• Journal of Veterinary Science
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• v.22 no.2
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• pp.28.1-28.6
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• 2021

An African swine fever (ASF) outbreak in wild boars was first reported on October 2, 2019, in South Korea. Since then, additional cases were reported in South Korea's border areas. We here report the identification of ASF virus (ASFV) DNAs from two out of eight environmental abiotic matter samples collected from areas where ASF-positive wild boar carcasses were found. Comparative genomic investigations suggested that the contaminating ASFV DNAs originated from the wild boar whose carcass had been found near the positive sample sites. This is the first report on the identification of ASF viral material in wild boar habitats.

• Journal of Veterinary Science
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• v.23 no.4
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• pp.51.1-51.10
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• 2022

Background: Due to the unavailability of an effective vaccine or antiviral drug against the African swine fever virus (ASFV), rapid diagnosis methods are needed to prevent highly contagious African swine fever. Objectives: The objective of this study was to establish the ladder-shape melting temperature isothermal amplification (LMTIA) assay for the detection of ASFV. Methods: LMTIA primers were designed with the p72 gene of ASFV as the target, and plasmid pUC57 was used to clone the gene. The LMTIA reaction system was optimized with the plasmid as the positive control, and the performance of the LMTIA assay was compared with that of the commercial real-time polymerase chain reaction (PCR) kit in terms of sensitivity and detection rate using 200 serum samples. Results: Our results showed that the LMTIA assay could detect the 10⁴ dilution of DNA extracted from the positive reference serum sample, which was the same as that of the commercial real-time PCR kit. The coincidence rate between the two assays was 100%. Conclusions: The LMTIA assay had high sensitivity, good detection, and simple operation. Thus, it is suitable for facilitating preliminary and cost-effective surveillance for the prevention and control of ASFV.

• Korean Journal of Veterinary Research
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• v.45 no.1
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• pp.63-70
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• 2005

African swine fever (ASF) is an infectious disease of domestic and wild pigs for which there is no vaccine in the world. A proper surveillance of viral activity and a timely response to ASF outbreaks depend upon the rapid diagnosis of ASF viral infection. Internationally prescribed enzyme-linked immunosorbent assay (ELISA) is a fast, sensitive test routinely used in the diagnosis of the ASF. However, inactivated whole ASF virus antigen used in this test is a tedious to prepare and has a risk of outside exposure of infectious virus by laboratory accident during the preparation. An ASF virus noninfectious recombinant antigen is a safe and easily produced alternative antigen for use in diagnostic assay. We have cloned the ORF O61R gene of the ASF virus to generate a recombinant baculovirus producing the p12 protein in insect cells under control of the polyhedrin promoter as non-fusion protein. When used in an indirect ELISA, the p12 antigen showed reactivity with all known ASF positive pig sera but not with negative pig sera. Our results indicated that the p12 protein would be one of alternative antigens for diagnosis of the ASF.

• Journal of Veterinary Science
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• v.22 no.6
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• pp.87.1-87.12
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• 2021

Background: African swine fever virus (ASFV), classical swine fever virus (CSFV), and porcine reproductive and respiratory syndrome virus (PRRSV) are still prevalent in many regions of China. Co-infections make it difficult to distinguish their clinical symptoms and pathological changes. Therefore, a rapid and specific method is needed for the differential detection of these pathogens. Objectives: The aim of this study was to develop a multiplex real-time quantitative reverse transcription polymerase chain reaction (multiplex qRT-PCR) for the simultaneous differential detection of ASFV, CSFV, and PRRSV. Methods: Three pairs of primers and TaqMan probes targeting the ASFV p72 gene, CSFV 5' untranslated region, and PRRSV ORF7 gene were designed. After optimizing the reaction conditions, including the annealing temperature, primer concentration, and probe concentration, multiplex qRT-PCR for simultaneous and differential detection of ASFV, CSFV, and PRRSV was developed. Subsequently, 1,143 clinical samples were detected to verify the practicality of the assay. Results: The multiplex qRT-PCR assay could specifically and simultaneously detect the ASFV, CSFV, and PRRSV with a detection limit of 1.78 × 10⁰ copies for the ASFV, CSFV, and PRRSV, but could not amplify the other major porcine viruses, such as pseudorabies virus, porcine circovirus type 1 (PCV1), PCV2, PCV3, foot-and-mouth disease virus, porcine parvovirus, atypical porcine pestivirus, and Senecavirus A. The assay had good repeatability with coefficients of variation of intra- and inter-assay of less than 1.2%. Finally, the assay was used to detect 1,143 clinical samples to evaluate its practicality in the field. The positive rates of ASFV, CSFV, and PRRSV were 25.63%, 9.36%, and 17.50%, respectively. The co-infection rates of ASFV+CSFV, ASFV+PRRSV, CSFV+PRRSV, and ASFV+CSFV+PRRSV were 2.45%, 2.36%, 1.57%, and 0.17%, respectively. Conclusions: The multiplex qRT-PCR developed in this study could provide a rapid, sensitive, specific diagnostic tool for the simultaneous and differential detection of ASFV, CSFV, and PRRSV.

• Journal of Ecology and Environment
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• v.46 no.1
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• pp.54-61
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• 2022

Background: In South Korea, African swine fever virus (ASFV) has spread among wild boars through Gangwon-do to Dangyang-gun, Chungcheongbuk-do on the southern border of Gangwon-do. To prevent the spread of ASFV to African swine fever (ASF)-free areas, it is necessary to identify areas with a high probability of finding ASFV-infected carcasses and to reduce the density of wild boars in those areas. In this study, we described the propagation trend of ASFV among wild boars, constructed the habitat suitability maps for ASFV-infected carcasses, and suggested areas with a high probability of finding ASFV-infected carcasses and an important route of ASFV transmission. Results: Despite the active quarantine policies in Korea to prevent the spread of ASFV through wild boars, there was no significant difference in the monthly average of number of ASFV-infected carcasses observed between 2020 and 2021. The ASFV-infected carcasses were found more in winter and spring (January to April). Since the first ASF outbreak in wild boars on October 2, 2019, the maximum width of ASFV-infected carcass distribution area was 222.7 km for about 26 months till November 20, 2021. The habitat suitability map, based on GPS coordinates of ASFV-infected wild boar carcasses, shows that highly detectable areas of ASFV-infected carcasses were sporadically dispersed in western and southwestern parts of Gangwon-do, and ranged from north to south of the province along the Baekdudaegan Mountains, whereas poorly detectable areas ranged along the north to the south in the middle parts of the province. Conclusions: Our suitability model, based on the GPS coordinates of ASFV-infected carcasses, identifies potential habitats where ASFV-infected carcasses are likely to be found and ponential routes where ASFV is likely to spread. Among ASF-free areas, the areas with high suitability predicted in this study should be given priority as survey areas to find ASFV-infected carcasses and hunting areas to reduce wild boar populations.

• Korean Journal of Veterinary Service
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• v.45 no.1
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• pp.13-18
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• 2022

African swine fever (ASF) is fatal to domestic pigs and wild boars (Sus scrofa) and affects the domestic pig industry. ASF is transmitted directly through the secretions of infected domestic pigs or wild boars, an essential source of infection in disease transmission. ASFV is also very stable in the environment. Thus, the virus is detected in the surrounding environment where ASF-infected carcasses are found. In this study, ASF infection monitoring was conducted on the swab and whole blood samples from wild animals, various hematopoietic arthropod samples that could access infected wild boar carcasses or habitats to cause maintenance and spread of disease, and soil samples of wild boar habitats. ASF viral DNA detection was confirmed negative in 317 wildlife and environmental samples through a real-time polymerase chain reaction. However, ASF occurs in the wild boars and spreads throughout the Korean peninsula. Therefore, it is necessary to trace the route of ASF virus infection by a continuous vector. Additional monitoring of various samples with potential ASF infection is needed to help the epidemiologic investigation and disease prevention.

• Journal of Veterinary Science
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• v.21 no.2
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• pp.38.1-38.24
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• 2020

African swine fever (ASF), caused by the ASF virus, a member of the Asfarviridae family, is one of the most important diseases in the swine industry due to its clinical and economic impacts. Since the first report of ASF a century ago, ample information has become available, but prevention and treatment measures are still inadequate. Two waves of epizootic outbreaks have occurred worldwide. While the first wave of the epizootic outbreak was controlled in most of the infected areas, the second wave is currently active in the European and Asian continents, causing severe economic losses to the pig industry. There are different patterns of spreading in the outbreaks between those in European and Asian countries. Prevention and control of ASF are very difficult due to the lack of available vaccines and effective therapeutic measures. However, recent outbreaks in South Korea have been successfully controlled on swine farms, although feral pigs are periodically being found to be positive for the ASF virus. Therefore, we would like to share our story regarding the preparation and application of control measures. The success in controlling ASF on farms in South Korea is largely due to the awareness and education of swine farmers and practitioners, the early detection of infected animals, the implementation of strict control policies by the government, and widespread sharing of information among stakeholders. Based on the experience gained from the outbreaks in South Korea, this review describes the current understanding of the ASF virus and its pathogenic mechanisms, epidemiology, and control.

• Journal of Veterinary Science
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• v.22 no.3
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• pp.35.1-35.13
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• 2021

Background: African swine fever (ASF) is an infectious viral disease of domestic pigs that presents as a hemorrhagic fever, and for which no effective vaccine is available. The disease has a serious negative social and economic impact on pig keepers. There is limited information on the potential risk factors responsible for the spread of ASF in South Kivu. Objective: The aim of this study was to determine the potential risk factors associated with ASF infection in suspected ASF virus (ASFV)-infected pigs. Methods: We sampled whole blood from 391 pigs. Additionally, 300 pig farmers were interviewed using a structured questionnaire. Viral DNA was detected by using the real-time polymerase chain reaction technique. Results: The majority of pigs sampled, 78% (95% confidence interval [CI], 74.4-82.6), were of local breeds. Over half, 60.4% (95% CI, 55.5-65.2), were female, and most of them, 90.5% (95% CI, 87.6-93.4), were adult pigs (> 1 year old). Viral DNA was detected in 72 of the 391 sampled pigs, indicating an overall infection rate of 18.4% (95% CI, 14.5-22.4). Multivariable logistic regression analysis revealed several risk factors positively associated with ASFV infection: feeding with swill in pen (odds ratio [OR], 3.8; 95% CI, 2.12-6.77); mixed ages of pigs in the same pen (OR, 3.3; 95% CI, 1.99-5.57); introduction of new animals to the farm (OR, 5.4; 95% CI, 1.91-15.28). The risk factors that were negatively (protective) correlated with ASFV positivity were the presence of male animals and the use of an in-pen breeding system. Conclusion: Local pig farmers should be encouraged to adopt proper husbandry and feeding practices in order to increase the number of ASF-free farms.

• Biomedical Science Letters
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• v.28 no.4
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• pp.334-339
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• 2022

African swine fever virus (ASFV) is a highly contagious and lethal pathogen that poses a threat to the global pork industry. The World Organization for Animal Health (WOAH) has placed strict surveillance measures for ASFV. The possibility of long-term survival of ASFV in raw meat or undercooked pork has been reported. Accordingly, the problem of secondary infection in food waste from households or waste disposal facilities has emerged, raising the need for ASFV monitoring of food waste. However, most of the previously reported ASFV gene detection methods are focused on clinical monitoring of pigs. There are very few cases in which their application in waste has been verified. Since ASFV diagnosis requires rapid monitoring and immediate action, loop-mediated isothermal amplification (LAMP) may be suitable, but this requires conformity assessment for LAMP to be used as a diagnostic technique. In this study, six LAMP methods were evaluated, and two methods (kit and manual) were recommended for use in diagnosing ASFV in food waste.

• Korean Journal of Veterinary Service
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• v.45 no.1
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• pp.19-30
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• 2022

In this study, a simple loop-mediated isothermal amplification (LAMP) combined with visual detection method (vLAMP) assay was developed for the rapid and specific detection of African swine fever virus (ASFV), overcoming the shortcomings of previously described LAMP assays that require additional detection steps or pose a cross-contamination risk. The assay results can be directly detected by the naked eye using hydroxynaphthol blue after incubation for 40 min at 62℃. The assay specifically amplified ASFV DNA and no other viral nucleic acids. The limit of detection of the assay was <50 DNA copies/reaction, which was ten times more sensitive than conventional polymerase chain reaction (cPCR) and comparable to real-time PCR (qPCR). For clinical evaluation, the ASFV detection rate of vLAMP was higher than cPCR and comparable to OIE-recommended qPCR, showing 100% concordance, with a κ value (95% confidence interval) of 1 (1.00~1.00). Considering the advantages of high sensitivity and specificity, no possibility for cross-contamination, and being able to be used as low-cost equipment, the developed vLAMP assay will be a valuable tool for detecting ASFV from clinical samples, even in resource-limited laboratories.