African swine fever. I. Morphological changes and virus replication in blood platelets of pigs infected with virulent haemadsorbing and non-haemadsorbing isolates

Replicating and mature viral particles were detected with the transmission electron microscope in blood platelets of pigs infected with virulent haemadsorbing and non-haemadsorbing African swine fever virus isolates. Although platelet numbers decreased terminally in infected pigs, the most noticeable morphological damage to these cells apparent in the last 2 days of the disease included cytoplasmic swelling, vacuolation, fragmentation and loss of dense granules.     

Neutralization of African swine fever virus by sera from African swine fever-resistant pigs

Sera from African swine fever-resistant pigs with infection-inhibitory activity decreased virus replication in infected porcine buffy coat cultures. This same effect was observed even after virus was adsorbed. The infection-inhibition was not reversed by removing the immune serum from the assay cultures. Reduction of African swine fever virus replication by immune sera was demonstrated by fluorescent focus assay on MS cell line cultures. Virus-neutralization tests showed a persistent fraction of non-neutralized virus, which was not demonstrable by infection-inhibition tests. One hypothesis for explaining this difference is proposed.     

African swine fever. II. Functional disturbances of thrombocytes in pigs infected with virulent haemadsorbing and non-haemadsorbing virus isolates

Increased bleeding time, impaired blood clot retraction and decreased thrombocyte aggregation were observed in pigs infected with virulent haemadsorbing and non-haemadsorbing African swine fever virus isolates. These changes appeared to be more frequent and more severe in pigs infected with the haemadsorbing virus isolates than in those infected with the non-haemadsorbing isolate. Moreover, the onset and severity of these changes followed the numerical decrease and morphological damage to these cells apparent in the last 2-3 days of the disease.     

我国流行的非洲猪瘟病毒:高致病力,强传播能力

非洲猪瘟(African Swine Fever,ASF)是一种急性、烈性、病毒性传染病,主要感染家猪和野猪,其发病率和死亡率极高,接近100%,被世界动物卫生组织(OIE)列为必须报告的动物疫病,被我国列为一类动物疫病。当前该病缺乏有效的防控用疫苗及药物,主要通过捕杀发病动物及严格的生物安全措施来控制。该病病原是非洲猪瘟病毒(African Swine Fever Virus,ASFV),属于非洲猪瘟病毒科(Asfarviridae)非洲猪瘟病毒属(Asfivirus)唯一成员,其基因组长约170 kb^193 kb,为双链DNA,编码151~167个病毒蛋白。     

Prevalence of African swine fever virus and classical swine fever virus antibodies in pigs in Benue State,Nigeria

Tropical Animal Health and Production - This study investigated the prevalence of African swine fever virus (ASFV) and classical swine fever virus (CSFV) antibodies in pigs in Benue State, Nigeria....     

Abortion in sows experimentally infected with African swine fever virus: pathogenesis studies

Thirteen sows that were 38 to 92 days pregnant were experimentally infected with an African swine fever (ASF) virus strain of low virulence (Dominican Republic isolate). Seven of 11 sows that were not killed had aborted. The pathogenesis of the abortions was studied, using virus isolation, tissue immunofluoresence, and histopathologic techniques. African swine fever virus was recovered from 179 of 1,329 (13.5%) fetal tissues tested. The 3 fetal tissues most frequently yielding virus were the fetal placenta, amniotic fluid, and fetal heart blood. Virus was not recovered from fetal tissues obtained from 2 of the aborting sows. Direct immunofluorescent microscopy for ASF viral antigen was done on approximately 1,175 fetal tissues. Although brightly fluorescing cells were common in maternal tissues, specific immunofluorescence was present in only placental tissues from 2 sows. Microscopic lesions in fetal tissues were inconsistent and included mild focal placentitis, mild heptic degeneration and necrosis, and mild interstitial pneumonia. These changes were not considered to be sufficiently specific to have diagnostic significance. In marked contrast to these changes in the fetal tissues, maternal tissues had high titers of virus, with marked necrosis of lymphoid tissues, and contained many cells with ASF viral antigen. We conclude that specific diagnosis of abortion resulting from ASF infection should, therefore, be based on examination of maternal tissues, rather than fetal tissues. The pregnancy failure seems to result from the effects of the virus infection on the dam more so than from direct viral damage to the placenta or fetus.     

Dynamics of African swine fever virus shedding and excretion in domestic pigs infected by intramuscular inoculation and contact transmission

African swine fever virus (ASFV) is a highly virulent swine pathogen that has spread across Eastern Europe since 2007 and for which there is no effective vaccine or treatment available. The dynamics of shedding and excretion is not well known for this currently circulating ASFV strain. Therefore, susceptible pigs were exposed to pigs intramuscularly infected with the Georgia 2007/1 ASFV strain to measure those dynamics through within- and between-pen transmission scenarios. Blood, oral, nasal and rectal fluid samples were tested for the presence of ASFV by virus titration (VT) and quantitative real-time polymerase chain reaction (qPCR). Serum was tested for the presence of ASFV-specific antibodies. Both intramuscular inoculation and contact transmission resulted in development of acute disease in all pigs although the experiments indicated that the pathogenesis of the disease might be different, depending on the route of infection. Infectious ASFV was first isolated in blood among the inoculated pigs by day 3, and then chronologically among the direct and indirect contact pigs, by day 10 and 13, respectively. Close to the onset of clinical signs, higher ASFV titres were found in blood compared with nasal and rectal fluid samples among all pigs. No infectious ASFV was isolated in oral fluid samples although ASFV genome copies were detected. Only one animal developed antibodies starting after 12 days post-inoculation. The results provide quantitative data on shedding and excretion of the Georgia 2007/1 ASFV strain among domestic pigs and suggest a limited potential of this isolate to cause persistent infection.     

Localization of classical swine fever virus from chronically infected pigs by in situ hybridization and immunohistochemistry

Classical swine fever (CSF) virus (CSFV) nucleic acid and antigen were detected in 15 pigs with naturally occurring chronic CSF by in situ hybridization and immunohistochemistry. The most consistent and prominent microscopic lesions were perivascular mononuclear cell infiltration and gliosis in the central nervous system of pigs with chronic CSF. Positive cells typically exhibited a dark brown (in situ hybridization) or red (immunohistochemistry) reaction product in the cytoplasm without background staining. A positive signal for both in situ hybridization and immunohistochemistry was detected in mononuclear cells and lymphocytes of lymphoid tissues. Viral nucleic acid was detected in some tissue sections in the absence of viral antigen. The in situ hybridization technique developed in this study was useful for the detection of CSFV RNA in tissues taken from chronically infected pigs and may be a valuable technique for studying the pathogenesis of chronic CSFV infection.     

Classical swine fever virus in plasma and peripheral blood mononuclear cells of acutely infected swine

The distribution of classical swine fever virus (CSFV) in plasma, monocytes, T and B lymphocytes in peripheral blood was monitored during experimentally induced acute classical swine fever infection in piglets. Six piglets were infected with 10(3.8) TCID50 of virus and blood samples taken up to 18 days post-inoculation (p.i.). Infectious virus was detected in monocytes, T and B lymphocytes to similar titres in five of the six infected piglets. Infectious virus was detected earlier in plasma than in any of the mononuclear cell subpopulations. No significant difference was observed in the period of time in which virus could be isolated from the three cell subpopulations. While a progressive lymphopenia developed, a marked B cell depletion was observed. However, B cells were apparently replaced by non-IgM-bearing mononuclear cells, as the proportion 'total lymphocyte/total leucocytes' remained unaltered throughout the experiment. Virus titres in plasma and peripheral blood mononuclear cells showed a tendency to increase as the disease progressed to its outcome.     

An immunohistochemical study of the tonsils in pigs with acute African swine fever virus infection

An immunohistochemical study of the tonsils was carried out to gain further insight in the pathogenesis of acute African swine fever (ASF). Twenty-one pigs were inoculated by intramuscular route with a highly virulent isolate of ASF virus and painlessly killed at 1-7dpi. Viral antigen was highly distributed in the tonsil from 3 to 4dpi and an increase in the number of monocyte-macrophages was very evident at the same days post inoculation. This phenomenon was observed together with an increase of the expression of proinflammatory cytokines (Tumour necrosis factor alpha and Interleukin-1 alpha) and the apoptosis of lymphocytes studied by the terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling (TUNEL) technique and haemorrhages. With these results, we can conclude that the tonsil is suffering similar lesions than those observed in other lymphoid organs in acute African swine fever, even when the route of inoculation is the intramuscular and not oral-nasal.     

Interferon status and white blood cells during infection with African swine fever virus in vivo

African swine fever virus (ASFV) is the causative agent of African swine fever that is the significant disease of domestic pigs, with high rates of mortality. ASFV is double-stranded DNA virus whose genes encode some proteins that are implicated in the suppression of host immune response. In this study, we have modeled in vivo infection of ASFV for determination of interferon (IFN) status in infected pigs. We measured the level of IFN-α, -β and -γ by enzyme-linked immunosorbent assay and showed that the level of IFN-α sharply decreased during infection. Unlike IFN-α, the level of IFN-β and -γ increased from the 2nd and 4th days post-infection, respectively. Also, we analyzed the population dynamics of peripheral white blood cells of infected pigs due to their important role in host immune system. We showed that the atypical lymphocytes appeared after short time of infection and this result is in accordance with our previous study done in vitro. At the last day of infection about 50% of the total white blood cells were destroyed, and the remaining cells were represented mainly by small-sized lymphocytes, reactive lymphocytes and lymphoblasts.     

Detection of antibodies against African swine fever virus using infected monolayers and monoclonal antibodies

Three monoclonal antibodies, specific for porcine IgG, IgM and IgA, were used to develop isotype-specific immunoperoxidase monolayer assays for the detection of antibodies against African swine fever virus. A mixture of anti-IgM and anti-IgG monoclonal antibodies was used in an assay designed for screening sera. This test was compared with a commercially available ELISA by using experimental sera and field sera obtained after an outbreak of African swine fever on two farms in the Netherlands in 1986. Although the ELISA was less sensitive than the immunoperoxidase monolayer assay on sera taken early after infection, the tests were equally useful for screening purposes. The isotype-specific assays gave epizootiological information about the stage of infection on the two farms.     

The formation process of button ulcers in pigs experimentally infected with a subgenotype 2.1 isolate of classical swine fever virus

We evaluated the role of classical swine fever virus (CSFV) in the formation of button ulcers in the mucosa of the gastrointestinal tract. Histopathological and immunohistochemical analyses of pigs experimentally infected with a subgenotype 2.1 isolate of CSFV, which was isolated in Japan in 2019, revealed follicular necrosis in the submucosal mucosa-associated lymphoid tissue and herniation of crypts as factors that contribute to the development of button ulcers during CSFV infection. These findings indicate that CSFV induces follicular necrosis and is one of the causative agents of button ulcers in pigs.     

Detection and quantification of classical swine fever virus in air samples originating from infected pigs and experimentally produced aerosols

During epidemics of classical swine fever (CSF), neighbourhood infections occurred where none of the 'traditional' routes of transmission like direct animal contact, swill feeding, transport contact or transmission by people could be identified. A hypothesized route of virus introduction for these herds was airborne transmission. In order to better understand this possible transmission route, we developed a method to detect and quantify classical swine fever virus (CSFV) in air samples using gelatine filters. The air samples were collected from CSFV-infected pigs after experimental aerosolization of the virus. Furthermore, we studied the viability of the virus with time in aerosolized state. Three strains of CSFV were aerosolized in an empty isolator and air samples were taken at different time intervals. The virus remained infective in aerosolized state for at least 30 min with half-life time values ranging from 4.5 to 15 min. During animal experiments, concentrations of 10(0.3)-10(1.6)TCID(50)/m(3) CSFV were detected in air samples originating from the air of the pig cages and 10(0.4)-10(4.0)TCID(50)/m(3) from the expired air of infected animals. This is the first study describing the isolation and quantification of CSFV from air samples originating from infected pigs and their cages, supporting previous findings that airborne transmission of CSF is feasible.