RESUMO
African swine fever (ASF) is a contagious viral hemorrhagic disease of domestic pigs and wild boars. The disease is notifiable to the World Organisation for Animal Health (OIE) and is responsible for high mortality and serious economic losses. PCR and real-time PCR (qPCR) are the OIE-recommended standard methods for the direct detection of African swine fever virus (ASFV) DNA. The aim of our work was the simplification and standardization of the molecular diagnostic workflow in the lab. For validation of this "easy lab" workflow, different sample materials from animal trials were collected and analyzed (EDTA blood, serum, oral swabs, chewing ropes, and tissue samples) to identify the optimal sample material for diagnostics in live animals. Based on our data, the EDTA blood samples or bloody tissue samples represent the best specimens for ASFV detection in the early and late phases of infection. The application of prefilled ready-to-use reagents for nucleic acid extraction or the use of a Tissue Lysis Reagent (TLR) delivers simple and reliable alternatives for the release of the ASFV nucleic acids. For the qPCR detection of ASFV, different published and commercial kits were compared. Here, a lyophilized commercial kit shows the best results mainly based on the increased template input. The good results of the "easy lab" strategy could be confirmed by the ASFV detection in field samples from wild boars collected from the 2020 ASFV outbreak in Germany. Appropriate internal control systems for extraction and PCR are key features of the "easy lab" concept and reduce the risk of false-negative and false-positive results. In addition, the use of easy-to-handle machines and software reduces training efforts and the misinterpretation of results. The PCR diagnostics based on the "easy lab" strategy can realize a high sensitivity and specificity comparable to the standard PCR methods and should be especially usable for labs with limited experiences and resources.
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Vírus da Febre Suína Africana/isolamento & purificação , Febre Suína Africana/diagnóstico , DNA Viral/genética , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sus scrofa/virologia , Suínos/virologia , Febre Suína Africana/sangue , Febre Suína Africana/epidemiologia , Febre Suína Africana/virologia , Vírus da Febre Suína Africana/genética , Animais , DNA Viral/isolamento & purificação , Surtos de Doenças/veterinária , Alemanha , Padrões de Referência , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Effective vaccines against porcine reproductive and respiratory syndrome virus (PRRSV), especially against highly pathogenic (HP) PRRSV are still missing. The objective of this study was to evaluate the protective efficacy of an experimental live attenuated PRRSV 2 vaccine, composed of two strains, against heterologous challenge with a Vietnamese HP PRRSV 2 field strain. For this reason, 20 PRRSV negative piglets were divided into two groups. The pigs of group 1 were vaccinated with the experimental vaccine, group 2 remained unvaccinated. All study piglets received an intranasal challenge of the HP PRRSV 2 on day 0 of the study (42 days after vaccination). Blood samples were taken on days 7 and 21 after vaccination and on several days after challenge. On day 28 after challenge, all piglets were euthanized and pathologically examined. RESULTS: On days 7 and 21 after vaccination, a PRRSV 2 viraemia was seen in all piglets of group 1 which remained detectable in seven piglets up to 42 days after vaccination. On day 3 after challenge, all piglets from both groups were positive in PRRSV 2 RT-qPCR. From day 7 onwards, viral load and number of PRRSV 2 positive pigs were lower in group 1 than in group 2. All pigs of group 1 seroconverted after PRRSV 2 vaccination. PRRSV antibodies were detected in serum of all study pigs from both groups from day 14 after challenge onwards. In group 2, moderate respiratory symptoms with occasional coughing were seen following the challenge with HP PRRSV 2. Pigs of group 1 remained clinically unaffected. Interstitial pneumonia was found in four piglets of group 1 and in all ten piglets of group 2. Histopathological findings were more severe in group 2. CONCLUSIONS: It was thus concluded that the used PRRSV 2 live experimental vaccine provided protection from clinical disease and marked reduction of histopathological findings and viral load in pigs challenged with a Vietnamese HP PRRSV 2 field strain.
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Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Vacinas Virais/uso terapêutico , Animais , Ensaio de Imunoadsorção Enzimática/veterinária , Masculino , Síndrome Respiratória e Reprodutiva Suína/imunologia , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Análise de Sequência de DNA/veterinária , Suínos/imunologia , Suínos/virologia , Resultado do Tratamento , Vacinas Atenuadas/uso terapêutico , Vacinas Virais/imunologiaRESUMO
BACKGROUND: In this study, six enzyme-linked immunosorbent assays (ELISA), intended for routine porcine reproductive and respiratory syndrome virus (PRRSV) herd monitoring, are tested for their ability to detect PRRSV specific antibodies in the serum of pigs after vaccination with an inactivated PRRSV type 1 vaccine and subsequent infection with a highly pathogenic (HP) PRRSV field strain. For this reason, ten piglets (group V) from a PRRSV negative herd were vaccinated twice at the age of 2 and 4 weeks with an inactivated PRRSV vaccine. Ten additional piglets (group N) from the same herd remained unvaccinated. Three weeks after second vaccination, each of the piglets received an intradermal application of an HP PRRSV field strain. Serum samples were taken before first vaccination as well as before and 3, 7, 10 and 14 days after HP PRRSV application. All serum samples were tested for PRRSV RNA by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) as well as for PRRSV antibodies with all six study ELISAs. RESULTS: At the beginning of the study (before vaccination), all of the piglets were PRRSV antibody negative with all study ELISAs. They also tested negative for PRRSV RNA measured by RT-qPCR. From day 3 after HP PRRSV application until the end of the study, a viremia was detected by RT-qPCR in all of the piglets. On day 0 (day of HP PRRSV application), nine out of ten piglets of the pre-vaccinated group tested PRRSV antibody positive with one of the tested ELISAs, although with lower S/P values than after infection. On day 10 after HP PRRSV application, all study ELISAs except one had significantly higher S/P or OD values, respectively more positive samples, in group V than in group N. CONCLUSIONS: Only one of the tested ELISAs was able to detect reliably PRRSV antibodies in pigs vaccinated with an inactivated PRRSV vaccine. With most of the tested ELISAs, higher S/P values respectively more positive samples after PRRSV infection were seen in the pre-vaccinated group than in the non-vaccinated.
Assuntos
Anticorpos Antivirais/sangue , Ensaio de Imunoadsorção Enzimática/veterinária , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/imunologia , Suínos , Vacinas de Produtos Inativados/imunologiaRESUMO
BACKGROUND: Porcine epidemic diarrhea (PED) is a syndrome that is characterized by rapidly spreading watery diarrhea affecting pigs of all ages, but with major effects on suckling piglets. The disease, as well as the causative Alphacoronavirus, the Porcine epidemic diarrhea virus (PEDV), was first described in Europe in the 1970s and since then has spread over many Asian and American countries, where it recently led to devastating effects on swine health and pork industry. While the disease was seldom reported in Europe within the last few decades, a few recent reports re-emergence of PED in German pig farms. The hitherto isolated German strain seems to be closely related to a low pathogenic PEDV variant from the USA. This case report describes the first detection of PEDV in Austria. CASE PRESENTATION: Reduced feed uptake and occasional diarrhea were observed in December 2014 in a group of fattening pigs, kept on an Austrian swine farm. The concerned pigs had been recently purchased from Germany. Within a few weeks, diarrhea became apparent also in pigs of Austrian origin, which were kept in a different stable on the same farm. Gastrointestinal symptoms among fattening pigs were generally mild, quickly resolving and did not lead to death. PEDV RNA was identified by RT-qPCR in pooled feces and serum and PEDV antibodies were detectable in serum in both groups of pigs. Phylogenetic analysis of the nearly complete PEDV spike gene shows that the Austrian PEDV strain is highly similar to other strains involved in recent outbreaks in Western and Central Europe. CONCLUSION: This is the first report demonstrating the presence of PEDV in Austria. The virus was probably introduced by purchasing piglets from a German source, which underlines the significance of trans-boundary animal trade for the distribution of highly contagious diseases, such as PED.
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Infecções por Coronavirus/veterinária , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , Animais , Áustria/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Epidemiologia Molecular , Vírus da Diarreia Epidêmica Suína/genética , SuínosRESUMO
The intracellular protozoan Toxoplasma gondii is distributed worldwide and infects many species of warm-blooded animals. Most mammals, including humans, can serve as intermediate hosts. This pathogen, with its zoonotic potential, causes toxoplasmosis, a condition that can range from subclinical to fatal in humans. It is therefore important to assess the occurrence of the pathogen, even if only indirectly through the detection of antibodies. Epidemiological data on the seroprevalence in wild animals, including invasive species, are rare in Poland. Therefore, we tested 197 wild raccoons (Procyon lotor) and 89 raccoon dogs (Nyctereutes procyonoides) from Zgorzelec County, southwestern Poland, for the presence of antibodies. Samples were collected between January 2019 and December 2020 and analysed using a commercial indirect modified agglutination test (MAT, cut-off 1:25). The statistical analysis revealed significant differences in seroprevalence between the two predatory species. Of the 197 surveyed raccoons, 96 (48.73%; 95% confidence interval (CI): 41.73-55.73%) tested positive, while 25 of the 89 raccoon dogs (28.09%; 95% CI: 18.70-37.48%) were positive. Regarding risk factors, body weight and sex influenced the presence of T. gondii antibodies in both the species, with a higher likelihood of seropositivity among heavier animals and females, respectively. For raccoon dogs, juveniles were more likely to be seropositive than adults at a given weight. Our results suggest that T. gondii infection is widespread in the regional raccoon and raccoon dog populations, indicating a high level of parasite circulation in the environment.
RESUMO
Toxoplasma gondii is an obligate intracellular protozoan that causes toxoplasmosis in warm-blooded animals. Although most infections in humans and animals are subclinical, an infection can nevertheless be fatal. One of the important characteristics in the epidemiology of this parasite is waterborne transmission. The American mink (Neogale vison), a mammal closely adapted to freshwater ecosystems, is a potential sentinel for T. gondii. We analysed meat juice from the heart of 194 wild minks collected between 2019 and 2022 in five study areas from Germany and Poland and tested for the presence of antibodies against T. gondii. The analysis was performed using a commercial enzyme-linked immunosorbent assay test (ELISA). Antibodies were detected in 45.36% (88/194, 95% confidence interval (CI): 38.39-52.41%) of the analysed animals. While the prevalence values ranged from 37.50% to 49.30%, there was no significant difference in seroprevalence between the study areas. Juveniles were less likely to carry T. gondii antibodies than adults (odds ratio: 0.216), whereas there was no significant difference in prevalence between the sexes (odds ratio: 0.933). The results of our study show that contact with T. gondii is widespread in minks, and the parasite is common in inland freshwater ecosystems in Germany and Poland. This indicates that watercourses play an important role in the spread of T. gondii oocysts.
RESUMO
BACKGROUND: In January 2021, a female 1-year-old Kunekune was presented at the University Clinic for Swine with severe reduction of the field of vision resulting in prolonged reaction time when targeting barriers, due to moderate to severe thickening of the skin around both orbits also affecting the eyelids. METHODS: Clinical examination revealed skin hyperplasia, nodular enlargement of the skin pores of the axillar and inguinal region. Ophthalmologists decided to remove parts of the thickened periocular skin, followed by histopathological examination. RESULTS: Once large amounts of demodectic mites were detected by histopathology, demodicosis could be diagnosed and treatment of the pig was started using sarolaner. Morphological and molecular analyses were performed. Histopathological and parasitological exams led to the aetiological diagnosis of demodicosis in the affected Kunekune pig. Severe skin lesions were revealed to be the consequence of an infestation with Demodex sp. Morphological analyses confirmed the involvement of D. phylloides. Molecular characterization indicated a Demodex species closely related to mites documented in wild boar - most probably D. phylloides for which no explicit sequences are available in GenBank yet. Treatment with sarolaner (2.6 mg/kg) resulted in a substantial regression of skin lesions, already detectable 1 month after first treatment. CONCLUSIONS: Demodicosis is a very rare disease in pigs that is most probably related to an impaired immune response to the mites. Demodectic mange should be included in the list of differential diagnoses in cases of periocular alterations of the skin of pigs.
Assuntos
Azetidinas , Pele , Compostos de Espiro , Suínos , Animais , Feminino , Instituições de Assistência Ambulatorial , Diagnóstico DiferencialRESUMO
African swine fever (ASF) is one of the most important viral diseases of domestic pigs and wild boar. Apart from endemic cycles in Africa, ASF is now continuously spreading in Europe and Asia. As ASF leads to severe but unspecific clinical signs and high lethality, early pathogen detection is of utmost importance. Recently, 'point-of-care' (POC) tests, especially immunochromatographic assays, have been intensively discussed for the use in remote areas but also in the context of on-farm epidemiological investigations and wild boar carcass screening. The later topic was the starting point for our present study. In detail, we evaluated the performance of the commercially available INGEZIM ASFV CROM Ag lateral flow assay (Eurofins Technologies Ingenasa) with selected high-quality reference blood samples, and with blood samples from wild boar carcasses collected under field conditions in Germany. While we observed a sensitivity of roughly 77% in freeze-thawed matrices of close to ideal quality, our approach to simulate field conditions in direct testing of blood samples from carcasses without any modification, resulted in a drastically reduced sensitivity of only 12.5% with the given sample set. Freeze thawing increased the sensitivity to roughly 44% which mirrored the overall sensitivity of 49% in the total data set of wild boar carcass samples. A diagnostic specificity of 100% was observed. In summary, the antigen LFA should not be regarded as a substitute for any OIE listed diagnostic method and has very limited use for carcass testing at the point of care. For optimized LFA antigen tests, the sensitivity with field samples must be significantly increased. An improved sensitivity is seen with freeze-thawed samples, which may indicate problems in the accessibility of ASFV antigen that could be overcome, to a certain extent, with assay modifications.
Assuntos
Vírus da Febre Suína Africana , Febre Suína Africana , Doenças dos Suínos , Febre Suína Africana/diagnóstico , Febre Suína Africana/epidemiologia , Animais , Europa (Continente) , Alemanha , Sus scrofa , SuínosRESUMO
African swine fever (ASF) is a major threat to pig production, and real-time PCR (qPCR) protocols are an integral part of ASF laboratory diagnosis. With the pandemic spread of ASF, commercial kits have risen on the market. In Germany, the kits have to go through an approval process and thus, general validation can be assumed. However, they have never been compared to each other. In this study, 12 commercial PCR kits were compared to an OIE-recommended method. Samples representing different matrices, genome loads, and genotypes were included in a panel that was tested under diagnostic conditions. The comparison included user-friendliness, internal controls, and the time required. All qPCRs were able to detect ASFV genome in different matrices across all genotypes and disease courses. With one exception, there were no significant differences when comparing the overall mean. The overall specificity was 100% (95% CI 87.66-100), and the sensitivity was between 95% and 100% (95% CI 91.11-100). As can be expected, variability concerned samples with low genome load. To conclude, all tests were fit for purpose. The test system can therefore be chosen based on compatibility and prioritization of the internal control system.
Assuntos
Vírus da Febre Suína Africana/genética , Vírus da Febre Suína Africana/isolamento & purificação , Febre Suína Africana/diagnóstico , Febre Suína Africana/virologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Vírus da Febre Suína Africana/classificação , Criação de Animais Domésticos/organização & administração , Animais , DNA Viral/genética , Genoma Viral , Genótipo , Alemanha , Kit de Reagentes para Diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/instrumentação , Sensibilidade e Especificidade , Suínos , Organização Mundial da SaúdeRESUMO
African swine fever (ASF) is among the most devastating viral diseases of pigs and wild boar worldwide. In recent years, the disease has spread alarmingly. Despite intensive research activities, a commercialized vaccine is still not available, and efficacious live attenuated vaccine candidates raise safety concerns. From a safety perspective, inactivated preparations would be most favourable. However, both historical and more recent trials with chemical inactivation did not show an appreciable protective effect. Under the assumption that the integrity of viral particles could enhance presentation of antigens, we used gamma irradiation for inactivation. To this means, gamma irradiated ASFV "Estonia 2014" was adjuvanted with either Polygen™ or Montanide™ ISA 201 VG, respectively. Subsequently, five weaner pigs per preparation were immunized twice with a three-week interval. Six weeks after the first immunization, all animals were challenged with the highly virulent ASFV strain "Armenia 2008". Although ASFV p72-specific IgG antibodies were detectable in all vaccinated animals prior challenge, no protection could be observed. All animals developed an acute lethal course of ASF and had to be euthanized at a moderate humane endpoint within six days. Indeed, the vaccinated pigs showed even higher clinical scores and a higher inner body temperature than the control group. However, significantly lower viral loads were detectable in spleen and liver of immunized animals at the time point of euthanasia. This phenomenon suggests an immune mediated disease enhancement that needs further investigation.
Assuntos
Febre Suína Africana , Vacinas Virais , Febre Suína Africana/prevenção & controle , Vírus da Febre Suína Africana , Animais , Raios gama , Imunogenicidade da Vacina , Suínos , Vacinação , Vacinas Atenuadas/imunologia , Proteínas Virais , Vacinas Virais/imunologiaRESUMO
African swine fever (ASF) is a viral disease that affects members of the Suidae family. The notifiable disease is considered a major threat to the pig industry, animal health, and food security worldwide. According to the European Food Safety Authority, ASF virus (ASFV) survival and transmission in feed and feed materials is a major research gap. Against this background, the objective of this study was to determine the survival of ASFV on spiked spray-dried porcine plasma (SDPP) when stored at two different temperatures. To this means, commercial SDPP granules were contaminated with high titers of ASFV in a worst-case external contamination scenario. Three samples per time point and temperature condition were subjected to blind passaging on macrophage cultures and subsequent haemadsorption test to determine residual infectivity. In addition, viral genome was detected by real-time PCR. The results indicate that heavily contaminated SDPP stored at 4°C remains infectious for at least 5 weeks. In contrast, spiked SDPP stored at room temperature displayed a distinct ASFV titer reduction after 1 week (>2.8 log levels) and complete inactivation after 2 weeks (>5.7 log levels). In conclusion, the residual risk of ASFV transmission through externally contaminated SDPP is low if SDPP is stored at room temperature (21 ± 2°C) for a period of at least 2 weeks before feeding.
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Vírus da Febre Suína Africana , Febre Suína Africana , Doenças dos Suínos , Animais , Genoma Viral , Plasma , SuínosRESUMO
African swine fever virus (ASFV) causes a hemorrhagic disease in pigs with high socio-economic consequences. To lower the impact of disease incursions, early detection is crucial. In the context of experimental animal trials, we evaluated diagnostic workflows for a high sample throughput in active surveillance, alternative sample matrices for passive surveillance, and lateral flow devices (LFD) for rapid testing. We could demonstrate that EDTA blood is significantly better suited for early ASFV detection than serum. Tissues recommended by the respective diagnostic manuals were in general comparable in their performance, with spleen samples giving best results. Superficial lymph nodes, ear punches, and different blood swabs were also evaluated as potential alternatives. In summary, all matrices yielded positive results at the peak of clinical signs and could be fit for purpose in passive surveillance. However, weaknesses were discovered for some matrices when it comes to the early phase of infection or recovery. The antigen LFD showed variable results with best performance in the clinical phase. The antibody LFD was quite comparable with ELISA systems. Concluding, alternative approaches are feasible but have to be embedded in control strategies selecting test methods and sample materials following a "fit-for-purpose" approach.
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African swine fever (ASF) has evolved from an exotic animal disease to a threat to global pig production. An important avenue for the wide-spread transmission of animal diseases is their dissemination through boar semen used for artificial insemination. In this context, we investigated the role of male reproductive organs in the transmission of ASF. Mature domestic boars and adolescent wild boars, inoculated with different ASF virus strains, were investigated by means of virological and pathological methods. Additionally, electron microscopy was employed to investigate in vitro inoculated sperm. The viral genome, antigens and the infectious virus could be found in all gonadal tissues and accessory sex glands. The viral antigen and viral mRNAs were mainly found in mononuclear cells of the respective tissues. However, some other cell types, including Leydig, endothelial and stromal cells, were also found positive. Using RNAScope, p72 mRNA could be found in scattered halo cells of the epididymal duct epithelium, which could point to the disruption of the barrier. No direct infection of spermatozoa was observed by immunohistochemistry, or electron microscopy. Taken together, our results strengthen the assumption that ASFV can be transmitted via boar semen. Future studies are needed to explore the excretion dynamics and transmission efficiency.
Assuntos
Vírus da Febre Suína Africana/isolamento & purificação , Febre Suína Africana/transmissão , Febre Suína Africana/virologia , Genitália Masculina/virologia , Febre Suína Africana/patologia , Vírus da Febre Suína Africana/genética , Vírus da Febre Suína Africana/fisiologia , Animais , Glândulas Bulbouretrais/patologia , Glândulas Bulbouretrais/virologia , DNA Viral/análise , Epididimo/patologia , Epididimo/virologia , Genitália Masculina/patologia , Leucócitos Mononucleares/virologia , Masculino , Próstata/patologia , Próstata/virologia , RNA Mensageiro/análise , RNA Viral/análise , Espermatozoides/ultraestrutura , Espermatozoides/virologia , Sus scrofa , Suínos , Testículo/patologia , Testículo/virologia , Replicação ViralRESUMO
Infection with African swine fever virus (ASFV) causes a highly lethal haemorrhagic disease in domestic and Eurasian wild pigs. Thus, it is a major threat to pig populations worldwide and a cause of substantial economic losses. Recently, less virulent ASFV strains emerged naturally, which showed higher experimental virulence in wild boar than in domestic pigs. The reason for this difference in disease progression and outcome is unclear but likely involves different immunological responses. Unfortunately, besides the importance of CD8α+ lymphocytes, little is known about the immune responses against ASFV in suids. Against this background, we used a multicolour flow cytometry platform to investigate the T-cell responses in wild boar and domestic pigs after infection with the moderately virulent ASFV strain 'Estonia2014' in two independent trials. CD4- /CD8α+ and CD4+ /CD8α+ αß T-cell frequencies increased in both subspecies in various tissues, but CD8α+ γδ T cells differentiated and responded in wild boar only. Proliferation in CD8α+ T cells was found 10 days post infectionem only. Frequencies of T-bet+ T cells increased in wild boar but not in domestic pigs. Of note, we found a considerable loss of perforin expression in cytotoxic T cells, 5 and 7 dpi. Both subspecies established a regulatory T-cell response 10 dpi. In domestic pigs, we show increasing levels of ICOS+ and CD8α+ invariant Natural Killer T cells. These disparities in T-cell responses might explain some of the differences in disease progression in wild boar and domestic pigs and should pave the way for future studies.
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Vírus da Febre Suína Africana , Febre Suína Africana , Doenças dos Suínos , Linfócitos T , Animais , Sus scrofa , Suínos , VirulênciaRESUMO
African swine fever (ASF) is one of the most important and devastating viral diseases in wild boar and domestic pigs worldwide. In the absence of vaccines or treatment options, early clinical detection is crucial and requires a sound knowledge of disease characteristics. To provide practitioners and state veterinarians with detailed information, the objective of the present study was to characterize the ASF virus (ASFV) isolate "Belgium 2018/1" in subadult and weaning domestic pigs. To this end, two animal trials were performed. Trial A included eight subadult domestic pigs and trial B five weaner pigs. In general, clinical signs and pathological lesions were in line with previous studies utilizing highly virulent ASF genotype II viruses. However, in trial A, four subadult domestic pigs survived and recovered, pointing to an age-dependent outcome. The long-term fate of these survivors remains under discussion and would need further investigation.
RESUMO
African swine fever (ASF) is an infectious disease of pigs and represents a massive threat to animal health and the pig industry worldwide. The ASF virus (ASFV) is efficiently transmitted via blood and meat from infected animals and can be highly stable in the environment. There is therefore great concern about the potential role of contaminated raw materials used for feed or bedding in the spread of ASFV. Especially crops and derived products originating from areas with ASF in wild boar and thus with high environmental ASFV contamination may be a risk for virus introduction into domestic pig herds. However, little is known about the stability of ASFV on contaminated crops and possible inactivation methods. In this study, we tested the effect of drying and heat treatment on the inactivation of ASFV on six different types of field crops, namely wheat, barley, rye, triticale, corn, and peas, contaminated with infectious blood. Samples were analysed for the presence of viral DNA and infectious virus after 2 hr drying at room temperature or after drying and 1 hr exposure to moderate heat at a specific temperature between 40°C and 75°C. ASFV genome was detected in all samples by real-time polymerase chain reaction (PCR), including samples that had been dried for 2 hr and incubated for 1 hr at 75°C. On the other hand, no infectious virus could be detected after 2 hr drying using virus isolation in porcine macrophages in combination with the detection of ASFV by the haemadsorption test (HAT). We therefore conclude that the risk of ASFV transmission via contaminated crops is most likely low, if they are incubated for at least 2 hr minimum at room temperature. Nonetheless, to minimize the risk of transmission as much as possible crops from ASF-affected zones should not be used for pig feed.
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Vírus da Febre Suína Africana/fisiologia , Febre Suína Africana/virologia , Produtos Agrícolas/virologia , Temperatura Alta , Doenças dos Suínos/virologia , Inativação de Vírus/efeitos da radiação , Febre Suína Africana/sangue , Vírus da Febre Suína Africana/genética , Vírus da Febre Suína Africana/isolamento & purificação , Animais , DNA Viral/genética , Genoma Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Sus scrofa/virologia , Suínos , Doenças dos Suínos/sangueRESUMO
Endemically infected European wild boar are considered a major reservoir of African swine fever virus in Europe. While high lethality was observed in the majority of field cases, strains of moderate virulence occurred in the Baltic States. One of these, "Estonia 2014", led to a higher number of clinically healthy, antibody-positive animals in the hunting bag of North-Eastern Estonia. Experimental characterization showed high virulence in wild boar but moderate virulence in domestic pigs. Putative pathogenic differences between wild boar and domestic pigs are unresolved and comparative pathological studies are limited. We here report on a kinetic experiment in both subspecies. Three animals each were euthanized at 4, 7, and 10 days post infection (dpi). Clinical data confirmed higher virulence in wild boar although macroscopy and viral genome load in blood and tissues were comparable in both subspecies. The percentage of viral antigen positive myeloid cells tested by flow cytometry did not differ significantly in most tissues. Only immunohistochemistry revealed consistently higher viral antigen loads in wild boar tissues in particular 7 dpi, whereas domestic pigs already eliminated the virus. The moderate virulence in domestic pigs could be explained by a more effective viral clearance.
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African swine fever (ASF) is one of the most important and complex viral diseases in domestic pigs and wild boar. Over the last decade, the disease has spread to several European and Asian countries and is now one of the major threats to profitable pig production worldwide. One of the more recently affected western countries is Belgium. To date, only wild boar are affected in a rather defined area in the Luxembourg region close to France, Luxembourg and Germany. While detailed sequence analyses were recently performed, biological characterization was still pending. Here, we report on the experimental inoculation of four sub-adult wild boar to further characterize the virus and its distribution in different tissues. After oronasal inoculation with the virus strain 'Belgium 2018/1', all animals developed an acute and severe disease course with typical pathomorphological and histopathological lesions. Organs and blood samples were positive in qPCR, haemadsorption test and antigen lateral flow devices (LFD). Virus and viral genome were also detected in genitals and accessory sex glands of two boars. There were no antibodies detectable in commercial antibody ELISAs, antibody LFDs and indirect immunoperoxidase tests. Thus, the genotype II ASF virus isolate 'Belgium 2018/1' showed a highly virulent phenotype in European wild boar similar to parental viruses like Armenia 2007 and other previously characterized ASFV strains. The study also provided a large set of well-characterized sample materials for test validation and assay harmonization.
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Vírus da Febre Suína Africana/patogenicidade , Febre Suína Africana/virologia , Sus scrofa/virologia , Doenças dos Suínos/virologia , Febre Suína Africana/patologia , Vírus da Febre Suína Africana/imunologia , Vírus da Febre Suína Africana/isolamento & purificação , Animais , Animais Selvagens/virologia , Anticorpos Antivirais/sangue , Bélgica , Ensaio de Imunoadsorção Enzimática/veterinária , Genoma Viral , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Suínos , Doenças dos Suínos/patologia , Virulência/fisiologiaRESUMO
Over the last decade, African swine fever (ASF) has changed from an exotic disease of Sub-Saharan Africa to a considerable and serious threat to pig industry in Central Europe and Asia. With the introduction of genotype II strains into the European Union in 2014, the disease has apparently found a fertile breeding ground in the abundant wild boar population. Upon infection with highly virulent ASF virus (ASFV), a haemorrhagic fever like illness with high lethality is seen in naïve domestic pigs and wild boar. Despite intensive research, virulence factors, host-virus interactions and pathogenesis are still far from being understood, and neither vaccines nor treatment exist. However, to better understand the disease, and to work towards a safe and efficacious vaccine, this information is needed. The presented review targets the knowledge gained over the last five years with regard to ASF pathogenesis in the broader sense but with a focus on the pandemic genotype II strains. In this way, it is designed as an update and supplement to existing review articles on the same topic.