RESUMEN
Different rhabdoviruses have been found in healthy bats, suggesting asymptomatic infection. The aim of this study was to focus on the epidemiology and pathogenesis of EBLV1 infection in the meridional serotine bat (Eptesicus isabellinus), as well as to search for other rhabdoviruses in this bat, which is the responsible for more than 95% of cases of human exposure to lyssaviruses in Europe. RT-PCR on oropharyngeal swabs was used together with antibody detection by the Rapid Fluorescent Focus Inhibition Test (RFFIT) to investigate EBLV1 circulation in 19 natural colonies of meridional serotine bats in Andalusia (Spain) from 1998 to 2003. The survey was based on 1,227 different captures of 1,033 individuals that were ring banded, sampled and released. Individuals that were repeatedly captured were always found in the same colony, despite the fact that some colonies were less than five km apart. Viral circulation was detected in ten colonies either by RT-PCR, serology or both. Each colony showed a different temporal pattern of viral circulation suggesting independent endemic circulation. Some positive individuals were captured healthy in following campaigns providing evidence for survival after viral infection. RNA from two apparently new Dimarhabdoviruses was also found in the pharyngeal cavity of two healthy bats.
Asunto(s)
Quirópteros/virología , Lyssavirus/aislamiento & purificación , Infecciones por Rhabdoviridae/veterinaria , Animales , Animales Salvajes , Lyssavirus/clasificación , Lyssavirus/inmunología , Orofaringe/virología , ARN Viral/química , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Infecciones por Rhabdoviridae/diagnóstico , Infecciones por Rhabdoviridae/epidemiología , Infecciones por Rhabdoviridae/virología , España/epidemiologíaRESUMEN
The aim of this study was to report on the lesions occurring in the central nervous system (CNS) during experimental classical swine fever (CSF) to clarify the spatial and chronologic distribution of the lesions and virus antigen in the CNS. To learn more about the pathogenetic mechanisms of the lesions during CSF in the CNS and to investigate the role of the virus in these mechanisms, cellular infiltrates and infected cells have been characterized. Twenty-eight pigs were inoculated with the virulent CSF virus isolate Alfort 187 and slaughtered from 2 to 15 postinoculation days; 4 animals of similar background served as a control group. Immunohistochemistry, electron microscopy, and the transferase-mediated deoxyuridine triphosphate nick-end labeling method were used to detect viral antigens and apoptosis. The results showed the presence of nonpurulent meningoencephalitis, occasional microhemorrhages, and apoptosis of the lymphocytes forming the perivascular and interstitital infiltrate in swine with CSF. Macrophages appeared to display little involvement in CNS lesions. The infected cells observed at the early stage of disease were lymphocytes and microglial cells in the rostral portion of the telencephalon, with infection of these cells in other areas in the next stages. The relationship between these lesions and the presence of viral antigen varied according to the type of lesion: hemorrhages were not associated with the presence of antigen in endothelial cells, but infiltrate-cell apoptosis was temporally and spacially associated to viral infection. However, the link between viral infection and the presence of cell infiltrate was far from clear.
Asunto(s)
Enfermedades del Sistema Nervioso Central/veterinaria , Sistema Nervioso Central/patología , Virus de la Fiebre Porcina Clásica/crecimiento & desarrollo , Peste Porcina Clásica/patología , Animales , Antígenos Virales/análisis , Apoptosis/fisiología , Encéfalo/patología , Encéfalo/ultraestructura , Encéfalo/virología , Sistema Nervioso Central/virología , Enfermedades del Sistema Nervioso Central/patología , Enfermedades del Sistema Nervioso Central/virología , Peste Porcina Clásica/virología , Femenino , Histocitoquímica/veterinaria , Etiquetado Corte-Fin in Situ , Masculino , Microscopía Electrónica de Transmisión/veterinaria , PorcinosRESUMEN
The aim of this study was to report on the lesions occurring in the intestine during experimental classical swine fever (CSF) and to clarify the nature of infected cells and the distribution of viral antigen. Thirty-two pigs were inoculated with the virulent CSF virus (CSFV) isolate Alfort 187 and slaughtered from 2 to 15 postinoculation days; four animals of similar background served as a control group. Immunohistochemistry, electron microscopy, and the transferase-mediated deoxyuridine triphosphate nick-end labeling method were used to detect viral antigens and apoptosis. The results showed progressive lymphoid depletion and mucosal necrosis. The lymphoid depletion could have been caused by apoptosis of lymphocytes but could not be directly attributed to the effect of CSFV on these cells. Vascular changes, pathogenic bacteria, and viral infection of epithelial cells were ruled out as causes of necrotic lesions. However, large virally infected monocytes-macrophages with ultrastructural changes indicative of activation were observed in the intestine. This suggests that monocytes-macrophages play an important role in the pathogenesis of intestinal lesions. An understanding of the function of these cells will require additional study.
Asunto(s)
Virus de la Fiebre Porcina Clásica , Peste Porcina Clásica/patología , Enfermedades Intestinales/veterinaria , Mucosa Intestinal/virología , Tejido Linfoide/virología , Animales , Antígenos Virales/metabolismo , Apoptosis/fisiología , Temperatura Corporal , Peso Corporal , Peste Porcina Clásica/virología , Femenino , Inmunohistoquímica/veterinaria , Etiquetado Corte-Fin in Situ/veterinaria , Enfermedades Intestinales/patología , Enfermedades Intestinales/virología , Mucosa Intestinal/ultraestructura , Tejido Linfoide/ultraestructura , Masculino , Microscopía Electrónica/veterinaria , Ganglios Linfáticos Agregados/ultraestructura , Ganglios Linfáticos Agregados/virología , PorcinosRESUMEN
This paper describes major pathogenetic mechanisms of African and Classical Swine Fever virus infections. The interactions between both viruses and the monocyte-macrophage-system result in the release of mediator molecules, which are important for the further progression of the diseases. The causes of the thrombocytopenia and the mechanisms of the haemorrhages, which are characteristic in both infections, are described. Apoptotic cell death is regarded as the predominant cause of lymphopenia in both virus infections.
Asunto(s)
Virus de la Fiebre Porcina Africana/patogenicidad , Fiebre Porcina Africana/virología , Virus de la Fiebre Porcina Clásica/patogenicidad , Peste Porcina Clásica/virología , Macrófagos/virología , Monocitos/virología , Fiebre Porcina Africana/etiología , Animales , Apoptosis , Peste Porcina Clásica/etiología , Citocinas/metabolismo , Macrófagos/inmunología , Monocitos/inmunología , PorcinosRESUMEN
Twenty pigs were inoculated with a virulent isolate (Quillota strain) of classical swine fever (CSF) virus to determine the chronological development of lesions in bone marrow. Histopathologic, ultrastructural and immunohistochemical (detection of viral antigen gp55, myeloid-histiocyte antigen, CD3 antigen, and FVIII-rag), and morphometric techniques were employed. Viral antigen was detected from 2 days postinfection (dpi) in stromal and haematopoitic cells, and severe atrophy related to apoptosis of haematopoitic cells was observed. Megakaryocytes (MKs) did not show significant changes in number, but there were important qualitative changes including 1) increased numbers of cloud-nuclei MKs, microMKs, apoptotic MKs, and atypical nucleated MKs and 2) decreased number of typical nucleated MKs. Morphometric study of these cells showed a decrease in cytoplasmic area. MK infection was detected from 2 dpi, but in a small percentage of cells. Myeloid cells showed quantitative changes, with an increase in granulocyte numbers. Apoptosis of lymphocytes and viral infection of erythroblasts were also observed. The main changes in stroma were depletion of T lymphocytes in the middle phase of the experiment and macrophages. Viral infection was also observed in these cells. MK lesions suggest dysmegakaryocytopoiesis, which would aggravate the thrombocytopenia already present and could be responsible for it. Granulocyte changes would lead to the appearance of circulating immature forms, whereas lymphocyte apoptosis in bone marrow would contribute to lymphopenia.
Asunto(s)
Médula Ósea/patología , Virus de la Fiebre Porcina Clásica/crecimiento & desarrollo , Peste Porcina Clásica/patología , Animales , Antígenos Virales/metabolismo , Apoptosis , Médula Ósea/ultraestructura , Médula Ósea/virología , Complejo CD3/metabolismo , Peste Porcina Clásica/virología , Virus de la Fiebre Porcina Clásica/ultraestructura , Femenino , Granulocitos/virología , Inmunohistoquímica/veterinaria , Recuento de Leucocitos/veterinaria , Masculino , Megacariocitos/virología , Microscopía Electrónica/veterinaria , Recuento de Plaquetas/veterinaria , Porcinos , Factor de von Willebrand/metabolismoRESUMEN
To gain further insight into the pathogenesis of African swine fever (ASF), the cytokine expression by macrophages in spleen and lymph nodes were examined. Twenty-one piglets were inoculated with the highly virulent isolate Spain-70 of ASF virus and killed in groups at 1-7 post-inoculation days (pid). An increase in the immunohistochemical detection of proinflammatory monokines in spleen and renal and gastrohepatic lymph nodes is reported, along with an increase in the serum levels of TNF-alpha and IL-1 beta. The expression of these cytokines is detected simultaneously in time and space with the viral protein 73 (vp 73) of the ASF virus detection. Our results demonstrate that mononuclear phagocyte system cell activation results in the release of several cytokines that could induce apoptosis of lymphocytes and haemodynamic changes.
Asunto(s)
Fiebre Porcina Africana/inmunología , Interleucinas/biosíntesis , Ganglios Linfáticos/inmunología , Macrófagos/metabolismo , Monocinas/biosíntesis , Bazo/inmunología , Factor de Necrosis Tumoral alfa/biosíntesis , Fiebre Porcina Africana/virología , Virus de la Fiebre Porcina Africana/fisiología , Animales , Femenino , Inmunohistoquímica , Ganglios Linfáticos/citología , Activación de Macrófagos , Masculino , Bazo/citología , Porcinos , Factores de TiempoRESUMEN
Twenty pigs were inoculated with a virulent classical swine fever virus isolate to determine the mechanism responsible for thrombocytopenia using histopathologic, ultrastructural, and immunohistochemical (detection of viral antigens gp55 and FVIII-rag) techniques. In animals euthanatized at 2, 4, and 6 days postinoculation (dpi), clusters of granular material staining positive for FVIII-rag were observed in splenic cords, the marginal zone, hepatic sinusoids, and the perisinusoidal space. Moreover, numerous macrophages in these areas were swollen and displayed an intensely positive granular and cytoplasmic reaction. Cell alterations indicative of platelet activation and secretory and phagocytic activation of resident macrophages were also observed in these sites at 2 and 4 dpi. These results suggest that the thrombocytopenia observed in pigs is caused in the first instance by massive activation and subsequent phagocytosis of platelets secondary to the release of platelet-activating factors by activated macrophages.
Asunto(s)
Peste Porcina Clásica/patología , Agregación Plaquetaria , Enfermedades de los Porcinos/patología , Trombocitopenia/veterinaria , Animales , Capilares/patología , Femenino , Masculino , Tamaño de los Órganos , Recuento de Plaquetas/veterinaria , Valores de Referencia , Bazo/patología , Bazo/ultraestructura , Porcinos , Trombocitopenia/etiología , Trombocitopenia/patologíaRESUMEN
Classical swine fever (CSF) was induced in 20 pigs by inoculation with a virulent strain of CSF virus to determine sequential changes (2, 4, 7, 10 and 14 days post-inoculation) in the number and morphology of splenic macrophages (red pulp and lymphoid marginal zone) and thus to assess the role of these cells in the pathogenesis of the disease. The first splenic cells to be infected with CSF virus were macrophages in the marginal zone followed by other macrophage populations. The initial phase of CSF was associated with an increase in splenic macrophage numbers in the marginal zone and a decrease in the red pulp. Subsequently, the numbers in the red pulp increased. The study suggested that infection, mobilization and apoptosis of splenic macrophages play an important role in the spread of CSF virus in vivo. Moreover, the secretory changes that occurred in macrophages in the initial phase of the infection suggested that macrophages release chemical mediators capable of modulating pathogenesis.
Asunto(s)
Virus de la Fiebre Porcina Clásica/fisiología , Peste Porcina Clásica/patología , Macrófagos/patología , Bazo/patología , Animales , Antígenos Virales/inmunología , Biomarcadores/análisis , Recuento de Células/veterinaria , Peste Porcina Clásica/etiología , Peste Porcina Clásica/metabolismo , Virus de la Fiebre Porcina Clásica/aislamiento & purificación , Virus de la Fiebre Porcina Clásica/ultraestructura , Vesículas Citoplasmáticas/ultraestructura , Femenino , Técnicas para Inmunoenzimas/veterinaria , Cuerpos de Inclusión Viral/ultraestructura , Macrófagos/metabolismo , Masculino , Factores de TiempoRESUMEN
Twenty-six pigs were inoculated with a virulent isolate (Quillota strain) of classical swine fever (hog cholera) virus to determine the chronological development of lesions in the renal glomeruli and the pathogenesis of glomerular damage and immunocomplex deposition. The study included the use of histopathological, ultrastructural and immunohistochemical (detection of viral antigen gp55, myeloid-histiocyte antigen, IgM, IgG and C1q) techniques. The main changes in glomerular structure were observed from 7 days post-inoculation (dpi) onwards, at which time the glomeruli showed macrophage infiltrations in the mesangium, and viral infection in circulating cells, glomerular endothelial cells and podocytes. Moreover, significant subcellular changes were detected in podocytes, which appeared swollen, with fusion of foot processes. Immunocomplex deposits immunoreactive for IgM, IgG and C1q were detected in mesangial, subepithelial and subendothelial areas from 10 dpi, but viral antigen was not detected as a component of these deposits; fusion of foot processes had increased in severity, especially near immunocomplex deposits. All these changes had increased still further in the final phase of the experiment (14 dpi), with neutrophil infiltrations in the mesangium.
Asunto(s)
Complejo Antígeno-Anticuerpo/inmunología , Peste Porcina Clásica/inmunología , Glomerulonefritis/inmunología , Glomérulos Renales/inmunología , Porcinos , Animales , Peste Porcina Clásica/patología , Peste Porcina Clásica/virología , Virus de la Fiebre Porcina Clásica/inmunología , Virus de la Fiebre Porcina Clásica/aislamiento & purificación , Virus de la Fiebre Porcina Clásica/fisiología , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Mesangio Glomerular/inmunología , Mesangio Glomerular/ultraestructura , Mesangio Glomerular/virología , Glomerulonefritis/patología , Glomerulonefritis/virología , Técnicas para Inmunoenzimas/veterinaria , Inmunoglobulina M/análisis , Glomérulos Renales/patología , Glomérulos Renales/virología , Masculino , Microscopía Electrónica/veterinariaRESUMEN
To gain further insight into the pathogenesis of classical swine fever (CSF), the changes induced by hog cholera (HC) virus in pulmonary intravascular macrophages (PIMs) were examined. Twelve pigs were inoculated by the intramuscular route with a virulent strain of HC virus (Quillota strain) and killed in groups of three at 4, 7, 10 and 14 days post-inoculation. Immunohistochemical and ultrastructural examination revealed HC virus infection in endothelial cells, PIMs, and interstitial and alveolar macrophages. In addition to viral replication, a predominant feature was the secretory activation of PIMs, characterized by expanded rough endoplasmic reticulum and hyperplastic Golgi complexes. The results obtained suggest that macrophage activation and the subsequent release of pro-inflammatory mediators play an important role in the pathogenesis of CSF.
Asunto(s)
Virus de la Fiebre Porcina Clásica/fisiología , Peste Porcina Clásica/patología , Pulmón/patología , Macrófagos Alveolares/patología , Animales , Recuento de Células , Peste Porcina Clásica/etiología , Virus de la Fiebre Porcina Clásica/ultraestructura , Endotelio Vascular/patología , Endotelio Vascular/virología , Femenino , Técnicas para Inmunoenzimas/veterinaria , Pulmón/irrigación sanguínea , Pulmón/virología , Macrófagos Alveolares/virología , Masculino , Microscopía Electrónica/veterinaria , Orgánulos/ultraestructura , Porcinos , Proteínas del Envoltorio Viral/análisisRESUMEN
To study the effect of hog cholera virus on the epithelial cells of the bronchiolar mucosa, 12 pigs were inoculated with a highly virulent strain. Immunohistochemical and ultrastructural examination of the ciliated epithelial cells demonstrated an increase in the number of atypical cilia. The latter showed alterations in the microtubular pattern, possibly resulting from viral interference with the normal metabolism of the epithelial cells.
Asunto(s)
Bronquios/ultraestructura , Cilios/ultraestructura , Virus de la Fiebre Porcina Clásica/fisiología , Peste Porcina Clásica/patología , Animales , Bronquios/virología , Virus de la Fiebre Porcina Clásica/inmunología , Virus de la Fiebre Porcina Clásica/ultraestructura , Epitelio/ultraestructura , Epitelio/virología , Femenino , Técnicas para Inmunoenzimas , Masculino , Microscopía Electrónica , Mucosa Respiratoria/ultraestructura , Mucosa Respiratoria/virología , Porcinos , Proteínas del Envoltorio Viral/análisis , Replicación ViralRESUMEN
Monokines are glycoproteins, synthesised by macrophages, which exert various effects on the organism. The most important monokines are interleukin (IL)-1alpha, IL-1beta, tumor necrosis factor (TNF)-alpha and IL-6. This paper reports on immunohistochemical techniques developed for the detection of IL-alpha, IL-1beta, IL-6 and TNF-alpha in fixed and paraffin-embedded pig tissues (spleen, lymph nodes, thymus, liver and kidney). Different fixatives (buffered formalin, acetic formalin, paraformadehyde-lysine-periodate and Bouin solution), and antigen unmasking techniques (permeabilisation with Tween 20, pronase enzymatic digestion and microwave-citrate buffer) were used. We describe different protocols for detection of monokines using polyclonal antibodies against the studied monokines. No signal was obtained with monoclonal antibodies against pig-TNF-alpha and human IL-1alpha. Bouin solution was shown to be the best fixative for immunohistochemical detection of IL-1alpha, TNF-alpha, and IL-6, using permeabilisation with Tween 20 as an unmasking antigen method. Acetic formalin was shown to be the best fixative for IL-1beta detection, not needing antigen retrieval techniques. Macrophages were identified as the main cytokine-producing cells, although other types of cells also stained positively to some cytokines. These techniques represent valuable tools for studies of the pathogenesis of viral and bacterial diseases, and of the immune system of the pigs.
Asunto(s)
Anticuerpos , Inmunohistoquímica/veterinaria , Monocinas/aislamiento & purificación , Porcinos/inmunología , Animales , Inmunohistoquímica/métodos , Interleucina-1/inmunología , Interleucina-1/aislamiento & purificación , Interleucina-6/inmunología , Interleucina-6/aislamiento & purificación , Riñón/inmunología , Hígado/inmunología , Ganglios Linfáticos/inmunología , Monocinas/inmunología , Bazo/inmunología , Timo/inmunología , Factor de Necrosis Tumoral alfa/inmunología , Factor de Necrosis Tumoral alfa/aislamiento & purificaciónRESUMEN
Thirty pigs were inoculated with a virulent isolate (Quillota strain) of classical swine fever (hog cholera) virus to establish the chronological occurrence of lesions in the kidney and to determine the mechanism responsible for renal haemorrhages. The study included the use of histopathological, ultrastructural, immunohistochemical (detection of viral antigen gp55, MAC387, lambda chains, CD3 and C1q) and morphometrical techniques (vascular area). Renal interstitial oedema and haemorrhages were detected from 7 days post-inoculation (dpi), associated with a slight interstitial mononuclear infiltrate and evidence of viral infection in macrophages and fibroblasts, and in a small proportion of lymphocytes. Viral infection was not detected in capillary endothelial cells. An intense mononuclear infiltrate, with B cells, T cells and small numbers of macrophages, was detected from 10 dpi. In the final phase of the experiment (14 dpi), slight proliferation and degranulation of mast cells were observed. Increased expression of the C1q component of complement was also detected. A significant increase in vascular area was observed from 7 dpi. These results suggest that haemorrhages observed in the kidneys of pigs inoculated with the Quillota strain resulted from erythrodiapedesis and increased vascular permeability, probably aggravated by mast cell degranulation in the final stage of the experiment. The results suggested that mast cell degranulation was linked to activation of the complement system.
Asunto(s)
Movimiento Celular/fisiología , Virus de la Fiebre Porcina Clásica , Peste Porcina Clásica/etiología , Eritrocitos/fisiología , Hemorragia/etiología , Enfermedades Renales/etiología , Animales , Antígenos Virales/análisis , Biomarcadores/análisis , Capilares/metabolismo , Capilares/ultraestructura , Capilares/virología , Fragilidad Capilar/fisiología , Permeabilidad Capilar/fisiología , Degranulación de la Célula/fisiología , Peste Porcina Clásica/inmunología , Peste Porcina Clásica/patología , Virus de la Fiebre Porcina Clásica/inmunología , Vesículas Citoplasmáticas/fisiología , Vesículas Citoplasmáticas/ultraestructura , Edema/etiología , Edema/patología , Endotelio Vascular/metabolismo , Endotelio Vascular/ultraestructura , Endotelio Vascular/virología , Eritrocitos/citología , Femenino , Hemorragia/patología , Enfermedades Renales/patología , Masculino , Mastocitos/fisiología , Mastocitos/ultraestructura , PorcinosAsunto(s)
Infecciones por Chlamydia/veterinaria , Enfermedades de los Porcinos/microbiología , Síndrome Debilitante/veterinaria , Animales , Anticuerpos Antibacterianos/análisis , Infecciones por Chlamydia/complicaciones , Infecciones por Chlamydia/inmunología , Huésped Inmunocomprometido , Enfermedades Intestinales/microbiología , Enfermedades Intestinales/veterinaria , Porcinos , Enfermedades de los Porcinos/patología , Síndrome Debilitante/complicacionesRESUMEN
The presence of apoptotic cell death was evaluated in routinely processed tissue samples of 39 neoplasms of the skin and subcutaneous tissues of the dog using the method of terminal deoxynucleotidyl transferase (T d T) mediated deoxyuridine-5'-triphosphate (d UTP)-biotin nick end labelling (TUNEL). The degree of apoptosis was related to the frequency of mitosis, an index of cell proliferation. The correlation between the apoptotic index (AI), the percentage of positive cells after randomly enumerating 1000 cells and the mitotic count (MC), the number of mitotic figures in 10 fields at a magnification of 400 times was assessed by the Spearman non-parametric correlation test. TUNEL signals were observed in all types of tumours as brown products detected in non-pyknotic nuclei, in non-identifiable rounded structures (so-called apoptotic bodies) and occasionally in the cytoplasm, either singly or in combination. An inverse relationship between AI and MC was observed in benign tumours, while no correlation was found between AI and MC in either malignant or locally invasive tumours. Among benign tumours, intracutaneous cornifying epithelioma, fibroma, haemangioma and Schwannoma had high AI and low MC, while histiocytomas had low AI and high MC and pilomatrixomas low AI and MC. All malignant tumours had low AI and high MC, except for fibrosarcomas, which had high AI and MC. Finally, higher heterogeneity was observed among locally invasive tumours, as they had high AI and low MC (squamous cell carcinomas), and low AI with either low MC (haemangiopericytomas) or high MC (basal cell tumours). The classification of the tumours according to their AI (>15.8% high and <15.8% low) and MC (>9 high, <9 low) did not reflect the clinical behaviour of some tumour types.
Asunto(s)
Enfermedades de los Perros/patología , Neoplasias Cutáneas/veterinaria , Adenocarcinoma/patología , Adenocarcinoma/veterinaria , Adenoma/patología , Adenoma/veterinaria , Animales , Apoptosis , Carcinoma/patología , Carcinoma/veterinaria , Carcinoma Basocelular/patología , Carcinoma Basocelular/veterinaria , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/veterinaria , Enfermedades de los Perros/cirugía , Perros , Fibrosarcoma/patología , Fibrosarcoma/veterinaria , Histiocitoma Fibroso Benigno/patología , Histiocitoma Fibroso Benigno/veterinaria , Etiquetado Corte-Fin in Situ , Sarcoma de Mastocitos/patología , Sarcoma de Mastocitos/veterinaria , Mitosis , Invasividad Neoplásica , Neoplasias de las Glándulas Sebáceas/patología , Neoplasias de las Glándulas Sebáceas/veterinaria , Neoplasias Cutáneas/clasificación , Neoplasias Cutáneas/patología , Neoplasias Cutáneas/cirugíaRESUMEN
Twelve pigs were inoculated with a highly virulent strain of hog cholera virus (HCV) to study viral infection of megakaryocytes in the bone marrow and spleen. Immunohistochemical and ultrastructural examination revealed HCV infection in a small proportion (2.5-9.0%) of these cells from the 2nd to the 9th day after inoculation, at which time the experiment was terminated. Megakaryocyte infection accounts for the presence of viral antigens in platelets. The latter may represent a passive vehicle for spreading the virus in the animal.
Asunto(s)
Médula Ósea/patología , Virus de la Fiebre Porcina Clásica/fisiología , Virus de la Fiebre Porcina Clásica/ultraestructura , Peste Porcina Clásica/virología , Megacariocitos/virología , Bazo/patología , Animales , Antígenos Virales/análisis , Médula Ósea/virología , Recuento de Células , Peste Porcina Clásica/patología , Femenino , Técnicas para Inmunoenzimas/veterinaria , Cuerpos de Inclusión/ultraestructura , Masculino , Megacariocitos/ultraestructura , Recuento de Plaquetas/veterinaria , Bazo/virología , PorcinosRESUMEN
Interpretation of changes in bone marrow during infectious processes is quite complex. This paper reports bone marrow lesions observed in pigs inoculated with a moderately virulent ASF virus strain and studies their relationship to the pathogenesis of the disease. In this work, we have carried out the structural and ultrastructural study of the bone marrow of 14 Large White x Landrace pigs that were inoculated by the intramuscular route with 10(5) 50% hemodsorbing doses (HAD50) of the Dominican Republic'78 ASF virus strain. The inoculated pigs were killed at 3, 5, 7, 9, 11, 13, 15 and 17 days postinjection. Analysis of cells and structures belonging to the two main bone-marrow compartments, the hematopoietic cells and the hematopoietic micro-environment, showed that after inoculation with a moderately virulent strain, the most significant changes occurred in macrophages and megakaryocytes, consisting in virus replication in these cell populations and apoptosis of megakaryocytes, related with the sudden and transitory thrombocytopenia detected in the subacute ASF.
Asunto(s)
Fiebre Porcina Africana/patología , Células de la Médula Ósea/ultraestructura , Médula Ósea/ultraestructura , Fiebre Porcina Africana/virología , Animales , Médula Ósea/patología , Médula Ósea/virología , Células de la Médula Ósea/patología , Células de la Médula Ósea/virología , Hematopoyesis , Células Madre Hematopoyéticas , Porcinos , VirulenciaRESUMEN
Formalin-fixed, paraffin embedded tissues obtained from 40 pigs inoculated with a field isolate of hog cholera virus were examined for the presence of Gp55, a major structural protein of the virus envelope, using a monoclonal antibody-based immunohistochemical test with the avidin-biotin-peroxidase complex method. Immunoreactivity was detected in hog cholera virus-infected tissues but not in control pigs tissues, African swine fever virus-infected tissues, or bovine viral diarrhea virus-infected porcine or bovine tissues. The first positive reactions were seen in lymphatic tissues, digestive tract and skin on postinoculation day (pid) 4, respiratory and urinary tissues on pid 5, nervous tissues on pid 6, and endocrine tissues on pid 7. These staining reactions persisted until the last observation on pid 18. Hog cholera virus antigen was not detected in heart tissue at any time. The highest levels of antigen detection were found in tonsils, spleen, and pancreas, although the esophageal mucosa and skin epithelial cells were also intensely and widely stained. The cellular staining pattern of Gp55 had a ubiquitous distribution. It was found in epithelial cells, macrophages and circulating monocytes, endothelial cells, lymphoid cells, and glial cells. The results showed a high specificity and high sensitivity for detecting hog cholera Gp55 in formalin-fixed, paraffin embedded tissue samples. This method allows precise association of Gp55 with specific cells, tissues, and histologic lesions, making the technique suitable for use in routine diagnosis of hog cholera.
Asunto(s)
Virus de la Fiebre Porcina Clásica/aislamiento & purificación , Peste Porcina Clásica/patología , Proteínas del Envoltorio Viral/análisis , Animales , Encéfalo/patología , Encéfalo/virología , Bovinos , Diagnóstico Diferencial , Sistema Digestivo/patología , Sistema Digestivo/virología , Inmunohistoquímica/métodos , Tejido Linfoide/patología , Tejido Linfoide/virología , Valores de Referencia , Sistema Respiratorio/patología , Sistema Respiratorio/virología , Piel/patología , Piel/virología , Porcinos , Sistema Urinario/patología , Sistema Urinario/virologíaRESUMEN
This paper describes the clinical and histopathological features of an outbreak of cutaneous squamous cell carcinomas in a flock of Berrichon du Cher sheep in south-western Spain. The outbreak was unusual because of the high prevalence recorded (15 to 18 per cent over the last two years) and because the majority of the lesions involved the eyelids. Adult and old sheep were the most commonly affected, and the largest numbers of cases were recorded in summer and autumn. The initial lesions were non-specific and consisted of erythema, hyperkeratosis, actinic keratosis, periocular dermatitis and conjunctivitis. The tumours grew slowly but progressively over a period of one to two years, with frequent complications by bacterial infections or secondary myiasis. Histopathological examination of the tumours revealed well-differentiated squamous cell carcinomas, surrounded by a moderate to abundant infiltrate of lymphocytes, plasma cells and macrophages. The overlying epidermis was frequently ulcerated and inflamed. The possible relationships between the high prevalence of the condition and the poorly pigmented skin of this breed, the extensive farming system employed and the exposure of the animals to ultraviolet radiation were analysed.