Effectiveness of Chemical Compounds Used against African Swine Fever Virus in Commercial Available Disinfectants

African swine fever (ASF) causes huge economic losses and is one of most dangerous diseases of pigs. The disease is known for almost 100 years, an effective vaccine or treatment is still unavailable, only proper biosecurity measures, including disinfection, are being applied, in order to prevent disease outbreaks. Eight active substances, i.e., formaldehyde, sodium hypochlorite, caustic soda, glutaraldehyde, phenol, benzalkonium chloride, potassium peroxymonosulfate and acetic acid, were tested, in order to confirm their effectiveness against African swine fever virus (ASFV). This specific selection was done based on the World Organisation for Animal Health (OIE)’s recommendation and previous disinfectant studies on surfaces. The result of our study shows that most of them inactivate the virus, in recommended concentrations. In order to reduce the cytotoxicity of the four substances, Microspin S-400 HR columns were applied, therefore making it possible to demonstrate four logarithms virus titer reduction. Sodium hypochlorite, glutaraldehyde, caustic soda and potassium peroxymonosulfate showed the best ASFV inactivation rates, achieving titer reductions over 5 logs. Despite microfiltration, the virucidal activity of formaldehyde was not assessable, due to its high cytotoxicity. Our results showed that cleaning is particularly important, because removal of the soiling provides improved effectiveness of the tested chemical compounds.

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Structure of African Swine Fever Virus and Associated Molecular Mechanisms Underlying Infection and Immunosuppression: A Review

Frontiers in Immunology ◽

10.3389/fimmu.2021.715582 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yue Wang ◽

Weifang Kang ◽

Wenping Yang ◽

Jing Zhang ◽

Dan Li ◽

...

Keyword(s):

Immune Response ◽

Molecular Mechanism ◽

Molecular Mechanisms ◽

African Swine Fever Virus ◽

Animal Health ◽

African Swine Fever ◽

Fever Virus ◽

Economic Losses ◽

Cell Immune Response ◽

Effective Vaccine

African swine fever (ASF) is an acute, highly contagious, and deadly infectious disease. The mortality rate of the most acute and acute ASF infection is almost 100%. The World Organization for Animal Health [Office International des épizooties (OIE)] lists it as a legally reported animal disease and China lists it as class I animal epidemic. Since the first diagnosed ASF case in China on August 3, 2018, it has caused huge economic losses to animal husbandry. ASF is caused by the African swine fever virus (ASFV), which is the only member of Asfarviridae family. ASFV is and the only insect-borne DNA virus belonging to the Nucleocytoplasmic Large DNA Viruses (NCLDV) family with an icosahedral structure and an envelope. Till date, there are still no effective vaccines or antiviral drugs for the prevention or treatment of ASF. The complex viral genome and its sophisticated ability to regulate the host immune response may be the reason for the difficulty in developing an effective vaccine. This review summarizes the recent findings on ASFV structure, the molecular mechanism of ASFV infection and immunosuppression, and ASFV-encoded proteins to provide comprehensive proteomic information for basic research on ASFV. In addition, it also analyzes the results of previous studies and speculations on the molecular mechanism of ASFV infection, which aids the study of the mechanism of clinical pathological phenomena, and provides a possible direction for an intensive study of ASFV infection mechanism. By summarizing the findings on molecular mechanism of ASFV- regulated host cell immune response, this review provides orientations and ideas for fundamental research on ASFV and provides a theoretical basis for the development of protective vaccines against ASFV.

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Computational Analysis of African Swine Fever Virus Protein Space for the Design of an Epitope-Based Vaccine Ensemble

Pathogens ◽

10.3390/pathogens9121078 ◽

2020 ◽

Vol 9 (12) ◽

pp. 1078 ◽

Cited By ~ 1

Author(s):

Albert Ros-Lucas ◽

Florencia Correa-Fiz ◽

Laia Bosch-Camós ◽

Fernando Rodriguez ◽

Julio Alonso-Padilla

Keyword(s):

T Cell ◽

Vaccine Development ◽

De Novo ◽

African Swine Fever Virus ◽

African Swine Fever ◽

Fever Virus ◽

Economic Losses ◽

Virus Protein ◽

Hemorrhagic Disease ◽

Starting Point

African swine fever virus is the etiological agent of African swine fever, a transmissible severe hemorrhagic disease that affects pigs, causing massive economic losses. There is neither a treatment nor a vaccine available, and the only method to control its spread is by extensive culling of pigs. So far, classical vaccine development approaches have not yielded sufficiently good results in terms of concomitant safety and efficacy. Nowadays, thanks to advances in genomic and proteomic techniques, a reverse vaccinology strategy can be explored to design alternative vaccine formulations. In this study, ASFV protein sequences were analyzed using an in-house pipeline based on publicly available immunoinformatic tools to identify epitopes of interest for a prospective vaccine ensemble. These included experimentally validated sequences from the Immune Epitope Database, as well as de novo predicted sequences. Experimentally validated and predicted epitopes were prioritized following a series of criteria that included evolutionary conservation, presence in the virulent and currently circulating variant Georgia 2007/1, and lack of identity to either the pig proteome or putative proteins from pig gut microbiota. Following this strategy, 29 B-cell, 14 CD4+ T-cell and 6 CD8+ T-cell epitopes were selected, which represent a starting point to investigating the protective capacity of ASFV epitope-based vaccines.

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Development of a Directly Visualized Recombinase Polymerase Amplification–SYBR Green I Method for the Rapid Detection of African Swine Fever Virus

Frontiers in Microbiology ◽

10.3389/fmicb.2020.602709 ◽

2020 ◽

Vol 11 ◽

Author(s):

Shuai Zhang ◽

Aijun Sun ◽

Bo Wan ◽

Yongkun Du ◽

Yanan Wu ◽

...

Keyword(s):

African Swine Fever Virus ◽

African Swine Fever ◽

Sybr Green ◽

Fever Virus ◽

Diagnostic Methods ◽

Sybr Green I ◽

Recombinase Polymerase Amplification ◽

Economic Losses ◽

Field Samples ◽

Standard Plasmid

African swine fever (ASF) is a lethal disease in swine caused by etiologic African swine fever virus (ASFV). The global spread of ASFV has resulted in huge economic losses globally. In the absence of effective vaccines or drugs, pathogen surveillance has been the most important first-line intervention to prevent ASF outbreaks. Among numerous diagnostic methods, recombinase polymerase amplification (RPA)-based detection is capable of producing sensitive and specific results without relying on the use of expensive instruments. However, currently used gene-specific, probe-based RPA for ASFV detection is expensive and time-consuming. To improve the efficiency of ASFV surveillance, a novel directly visualized SYBR Green I-staining RPA (RPAS) method was developed to detect the ASFV genome. SYBR Green I was added to the amplified RPA products for direct visualization by the naked eye. The sensitivity and specificity of this method were confirmed using standard plasmid and inactivated field samples. This method was shown to be highly specific with a detection limit of 103 copies/μl of ASFV in 15 min at 35°C without any cross-reactions with other important porcine viruses selected. In summary, this method enables direct sample visualization with reproducible results for ASFV detection and hence has the potential to be used as a robust tool for ASF prevention and control.

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A Simple Method for Sample Preparation to Facilitate Efficient Whole-Genome Sequencing of African Swine Fever Virus

Viruses ◽

10.3390/v11121129 ◽

2019 ◽

Vol 11 (12) ◽

pp. 1129 ◽

Cited By ~ 9

Author(s):

Ferenc Olasz ◽

István Mészáros ◽

Szilvia Marton ◽

Győző L. Kaján ◽

Vivien Tamás ◽

...

Keyword(s):

Sample Preparation ◽

African Swine Fever Virus ◽

Animal Health ◽

African Swine Fever ◽

Fever Virus ◽

Whole Genome ◽

Swine Industry ◽

Viral Dna ◽

Simple Method ◽

Generation Sequencing

In the recent years, African swine fever has become the biggest animal health threat to the swine industry. To facilitate quick genetic analysis of its causative agent, the African swine fever virus (ASFV), we developed a simple and efficient method for next generation sequencing of the viral DNA. Execution of the protocol does not demand complicated virus purification steps, enrichment of the virus by ultracentrifugation or of the viral DNA by ASFV-specific PCRs, and minimizes the use of Sanger sequencing. Efficient DNA-se treatment, monitoring of sample preparation by qPCR, and whole genome amplification are the key elements of the method. Through detailed description of sequencing of the first Hungarian ASFV isolate (ASFV_HU_2018), we specify the sensitive steps and supply key reference numbers to assist reproducibility and to facilitate the successful use of the method for other ASFV researchers.

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JOURNEY OF AFRICAN SWINE FEVER VIRUS IN EASTERN EU STATES

CBU International Conference Proceedings ◽

10.12955/cbup.v6.1262 ◽

2018 ◽

Vol 6 ◽

pp. 863-869

Author(s):

Stelian Baraitareanu ◽

Dragos Cobzariu ◽

Mihaela Popp ◽

Marius Valer Campeanu ◽

Doina Danes

Keyword(s):

Czech Republic ◽

African Swine Fever Virus ◽

Animal Health ◽

African Swine Fever ◽

Fever Virus ◽

The Black Sea ◽

Eastern European ◽

The European Union ◽

The Czech Republic ◽

Backyard Pigs

INTRODUCTION: In 2007, African swine fever virus (ASFv) broken its well-known boundaries. This was the reference year for the first report of African swine fever (ASF) in Georgia. Subsequently, the virus reached pigs and boars in Armenia and Russia. From the Caucasus area, ASFv jumped in all directions, between the Black Sea and the Caspian Sea, in relation to the density of backyard pigs and their trade. In the next ten years there have been notifications and registrations of ASFv outbreaks in Russia, Azerbaijan, Ukraine, Belarus, Lithuania, Poland, Estonia, Latvia, Moldova and the Czech Republic. Romania faced the first ASFv outbreak at the end of July 2017, in backyard pigs." in stead "density of backyard pigs and their trade. In the next ten years there have been notifications and registrations of ASFv outbreaks in Russia, Azerbaijan, Ukraine, Belarus, Lithuania, Poland, Estonia, Latvia, Moldova and the Czech Republic. Romania faced the first ASFv outbreak at the end of July 2017, in backyard pigs.OBJECTIVES: The aim of study is to analyse the ways ASFv spread from and into different regions recorded by Eastern European states.METHODS: The immediate notifications on ASFv to the World Organisation for Animal Health (OIE) were analysed from the Eastern-European states between 2007 and 2017. The analysis took into consideration the first occurrence of the disease under scrutiny in the country and the follow-up reports, in relation with the geospatial distribution of the outbreaks.RESULTS: The main route of ASFv introduction into local pig populations indicated by the Member States of the European Union was the trans-boundary circulation of boars. However, the spread of ASFv through both, wild and domestic pigs and also by the human alimentary customs/traditions in the affected areas shouldn’t be ignored. Three cycles of ASFv transmission have been identified and described by the epidemiologists: the domestic cycle, the sylvatic cycle and the tick-pig cycle.CONCLUSION: None of the ways to disseminate the ASFv should be excluded, and the origin of the first outbreaks remains unknown or inconclusive in Eastern EU states.

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An Isothermal Molecular Point of Care Testing for African Swine Fever Virus Using Recombinase-Aided Amplification and Lateral Flow Assay Without the Need to Extract Nucleic Acids in Blood

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.633763 ◽

2021 ◽

Vol 11 ◽

Author(s):

Yuhang Zhang ◽

Qingmei Li ◽

Junqing Guo ◽

Dongliang Li ◽

Li Wang ◽

...

Keyword(s):

African Swine Fever Virus ◽

Point Of Care ◽

African Swine Fever ◽

Lateral Flow ◽

Fever Virus ◽

Lateral Flow Assay ◽

Economic Losses ◽

Point Of Care Testing ◽

Viral Dna ◽

Blood Samples

African swine fever (ASF) is a highly contagious and usually deadly porcine infectious disease listed as a notifiable disease by the World Organization for Animal Health (OIE). It has brought huge economic losses worldwide, especially since 2018, the first outbreak in China. As there are still no effective vaccines available to date, diagnosis of ASF is essential for its surveillance and control, especially in areas far from city with limited resources and poor settings. In this study, a sensitive, specific, rapid, and simple molecular point of care testing for African swine fever virus (ASFV) B646L gene in blood samples was established, including treatment of blood samples with simple dilution and boiling for 5 min, isothermal amplification with recombinase-aided amplification (RAA) at 37°C in a water bath for 10 min, and visual readout with lateral flow assay (LFA) at room temperature for 10–15 min. Without the need to extract viral DNA in blood samples, the intact workflow from sampling to final diagnostic decision can be completed with minimal equipment requirement in 30 min. The detection limit of RAA-LFA for synthesized B646L gene-containing plasmid was 10 copies/μl, which was 10-fold more sensitive than OIE-recommended PCR and quantitative PCR. In addition, no positive readout of RAA-LFA was observed in testing classical swine fever virus, porcine reproductive and respiratory syndrome virus, porcine epidemic diarrhea virus, pseudorabies virus and porcine circovirus 2, exhibiting good specificity. Evaluation of clinical blood samples of RAA-LFA showed 100% coincident rate with OIE-recommended PCR, in testing both extracted DNAs and treated bloods. We also found that some components in blood samples greatly inhibited PCR performance, but had little effect on RAA. Inhibitory effect can be eliminated when blood was diluted at least 32–64-fold for direct PCR, while only a 2–4 fold dilution of blood was suitable for direct RAA, indicating RAA is a better choice than PCR when blood is used as detecting sample. Taken together, we established an sensitive, specific, rapid, and simple RAA-LFA for ASFV molecular detection without the need to extract viral DNA, providing a good choice for point of care testing of ASF diagnosis in the future.

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DEVELOPMENT OF RECOMBINANT POSITIVE CONTROL FOR AFRICAN SWINE FEVER VIRUS PCR DETECTION

Biotechnologia Acta ◽

10.15407/biotech13.06.058 ◽

2020 ◽

Vol 13 (6) ◽

pp. 58-63

Author(s):

M. Kit ◽

Keyword(s):

African Swine Fever Virus ◽

Animal Health ◽

African Swine Fever ◽

Positive Control ◽

Fever Virus ◽

Laboratory Diagnostics ◽

E Coli ◽

Pcr Assays ◽

World Organisation ◽

Positive Controls

Recombinant plasmids containing target sequences are widely used as positive controls for PCR laboratory diagnostics. The aim of the study was development of recombinant positive control containing a fragment of B646L gene of African swine fever virus. The sequence of interest encodes targets of all the PCR assays for African swine fever laboratory diagnostics recommended by World Organisation for Animal Health. A plasmid containing 1763 bp insertion was cloned in E .coli DH5α strain. After purification, the plasmid ten-fold serial dulutions were used as a positive control while PRC testing. A minimal detectable copy number was 20 copies per reaction for both conventional and real-time PCR assays. The developed plasmid could be used as a safe and effective positive control while ASF laboratory diagnostics by PCR.

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Deletion of A137R gene from the pandemic strain of African swine fever virus is attenuated and offers protection against virulent pandemic virus

Journal of Virology ◽

10.1128/jvi.01139-21 ◽

2021 ◽

Author(s):

Douglas P. Gladue ◽

Elizabeth Ramirez-Medina ◽

Elizabeth Vuono ◽

Ediane Silva ◽

Ayushi Rai ◽

...

Keyword(s):

East Asia ◽

Central Europe ◽

Vaccine Candidate ◽

African Swine Fever Virus ◽

African Swine Fever ◽

Fever Virus ◽

Economic Losses ◽

Live Attenuated Vaccine ◽

Attenuated Vaccine ◽

Highly Virulent

African swine fever virus (ASFV) is causing a devastating pandemic in domestic and wild swine within an extended geographical area from Central Europe to East Asia resulting in economic losses for the regional swine industry. There are no commercial vaccines, therefore disease control relies on identification and culling of infected animals. We report here that the deletion of the ASFV gene A137R from the highly virulent ASFV-Georgia2010 (ASFV-G) isolate induces a significant attenuation of virus virulence in swine. A recombinant virus lacking the A137R gene, ASFV-G-ΔA137R, was developed to assess the role of this gene in ASFV virulence in domestic swine. Animals inoculated intramuscularly with 10 2 HAD 50 of ASFV-G-ΔA137R remained clinically healthy during the 28 day observational period. All animals inoculated with ASFV-G-ΔA137R had medium to high viremia titers and developed a strong virus-specific antibody response. Importantly, all ASFV-G-ΔA137R-inoculated animals were protected when challenged with the virulent parental strain ASFV-G. No evidence of replication of challenge virus was observed in the ASFV-G-ΔA137R-inoculated animals. Therefore, ASFV-G-ΔA137R is a novel potential live attenuated vaccine candidate and one of the few experimental vaccine strains reported to induce protection against the highly virulent ASFV Georgia virus that is the cause of the current Eurasian pandemic. Importance: No commercial vaccine is available to prevent African swine fever. The ASF pandemic caused by ASFV Georgia2007 (ASFV-G) is seriously affecting pork production in a contiguous area from Central Europe to East Asia. Here we report the rational development of a potential live attenuated vaccine strain by deleting a virus-specific gene, A137R, from the genome of ASFV-G. The resulting virus presented a completely attenuated phenotype and, importantly, animals infected with this genetically modified virus were protected from developing ASF after challenge with the virulent parental virus. ASFV-G-ΔA137R confers protection even at low doses (10 2 HAD 50 ) demonstrating its potential as a vaccine candidate. Therefore, ASFV-G-ΔA137R is a novel experimental ASF vaccine protecting pigs from the epidemiologically relevant ASFV Georgia isolate.

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African Swine Fever in Cameroon: A Review

Pathogens ◽

10.3390/pathogens10040421 ◽

2021 ◽

Vol 10 (4) ◽

pp. 421

Author(s):

Ebanja Joseph Ebwanga ◽

Stephen Mbigha Ghogomu ◽

Jan Paeshuyse

Keyword(s):

African Swine Fever Virus ◽

Central Africa ◽

African Swine Fever ◽

Fever Virus ◽

Economic Losses ◽

Future Research ◽

Genotype I ◽

Traditional System ◽

Pork Industry ◽

Domestic Cycle

African swine fever (ASF) is a hemorrhagic contagious porcine disease caused by the African swine fever virus. The disease poses enormous problems to the pork industry with pig mortality ranging from 30% to 100%, depending on the virulence of the virus circulating. Cameroon, situated in Central Africa is one of the countries in which the African swine fever virus (ASFV) has been endemic since its first outbreak in 1982. The disease is a major problem to the pig industry causing huge economic losses. A clear and concise review on ASF in Cameroon relating to the entry and current genotype of the virus, epidemiology, pathogenesis and economic impact is lacking. A thorough literature search revealed: (1) The virus entered the country in 1982 and caused the death of 80% of the pigs. (2) All isolates belong to serogroup I and only Genotype I is circulating in Cameroon principally in the domestic cycle as there are neither soft ticks nor warthog in the pig production regions sampled. (3) 70% of the pig farmers are involved in the traditional system of production with local and hybrid breeds of pigs with minimal input. (4) The country is endemic to the virus with huge economic losses. (5) So far, very little research has been effected on ASFV in Cameroon. This review gives a detailed overview of the situation of African swine fever virus (ASFV) in the country along with potential avenues for future research into ASFV in Cameroon.

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Short and Long-read Sequencing Survey of the Dynamic Transcriptomes of African Swine Fever Virus and its Host

10.1101/2020.02.27.967695 ◽

2020 ◽

Author(s):

Ferenc Olasz ◽

Dóra Tombácz ◽

Gábor Torma ◽

Zsolt Csabai ◽

Norbert Moldován ◽

...

Keyword(s):

African Swine Fever Virus ◽

African Swine Fever ◽

Fever Virus ◽

Economic Losses ◽

Swine Industry ◽

Rna Molecules ◽

Oxford Nanopore ◽

Combined Use ◽

Long Read ◽

Low Coverage

AbstractAfrican swine fever virus (ASFV) is an important animal pathogen causing substantial economic losses in the swine industry globally. At present, little is known about the molecular biology of ASFV, including its transcriptome organization. In this study, we applied cutting-edge sequencing approaches, namely the Illumina short-read sequencing (SRS) and the Oxford Nanopore Technologies long-read sequencing (LRS) techniques, together with several library preparation chemistries to analyze the ASFV dynamic transcriptome. SRS can generate a large amount of high-precision sequencing reads, but it is inefficient for identifying long RNA molecules, transcript isoforms and overlapping transcripts. LRS can overcome these limitations, but this approach also has shortcomings, such as its high error rate and the low coverage. Amplification-based LRS techniques produce relatively high read counts but also high levels of spurious transcripts, whereas the non-amplified cDNA and direct RNA sequencing techniques are more precise but achieve lower throughput. The drawbacks of the various technologies can be circumvented by the combined use of these approaches.

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