ainda se mantinham em pé; G2, eutanasiados quando em decúbito esternal; G3, eutanasiados quando em decúbito lateral; e G4, bovinos afetados com morte natural. Para avaliar o grau de gravidade das lesões e a presença de corpúsculos de Negri (CNs), o cérebro foi seccionado em 9 locais. Além disso, seções da medula espinhal e do gânglio trigêmeo foram examinadas. A intensidade das lesões foi graduada como ausente, leve, moderada ou acentuada, e a presença ou ausência dos CNs foi anotada. Lesões histológicas foram caracterizadas por meningoencefalite linfocítica e monocítica com CNs em 28 casos. Em todos os grupos analisados foram observadas intensidades de lesões histológicas variando de leve a grave. As regiões cerebrais com maior intensidade de lesão inflamatória foram o bulbo no nível do obex, seguido do colículo e tálamo. CNs foram mais prevalentes no cerebelo, seguido pelo bulbo ao nível do óbex, corpo estriado e telencéfalo frontal. A duração do curso clínico da raiva não influenciou a intensidade da lesão inflamatória, mas influenciou a presença de CNs, com maior porcentagem dessas inclusões em bovinos que morreram naturalmente do que em bovinos sacrificados. Todos os isolados rabdovírus obtidos neste estudo eram geneticamente compatíveis com amostras provenientes de morcegos hematófagos Desmodus rotundus.(AU)

Distribuição do vírus rábico no sistema nervoso central em ruminantes naturalmente infectados / Distribution of rabies vírus in the central nervous system in naturally infected ruminants

Com o objetivo de identificar a distribuição das lesões do vírus rábico no sistema nervoso central de casos espontâneos de raiva em ruminantes e comparar as técnicas de imunofluorescência direta (IFD), inoculação em camundongos (ICC) e presença de corpúsculos de Negri para o diagnóstico da doença foram analisados materiais proveniente de 48 casos de raiva, incluindo amostras de córtex frontal, temporal, parietal e occipital, hipocampo, tálamo, colículo rostral e caudal, cerebelo, ponte, medula oblonga, núcleo da base e porções da medula cervical, torácica e lombar. De 48 amostras analisadas, todas foram positivas na IFD e na ICC e em 30 (62,5 por cento) foram encontrados corpúsculo de Negri (CN). No entanto, houve diferenças importantes no resultados dos três testes nas diferentes regiões do SNC avaliadas. Nos cortes de córtex cerebral, em 38 bovinos, a presença de corpúsculos de inclusão foi baixa (11 por cento-37 por cento) assim como a positividade para IFD e ICC (60-80 por cento). Pelo contrário, todas as amostras de ponte, tálamo e medula testados foram positivas para IFD e ICC. Em outras regiões do tronco encefálico e também no cerebelo a positividade para ICC e IFD foi de 60 por cento a 96,7 por cento. No cerebelo foi encontrada a maior frequência (88,2 por cento) de corpúsculos de inclusão. Em oito ovinos as provas de ICC e IFD foram positivas em todos os cortes e foram observados corpúsculos de inclusão em três animais. Foram analisados somente dois casos de caprinos encontrando-se corpúsculos de inclusão em um e ambos foram positivos para IFD e ICC. Os resultados obtidos nesse trabalho sugerem que a conduta recomendada pelo Manual Técnico de Controle da Raiva dos Herbívoros (MTCRH) permite o diagnóstico de raiva associando o estudo histológico aos testes de IFD e ICC que incluem cerebelo, tálamo e tronco encefálico que apresentam alta positividade para as provas de IFD e ICC. No entanto, a melhor conduta é a de incluir metade do encéfalo cortado longitudinalmente e amostras de medula. Isto permite examinar por IFD e ICC uma ou mais regiões onde essas provas apresentam maior positividade e, posteriormente, se essas provas fossem negativas, retornar ao material original e examinar outras regiões. Por outro lado, a coleta de amostras dos locais recomendados pela MTCRH, assim com a coleta de metade do encéfalo, podem prejudicar o diagnóstico de outras doenças para o qual é necessário o estudo de todo o encéfalo após a fixação em formaldeído, para constatar a simetria e a distribuição das lesões. Nestes casos, com base nos resultados obtidos neste trabalho, pode ser recomendado para diagnóstico laboratorial de raiva o envio exclusivo de porções da medula cervical, dorsal e lombar, já que as três porções apresentaram 100 por cento de positividade nas provas de IFD e ICC. Além disso, o estudo histológico de todas as porções do cérebro incluídas neste trabalho permitirá complementar o diagnóstico.

With the aim to study the distribution of lesions the rabies virus in spontaneous cases of rabies in ruminants and to determine the efficiency of the direct fluorescent antibody test (DFA), mouse inoculation (MI) and presence of Negri bodies in the diagnosis of the disease, 48 cases of the rabies were examined. Samples of frontal, temporal, parietal and occipital cerebral cortex, hippocampus, thalamus, rostral and caudal colliculi, cerebellum, pons, medulla oblongata, basal nuclei and sections of the cervical, thoracic and lumbar spinal cord were examined. Of the 48 samples examined all were positive on DFA and MI, and in 30 (62.5 percent) Negri bodies were observed. However there were differences in the results of the three tests among different regions of the central nervous system. In the samples of the cerebral cortex in 38 cattle, the frequency of inclusion bodies was low (11-37 percent), and so was the positivity to DFA and MI (60-80 percent). In contrast, all samples of thalamus, pons and spinal cord were positive to DFA and MI. In other regions of the brain stem, positivity to these tests varied between 60 percent and 96.7 percent. On histologic examination, the major frequency of Negri bodies (88.2 percent) was observed in the cerebellum. In eight sheep the DFA and MI tests were positive in all sections of the CNS examined and Negri bodies were found in three sheep. Only two goats were examined; both were positive in DFA and MI tests and in one Negri bodies were found. These results suggest that the recommendations of the Brazilian Technical Manual for Rabies of Herbivores is adequate for rabies diagnosis, because their recommendations include the histologic study and the examination of cerebellum, and sections of the brainstem with high positivity to DFA and MI tests. However, a better recommendation is to send for DFA and MI half of the brain cut longitudinally and samples of the spinal cord, which will permit to examine one or two sections, and if those are negative to get back to the material and examine the rest of the sections. In contrast, to collect samples of the brain or half brain can be inappropriate for the diagnosis of other diseases of the CNS, for which the study of the whole fixed brain is necessary to recognize the symmetry or distribution of lesions. In these situations by the results obtained here, it can be recommended to send different sections of the spinal cord for DIF and MI tests and to fix the whole brain for gross and histologic examinations.

Hallazgos patológicos en cerdos afectados con el síndrome del desmedro multisistémico postdestete de la República Argentina / Pathological findings in pigs affected by the postweaning multisystemic wasting syndrome in Argentina

El síndrome del desmedro multisistémico postdestete (PMWS) fue descrito por primera vez en Canadá en el año 1991 y desde entonces un número creciente de casos han sido diagnosticados en todo el mundo. En la Argentina, el PMWS fue reportado por primera vez recientemente. Se estudiaron 48 cerdos de 5 a 12 semanas de edad con signos característicos de PMWS procedentes de 19 granjas. Si bien se desconoce la distribución real del virus en nuestro país se observó desde el año 2001 un número creciente de granjas con PMWS y distribuidas en las principales provincias productoras. La histopatología fue una herramienta diagnóstica importante en casos sospechosos de PMWS con la observación de diferentes grados de lesión. En los animales estudiados las infecciones secundarias pudieron ser importantes, ya sea por patógenos oportunistas o por complicaciones bacterianas.

Postweaning multisystemic wasting syndrome was first described in Canada in 1991 and at present an increasing number of cases has been diagnosed worldwide. In Argentina the first cases of PMWS were reported recently. Forty eight 5 to 12 week old pigs with signs characteristic of PMWS from 19 farms were studied. Although the real distribution of the virus in our country is not known it was observed an increasing number of farms with PMWS distributed in the major producing provinces. The histopathology was an important tool in diagnosis of suspicious cases of PMWS with the observation of different degrees of lesion. In the studied animals, the secondary infections, either by opportunistic pathogens or secondary bacteria could be important.

Inclusion bodies induced by Bean rugose mosaic comovirus seen under light microscopy/

Two types of inclusion bodies were consistently observed under light microscopy in bean (Phaseolus vulgaris) leaf tissue infected with Bean rugose mosaic virus (BRMV), a species of the genus Comovirus, family Comoviridae. One type consisted of vacuolated inclusions found mainly in the cytoplasm of epidermal cells. The other type consisted of abundant crystalloid inclusions of different sizes and shapes found consistently in glandular hairs, guard cells, phloem tissue, xylem elements and occasionally in epidermal and mesophyll tissues. The two types of inclusion bodies stained with Azure A and Luxol Brilliant Green Bl-Calcomine Orange 2RS (O-G), and were similar to those seen to be caused by other species of comoviruses.

Un caso de encefalitis rábica de larga evolución: correlación clínico patológica / Human rabies encephalomyelitis of long survival: clinical pathological correlation

Se presenta el caso de un hombre de 22 años de edad que desarrolló encefalomielitis rábica de seis semanas de duración, y que fue tratado inicialmente como padecimiento psiquiátrico y después como encefalitis. Se destaca: 1.- El prolongado tiempo de evolución, ya que lo habitual es que la mayoría de los pacientes fallezcan durante los ocho días posteriores al establecimiento del diagnóstico. 2.- La evidente gliosis reactiva. 3.- Los cuerpos de inclusión en casi todos los cortes estudiados, tanto en la porción supra como en la infratentorial. 4.- Los cuepos de Negri únicos o múltiples y algunos gigantes en las neuronas del hipocampo, así como en las astas anteriores de la médula espinal. 5.- La proliferación de la microglia y la presencia de cuerpos acidófilos libres en el neurópilo. Todo lo anterior indica una larga evolución desde el punto de vista microscópico, lo que raramente se presenta en esta enfermedad. Se revisó la bibligrafía y se encontraron sólo diez casos de larga duración

Conceptos actuales sobre hepatitis viral / Current concepts on viral hepatitis

Contiene:Serologia de la hepatitis viral,metotos de diagnostico laboratorial en las hepatitis virales, nuevos metodos de diagnostico para el virus de la hepatitis B y C, estado actual de la hepatitis A, mutantes del virus B de la hepatitis y sus implicociones clinicas, infeccion por virus C, hepatitis E, manual de diagnostico y tratamiento de las hepatotias virales

Enfermedad por citomegalovirus (CMV): estudio morfológico de 24 casos de autopsia / Cytomegalovirus disease: pathological findings in 24 autopsies

Con el aumento de la sobrevida de pacientes inmunodeprimidos la enfermedad por CMV ha cobrado cada vez más importancia. Su diagnóstico se basa frecuentemente en estudios morfológicos. Los objetivos de esta revisión de autopsias fueron: a) delimitar los patrones morfológicos de la enfermedad por CMV; b) determinar el posible rol del CMV como causa de muerte y c) conocer la relación entre la condición de base del paciente y la extensión de la enfermedad. Se reunieron 24 casos. En 13 se encontraron inclusiones virales en más de un órgano. El pulmón fue el órgano más afectado (19 casos), seguido del hígado (7), suprarrenal y colon (5) y riñón, bazo e intestino delgado (4). En 20 casos (83 por ciento) se encontró una condición basal inmunodepresiva; neoplasia maligna (6 casos), sindrome de inmunodeficiencia (5), tratamiento inmunusupresor por patología no tumoral (4) y otros (5). El 54 por ciento de las autopsias presentó otras infecciones asociadas; micótica (8 casos), bacteriana (5), viral (2) y parasitaria (1). En 9 casos se consideró el CMV causa posible de muerte y en 5 fue la única causa posible encontrada. Los grupos con neoplasias y sindrome de inmunodeficiencia presentaron una enfermedad por CMV más extensa y el patrón pulmonar más frecuentemente encontrado como causa de muerte fue daño alveolar difuso y bronconeumonia extensa

Diagnóstico de hepatitis con cuerpos de inclusión mediante la prueba de inmunofluorescencia / Inclusion body hepatitis diagnosis by the immunofluorescence test

Se elaboró un conjugado para diagnóstico de hepatitis con cuerpos de inclusión (HCI). Para ello se utilizaron 60 pollos de engorda Arbor Acres, los cuales permanecieron en unidades de aislamiento durante 59 días. A partir de la 2a semana de edad, las aves fueron inmunizadas semanalmente con adenovirus tipo 1; se obtuvo un suero hiperinmune contra el virus utilizado. El suero obtenido permitió la elaboración de un conjugado contra adenovirus tipo 1. Se evaluó la especificidad del conjugado, usando como referencia 14 casos positivos a HCI diagnosticados por histopatología. Finalmente, se infectaron cultivos celulares de fibroblastos de embrión de pollo con adenovirus tipo I, luego se realizó en ellos la prueba de inmunofluorescencia, utilizando testigos negativos. Los resultados muestran que la técnica de inunofluorescencia es de gran confiabilidad para el diagnóstico de HCI.

Vacunas experimentales contra hepatitis a cuerpo de inclusión aviar. I valoración serológica / Experimental vaccines against avian inclusion body hepatitis. I. Serological evaluation

Se elaboraron dos vacunas experimentales (Vac-19 y Vac-28) de Hepatitis a Cuerpo de Inclusión (HCI) a partir de los serotipos 4,10 y 11, aislados en brotes de campo en el país. Las cepas se atenuaron por pasajes en huevos embrionados SPF (16 y 22 pasajes para ambas vacunas) y cultivo de células renales (3 y 6 pasajes respectivamente). Se utilizaron cuatro grupos (G) experimentales: G I y G II formado por 8 pollos cada uno, vacunados vía oral (4,4 DICC55/ml) con Vac-19 y Vac-28 respectivamente a la tercera semana y revacunados a los 35 días por igual vía y dosis. Los grupos G III y G IV con 5 pollos cada uno correspondieron a control positivo y negativo, ambos grupos sin vacunar, el primero solo desafiado con un pool de los serotipos 4,10 y 11, a los 45 días de edad y el último sin desafiar. Exámenes clínicos serológicos se realizaron previo y posterior a las vacunaciones y desafío. La respuesta inmune se midió, a través de Inmunodifusión en Gelosa (IDG) y Seroneutralización (SN) en Cultivo de Células Renales (CCR) por el método ß (200 DICC50/50*l). Mediante la prueba IDG se observaron reacciones positivas (100%) desde el día 43 en adelante en los grupos cavunados y desde el 55 en el grupo desafiado (GIII). Los Títulos Medios Geométricos (TMG) previo al desafío fueron G I de 1.194 y en G II 23.525. Con posterioridad al desafío se observó un TMG de 7.760 y . 305.736 respectivamente en ambos grupos. El G III tuvo un TMG de 640. No se observó signología clínica y mortalidad el desarrollo de la experiencia. Se concluye que solo Vac-28 originó protección a las aves, emdida por la respuesta inmune humoral, pero por las lesiones observadas en el grupo solo vacunado se debería continuar estudiando sucompleta atenuación

Diagnóstico citológico y ultraestructural de infección por virus polioma humano (BK) en el sedimento urinário / Cytological and ultrastructural diagnosis of human polyoma virus infection (BK) in the urinary sediment

En 1971 se aislaron dos virus Polioma humanos: JC causante de la leucoencefalopatía multifocal progresiva y BK que están asociados a estenosis ureteral en algunos pacientes trasplantados renales. Los enfermos que hacen reactivación de la infección excretan el virus por la orina y descaman células uroteliales con típicas inclusiones diagnósticas. En caso de duda sobre el virus responsable de estas inclusiones, puede recurrirse a la ME o a estudios serológicos. Otro diagnóstico diferencial es con el cáncer urotelial. La reactivación ocurre en inmunodeficientes y también en pacientes con afecciones clínicas sin obvia deficiencia inmunológica. La citología puede utilizarse para pesquisa catastral. Se presenta un caso de infección por virus Polioma humano (*BK) diagnosticado por la citología urinaria y confirmado por ME, con el objeto de hacer conocer esta patología en nuestro medio y su posible confusión con cáncer urotelial en el citodiagnóstico

Raiva humana. Estudo neuropatologico de trinta casos. / Human rabies. Neuropathological study of 30 cases

O estudo epidemiologico e neuropatologico concernente a trinta autopsias de encefalite rabica foi realizado, tendo sido os dados obtidos comparados com aqueles da literatura. Nao houve relacao estatistica entre a topografia lesional e o local da mordida, entre o periodo de incubacao ou a duracao da doenca e a presenca ou ausencia de inclusoes virais ((IV) assim como entre a intensidade do processo inflamatorio (PI) e a presenca de IV. A dispersao do PI e das IV atraves o SNC foi diretamente proporcional a sua intensidade. A estrutura mais frequentemente comprometida pelo PI foi o mesencefalo, seguido pelo bulbo, ponte e medula espinhal. A estrutura mais intensamente afetada foi o bulbo. As IV foram particularmente proeminentes no hipocampo e cerebelo