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Genetic characterisation of African swine fever virus in outbreaks in Ha Nam province, Red River Delta Region of Vietnam, and activity of antimicrobial products against virus infection in contaminated feed
2020
Tran, Ha Thi Thanh | Truong, Anh Duc | Ly, Duc Viet | Vũ, Thị Hảo | Hoàng, Văn Tuấn | Nguyễn, Thị Chính | Chu, Thi Nhu | Nguyen, Thi Huyen | Pham, Ngoc Thi | Nguyen, Tinh | Yersin, Andrew G. | Dang, Hoang Vu
African swine fever (ASF) was officially reported in Vietnam in February 2019 and spread across the whole country, affecting all 63 provinces and cities. In this study, ASF virus (ASFV) VN/Pig/HaNam/2019 (VN/Pig/HN/19) strain was isolated in primary porcine alveolar macrophage (PAM) cells from a sample originating from an outbreak farm in Vietnam’s Red River Delta region. The isolate was characterised using the haemadsorption (HAD) test, real-time PCR, and sequencing. The activity of antimicrobial feed products was evaluated via a contaminated ASFV feed assay. Phylogenetic analysis of the viral p72 and EP402R genes placed VN/Pig/HN/19 in genotype II and serogroup 8 and related it closely to Eastern European and Chinese strains. Infectious titres of the virus propagated in primary PAMs were 10⁶ HAD₅₀/ml. Our study reports the activity against ASFV VN/Pig/HN/19 strain of antimicrobial Sal CURB RM E Liquid, F2 Dry and K2 Liquid. Our feed assay findings suggest that the antimicrobial RM E Liquid has a strong effect against ASFV replication. These results suggest that among the Sal CURB products, the antimicrobial RM E Liquid may have the most potential as a mitigant feed additive for ASFV infection. Therefore, further studies on the use of antimicrobial Sal CURB RM E Liquid in vivo are required. Our study demonstrates the threat of ASFV and emphasises the need to control and eradicate it in Vietnam by multiple measures.
اظهر المزيد [+] اقل [-]African swine fever virus – persistence in different environmental conditions and the possibility of its indirect transmission
2019
Mazur-Panasiuk, Natalia | Żmudzki, Jacek | Woźniakowski, Grzegorz
Since 2007, African swine fever (ASF) has posed a serious threat to the European swine industry. In Poland, the numbers of reported outbreaks in pigs and affected areas grow every year. In 2018, the disease was noted in Western Europe, in Belgium specifically, where several hundred infected wild boars have been detected so far. In 2018, the virus unexpectedly emerged in pig holdings in eastern China, northern Mongolia, Vietnam, and Cambodia, causing worldwide concern about its further spread. Since there is still no vaccine available, the only approach to control the disease is biosecurity. Identification of potential sources of the virus is extremely important in light of its phenomenal survivability. The review summarises the current knowledge about ASFV survivability and resistance to environmental conditions, and discusses the role of indirect contact in spreading the disease.
اظهر المزيد [+] اقل [-]Selected aspects related to epidemiology, pathogenesis, immunity, and control of African swine fever
2016
Woźniakowski, Grzegorz | Frączyk, Magdalena | Niemczuk, Krzysztof | Pejsak, Zygmunt
African swine fever (ASF) is currently one of the most severe viral infections of domestic pigs, wild boars, and other hosts belonging to Suidae family. ASF is also considered as the most complex and devastating infectious and haemorrhagic disease of swine due to its severe socio-economic impact and transboundary character. ASF it is a notifiable disease and due to the lack of specific treatment and vaccine, the disease can be only limited by the administrative measures comprising wild boar hunting and stamping out of affected pigs. ASF occurred for the first time in Kenya in 1921 while in Europe (Portugal) the virus was detected at the end of the 1950s. In spite of successful eradication of this threat in a number of affected regions, the virus remains endemic in both feral and domestic pigs in Africa and Sardinia. The ‘new era’ of ASF started in 2007 after its re-introduction to Georgia. Following its intensive expansion, the virus spread to other Caucasian countries, including the territory of the Russian Federation. In 2014 the virus reached Ukraine, Belarus, and, consequently, European Union countries: Lithuania, Latvia, Estonia, and Poland. The occurrence of ASF in wild boars and pigs had a severe impact on both epidemiology and economy because of the national and international transport and trade consequences. Up to date, starting from the February 2014, eighty ASF cases in wild boar and three outbreaks in domestic pigs have been diagnosed. Taking into account the diverse rate of spread in Poland, this review aims to present and discuss the current state of knowledge on ASF including its epidemiology, pathology, transmission, and perspectives of control.
اظهر المزيد [+] اقل [-]Characteristics of selected active substances used in disinfectants and their virucidal activity against ASFV
2019
Juszkiewicz, Małgorzata | Walczak, Marek | Woźniakowski, Grzegorz
African swine fever (ASF), caused by African swine fever virus (ASFV), is currently one of the most important and serious diseases of pigs, mainly due to the enormous sanitary and socio-economic consequences. It leads to serious economic losses, not only because of the near 100% mortality rate, but also through the prohibitions of pork exports it triggers. Currently neither vaccines nor safe and effective chemotherapeutic agents are available against ASFV. The disease is controlled by culling infected pigs and maintaining high biosecurity standards, which principally relies on disinfection. Some countries have approved and/or authorised a list of biocides effective against this virus. This article is focused on the characteristics of chemical substances present in the most popular disinfectants of potential use against ASFV. Despite some of them being approved and tested, it seems necessary to perform tests directly on ASFV to ensure maximum effectiveness of the disinfectants in preventing the spread of ASF in the future.
اظهر المزيد [+] اقل [-]ASF -survivors’ sera do not inhibit African swine fever virus replication in vitro
2022
Walczak, Marek | Juszkiewicz, Małgorzata | Szymankiewicz, Krzesimir | Szczotka-Bochniarz, Anna | Woźniakowski, Grzegorz
African swine fever virus (ASFV) causes one of the most dangerous diseases of pigs and wild boar – African swine fever (ASF). Since its second introduction into Europe (in 2007), the disease has been spreading consistently, and now ASF-free European countries are at risk. Complex interactions between the host’s immune system and the virus have long prevented the development of a safe vaccine against ASF. This study analysed the possibility of neutralisation of the ASFV in vitro by sera collected from ASF-survivor animals. Two pig and three wild boar serum samples were collected from previously selected potential ASF survivors. All sera presented high antibody titres (>5 log₁₀/mL). Primary alveolar macrophages were cultured in growth medium containing 10% and 20% concentrations of selected sera and infected with a haemadsorbing ASFV strain (Pol18_28298_O111, genotype II). The progress of infection was investigated under a light microscope by observing the cytopathic effect (CPE) and the haemadsorption phenomenon. Growth kinetics were investigated using a real-time PCR assay. Haemadsorption inhibition was detected in the presence of almost all selected sera; however, the inhibition of virus replication in vitro was excluded. In all samples, a CPE and decreasing quantification cycle values of the viral DNA were found. Anti-ASFV antibodies alone are not able to inhibit virus replication. Interactions between the humoral and cellular immune response which effectively combat the disease are implicated in an ASF-survivor’s organism.
اظهر المزيد [+] اقل [-]African swine fever (ASF) and ticks. No risk of tick-mediated ASF spread in Poland and Baltic states
2017
Frant, Maciej | Woźniakowski, Grzegorz | Pejsak, Zygmunt
Infectious diseases of swine, particularly zoonoses, have had a significant influence on nutritional safety and availability of pig meat as high-energy protein product since the time that pigs were domesticated back in the 7ᵗʰ century BC. The main sources of swine infectious diseases include the so-called primary sources (direct infection, i.e. through contact with infected and sick animals) and secondary sources (contaminated meat products, slaughter products, and vectors, including ticks). At present, the most serious epidemiological and economic threat to swine breeding in Europe is African swine fever (ASF). This disease, originally coming from Africa, is incurable and causes death of infected pigs and wild boars during 7−10 days after infection. Among the various factors that influence the spread of ASF, important role is played by ticks from the genus Ornithodoros, mainly from the species Ornithodoros moubata. Research on the ASF indicates that other species of ticks can also transmit the virus to healthy pigs in laboratory conditions. Sylvatic and domestic cycles of ASF virus transmission, which have been described so far, require further studies and updating in order to point the potential new vectors in the Caucasus and Eastern Europe affected by the ASF. Effective methods of control and biosecurity may significantly slow down the spread of ASF, which undoubtedly is a major threat to world pig production and international swine trade.
اظهر المزيد [+] اقل [-]Roles of African swine fever virus structural proteins in viral infection
2017
Jia, Ning | Ou, Yunwen | Pejsak, Zygmunt | Zhang, Yongguang | Zhang, Jie
African swine fever virus (ASFV) is a large, double-stranded DNA virus and the sole member of the Asfarviridae family. ASFV infects domestic pigs, wild boars, warthogs, and bush pigs, as well as soft ticks (Ornithodoros erraticus), which likely act as a vector. The major target is swine monocyte-macrophage cells. The virus can cause high fever, haemorrhagic lesions, cyanosis, anorexia, and even fatalities in domestic pigs. Currently, there is no vaccine and effective disease control strategies against its spread are culling infected pigs and maintaining high biosecurity standards. African swine fever (ASF) spread to Europe from Africa in the middle of the 20ᵗʰ century, and later also to South America and the Caribbean. Since then, ASF has spread more widely and thus is still a great challenge for swine breeding. The genome of ASFV ranges in length from about 170 to 193 kbp depending on the isolate and contains between 150 and 167 open reading frames (ORFs). The ASFV genome encodes 150 to 200 proteins, around 50 of them structural. The roles of virus structural proteins in viral infection have been described. These proteins, such as pp220, pp62, p72, p54, p30, and CD2v, serve as the major component of virus particles and have roles in attachment, entry, and replication. All studies on ASFV proteins lay a good foundation upon which to clarify the infection mechanism and develop vaccines and diagnosis methods. In this paper, the roles of ASFV structural proteins in viral infection are reviewed.
اظهر المزيد [+] اقل [-]A recombinase polymerase amplification-based assay for rapid detection of African swine fever virus
2017
Wang, Jianchang | Wang, Jinfeng | Geng, Yunyun | Yuan, Wanzhe
A recombinase polymerase amplification (RPA)-based method was developed for rapid and specific detection of African swine fever virus (ASFV), the etiologic agent of African swine fever, a devastating disease of swine. Primers and the exo probe targeting the conserved region of the P72 gene of ASFV were designed and the reaction was run on the Genie III scanner device. Using recombinant plasmid DNA containing the P72 gene as template, we showed that the amplified product could be detected in less than 10 min and that the detection limit was 102 copies DNA/reaction [same detection limit as real-time polymerase chain reaction (PCR)]. The RPA assay did not cross-detect CSFV, PCV-2, PRV, PRRSV, or FMDV, common viruses seen in pigs. Tests of recombinant plasmid-spiked serum samples revealed that RPA and real-time PCR had the same diagnostic rate. The RPA assay, which is simple, cost-effective, and fast, is a promising alternative to real-time PCR for ASFV detection.
اظهر المزيد [+] اقل [-]Detection of African swine fever viral antigens in paraffin-embedded tissues by use of immunohistologic methods and polyclonal antibodies
1992
Martínez Fernández, Ángel | Pérez, J. | Carrasco, L. | Sierra, M.A. | Sanchez-Vizcaino, M. | Jover, A.
Tissues obtained from pigs inoculated with African swine fever virus (ASFV), fixed by vascular perfusion using glutaraldehyde, and embedded in paraffin or araldite were used for an immunohistologic electron microscopic study. To detect ASFV antigens, 4 methods were used on paraffin sections with or without pretreatment of the tissues. Use of biotinylated anti-ASFV antiserum combined with avidin -biotin complex and peroxidase proved to be the most suitable method, and antigen was detected in tissues infected with 2 ASF viruses of different virulence. Use of the glutaraldehyde fixation method should ensure optimal morphologic (structural and ultrastructural) data while allowing an immunohistologic study, and add to knowledge of the pathogenesis of ASF.
اظهر المزيد [+] اقل [-]Comparison of two antigens for use in an enzyme-linked immunosorbent assay to detect African swine fever antibody
1990
Pastor, M.J. | Arias, M. | Escribano, J.M.
Two African swine fever virus (ASFV) antigens were tested for use in an ELISA to detect antibody to ASFV. Antigens used were the cytoplasmic soluble fraction (CS-P) of infected cells grown in the presence of porcine serum and the semipurified viral structural protein VP73 (SVP73). Both antigens were tested by ELISA against 72 sera obtained during several ASF field episodes and from ASFV-inapparent carriers. Of the 72 sera, only 2.8% had positive results by ELISA against CS-P antigen; 60% of positive-reacting sera (to both antigens) had higher ELISA values when the CS-P antigen was used. Samples (with positive results) that reacted only to CS-P antigen had results confirmed by immunoblot analysis. Such sera reacted against ASFV-infection proteins IP25, IP25.5, and IP30, but not against IP73. In time-course experiments to detect appearance of ASFV-antibodies in infected miniature pigs, antibodies were detected by immunoblot analysis on postinoculation day (PID) 8. At that time, only the polypeptides IP25, IP25.5, IP30, and IP31 were recognized; IP73 and IP12 were first detected 3 and 4 days later, respectively. In the same experiments, ASFV antibodies were detected by ELISA, using CS-P or SVP73 antigens, on PID 7 and 9, respectively. These results could explain the percentage of sera not having positive results by ELISA using SVP73 antigen, if the sera were obtained from ASFV-infected pigs during the first days of infection before induction of antibody response against the IP73 protein. This feature makes the use of CS-P antigen advantageous in early serologic detection of AFSV-infected pigs.
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