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Article ; Online: Isolation of Porcine Bone Marrow Cells and Generation of Recombinant African Swine Fever Viruses.

Rathakrishnan, Anusyah / Reis, Ana Luisa / Moffat, Katy / Dixon, Linda K

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2503, Page(s) 73–94

Abstract: Genetic manipulation of ASFV has been increasingly used not only for the development of live attenuated vaccines but also as an indispensable tool to further our understanding of the virus-host interactions. Here we present methods for isolation of ... ...

Abstract	Genetic manipulation of ASFV has been increasingly used not only for the development of live attenuated vaccines but also as an indispensable tool to further our understanding of the virus-host interactions. Here we present methods for isolation of porcine bone marrow cells and purification of recombinant ASFV using both chromogenic and fluorescent reporters. We also describe in detail a newly developed method to purify genetically modified ASFV using fluorescence-activated cell sorting (FACS).
MeSH term(s)	African Swine Fever/prevention & control ; African Swine Fever Virus/genetics ; Animals ; Bone Marrow Cells ; Swine ; Vaccines, Attenuated ; Viral Proteins/genetics
Chemical Substances	Vaccines, Attenuated ; Viral Proteins
Language	English
Publishing date	2022-05-16
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2333-6_5
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Deletion of the gene for the African swine fever virus BCL-2 family member A179L increases virus uptake and apoptosis but decreases virus spread in macrophages and reduces virulence in pigs.

Reis, Ana Luisa / Rathakrishnan, Anusyah / Goulding, Leah V / Barber, Claire / Goatley, Lynnette C / Dixon, Linda K

Journal of virology

2023 Volume 97, Issue 10, Page(s) e0110623

Abstract: Importance: African swine fever virus (ASFV) causes a lethal disease of pigs with high economic impact in affected countries in Africa, Europe, and Asia. The virus encodes proteins that inhibit host antiviral defenses, including the type I interferon ... ...

Abstract	Importance: African swine fever virus (ASFV) causes a lethal disease of pigs with high economic impact in affected countries in Africa, Europe, and Asia. The virus encodes proteins that inhibit host antiviral defenses, including the type I interferon response. Host cells also activate cell death through a process called apoptosis to limit virus replication. We showed that the ASFV A179L protein, a BCL-2 family apoptosis inhibitor, is important in reducing apoptosis in infected cells since deletion of this gene increased cell death and reduced virus replication in cells infected with the A179L gene-deleted virus. Pigs immunized with the BeninΔA179L virus showed no clinical signs and a weak immune response but were not protected from infection with the deadly parental virus. The results show an important role for the A179L protein in virus replication in macrophages and virulence in pigs and suggest manipulation of apoptosis as a possible route to control infection.
MeSH term(s)	Animals ; African Swine Fever/virology ; African Swine Fever Virus/genetics ; Apoptosis ; Gene Deletion ; Macrophages/virology ; Proto-Oncogene Proteins c-bcl-2/deficiency ; Proto-Oncogene Proteins c-bcl-2/genetics ; Swine/virology ; Virulence/genetics ; Virus Replication ; Apoptosis Regulatory Proteins/deficiency ; Apoptosis Regulatory Proteins/genetics ; Viral Proteins/genetics
Chemical Substances	Proto-Oncogene Proteins c-bcl-2 ; A179L protein, African Swine Fever Virus ; Apoptosis Regulatory Proteins ; Viral Proteins
Language	English
Publishing date	2023-10-05
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80174-4
ISSN	1098-5514 ; 0022-538X
ISSN (online)	1098-5514
ISSN	0022-538X
DOI	10.1128/jvi.01106-23
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Article ; Online: Wild boar visits to commercial pig farms in southwest England: implications for disease transmission

Bacigalupo, Sonny A. / Dixon, Linda K. / Gubbins, Simon / Kucharski, Adam J. / Drewe, Julian A.

Eur J Wildl Res. 2022 Dec., v. 68, no. 6 p.69-69

2022

Abstract: Contact between wild animals and farmed livestock may result in disease transmission with huge financial, welfare and ethical consequences. Conflicts between people and wildlife can also arise when species such as wild boar (Sus scrofa) consume crops or ... ...

Abstract	Contact between wild animals and farmed livestock may result in disease transmission with huge financial, welfare and ethical consequences. Conflicts between people and wildlife can also arise when species such as wild boar (Sus scrofa) consume crops or dig up pasture. This is a relatively recent problem in England where wild boar populations have become re-established in the last 20 years following a 500-year absence. The aim of this pilot study was to determine if and how often free-living wild boar visited two commercial pig farms near the Forest of Dean in southwest England. We placed 20 motion-sensitive camera traps at potential entry points to, and trails surrounding, the perimeter of two farmyards housing domestic pigs between August 2019 and February 2021, covering a total of 6030 trap nights. Forty wild boar detections were recorded on one farm spread across 27 nights, with a median (range) of 1 (0 to 7) night of wild boar activity per calendar month. Most of these wild boar detections occurred between ten and twenty metres of housed domestic pigs. No wild boar was detected at the other farm. These results confirm wild boar do visit commercial pig farms, and therefore, there is potential for contact and pathogen exchange between wild boar and domestic pigs. The visitation rates derived from this study could be used to parameterise disease transmission models of pathogens common to domestic pigs and wild boars, such as the African swine fever virus, and subsequently to develop mitigation strategies to reduce unwanted contacts.
Keywords	African swine fever virus ; Sus scrofa ; cameras ; disease transmission ; ethics ; farms ; forests ; pathogens ; people ; swine ; wild boars ; wildlife ; England
Language	English
Dates of publication	2022-12
Size	p. 69.
Publishing place	Springer Berlin Heidelberg
Document type	Article ; Online
ZDB-ID	2141660-6
ISSN	1439-0574 ; 1612-4642
ISSN (online)	1439-0574
ISSN	1612-4642
DOI	10.1007/s10344-022-01618-2
Database	NAL-Catalogue (AGRICOLA)

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Article: Wild boar visits to commercial pig farms in southwest England: implications for disease transmission.

Bacigalupo, Sonny A / Dixon, Linda K / Gubbins, Simon / Kucharski, Adam J / Drewe, Julian A

European journal of wildlife research

2022 Volume 68, Issue 6, Page(s) 69

Abstract	Contact between wild animals and farmed livestock may result in disease transmission with huge financial, welfare and ethical consequences. Conflicts between people and wildlife can also arise when species such as wild boar (Sus scrofa) consume crops or dig up pasture. This is a relatively recent problem in England where wild boar populations have become re-established in the last 20 years following a 500-year absence. The aim of this pilot study was to determine if and how often free-living wild boar visited two commercial pig farms near the Forest of Dean in southwest England. We placed 20 motion-sensitive camera traps at potential entry points to, and trails surrounding, the perimeter of two farmyards housing domestic pigs between August 2019 and February 2021, covering a total of 6030 trap nights. Forty wild boar detections were recorded on one farm spread across 27 nights, with a median (range) of 1 (0 to 7) night of wild boar activity per calendar month. Most of these wild boar detections occurred between ten and twenty metres of housed domestic pigs. No wild boar was detected at the other farm. These results confirm wild boar do visit commercial pig farms, and therefore, there is potential for contact and pathogen exchange between wild boar and domestic pigs. The visitation rates derived from this study could be used to parameterise disease transmission models of pathogens common to domestic pigs and wild boars, such as the African swine fever virus, and subsequently to develop mitigation strategies to reduce unwanted contacts.
Language	English
Publishing date	2022-10-04
Publishing country	Germany
Document type	Journal Article
ZDB-ID	2141660-6
ISSN	1439-0574 ; 1612-4642
ISSN (online)	1439-0574
ISSN	1612-4642
DOI	10.1007/s10344-022-01618-2
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Full genome sequence analysis of African swine fever virus isolates from Cameroon.

Goatley, Lynnette C / Freimanis, Graham / Tennakoon, Chandana / Foster, Thomas J / Quershi, Mehnaz / Dixon, Linda K / Batten, Carrie / Forth, Jan Hendrik / Wade, Abel / Netherton, Christopher

PloS one

2024 Volume 19, Issue 3, Page(s) e0293049

Abstract: African swine fever (ASF) is a devastating disease of domestic pigs that has spread across the globe since its introduction into Georgia in 2007. The etiological agent is a large double-stranded DNA virus with a genome of 170 to 180 kb in length ... ...

Abstract	African swine fever (ASF) is a devastating disease of domestic pigs that has spread across the globe since its introduction into Georgia in 2007. The etiological agent is a large double-stranded DNA virus with a genome of 170 to 180 kb in length depending on the isolate. Much of the differences in genome length between isolates are due to variations in the copy number of five different multigene families that are encoded in repetitive regions that are towards the termini of the covalently closed ends of the genome. Molecular epidemiology of African swine fever virus (ASFV) is primarily based on Sanger sequencing of a few conserved and variable regions, but due to the stability of the dsDNA genome changes in the variable regions occur relatively slowly. Observations in Europe and Asia have shown that changes in other genetic loci can occur and that this could be useful in molecular tracking. ASFV has been circulating in Western Africa for at least forty years. It is therefore reasonable to assume that changes may have accumulated in regions of the genome other than the standard targets over the years. At present only one full genome sequence is available for an isolate from Western Africa, that of a highly virulent isolate collected from Benin during an outbreak in 1997. In Cameroon, ASFV was first reported in 1981 and outbreaks have been reported to the present day and is considered endemic. Here we report three full genome sequences from Cameroon isolates of 1982, 1994 and 2018 outbreaks and identify novel single nucleotide polymorphisms and insertion-deletions that may prove useful for molecular epidemiology studies in Western Africa and beyond.
MeSH term(s)	Swine ; Animals ; African Swine Fever Virus ; African Swine Fever/epidemiology ; Cameroon/epidemiology ; Sus scrofa/genetics ; Sequence Analysis ; Sequence Analysis, DNA
Language	English
Publishing date	2024-03-21
Publishing country	United States
Document type	Journal Article
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0293049
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Article: Deletion of the K145R and DP148R Genes from the Virulent ASFV Georgia 2007/1 Isolate Delays the Onset, but Does Not Reduce Severity, of Clinical Signs in Infected Pigs

Rathakrishnan, Anusyah / Reis, Ana L. / Goatley, Lynnette C. / Moffat, Katy / Dixon, Linda K.

Viruses. 2021 July 28, v. 13, no. 8

2021

Abstract: African swine fever virus causes a frequently fatal disease of domestic pigs and wild boar that has a high economic impact across 3 continents. The large double-stranded DNA genome codes for approximately 160 proteins. Many of these have unknown ... ...

Abstract	African swine fever virus causes a frequently fatal disease of domestic pigs and wild boar that has a high economic impact across 3 continents. The large double-stranded DNA genome codes for approximately 160 proteins. Many of these have unknown functions and this hinders our understanding of the virus and host interactions. The purpose of the study was to evaluate the role of two virus proteins, K145R and DP148R, in virus replication in macrophages and virulence in pigs. To do this, the DP148R gene, alone or in combination with the K145R gene, was deleted from the virulent genotype II Georgia 2007/1 isolate. Neither of these deletions reduced the ability of the viruses to replicate in porcine macrophages compared to the parental wild-type virus. Pigs infected with GeorgiaΔDP148R developed clinical and post-mortem signs and high viremia, typical of acute African swine fever, and were culled on day 6 post-infection. The additional deletion of the K145R gene delayed the onset of clinical signs and viremia in pigs by 3 days, but pigs showed signs of acute African swine fever and were culled on days 10 or 13 post-infection. The results show that the deletion of DP148R did not attenuate the genotype II Georgia 2007/1 isolate, contrary to the results obtained with the genotype I Benin97/1 isolate. Additional deletion of the K145R gene delayed clinical signs, but infected pigs reached the humane endpoint. The deletion of additional genes would be required to attenuate the virus.
Keywords	African swine fever ; African swine fever virus ; DNA ; economic impact ; genes ; genotype ; humane endpoints ; macrophages ; swine ; viremia ; virulence ; virus replication ; viruses ; wild boars ; Georgia
Language	English
Dates of publication	2021-0728
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v13081473
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Production of Recombinant African Swine Fever Viruses: Speeding Up the Process.

Rathakrishnan, Anusyah / Moffat, Katy / Reis, Ana Luisa / Dixon, Linda K

Viruses

2020 Volume 12, Issue 6

Abstract: African swine fever (ASF) is a devastating disease in pigs, with no vaccines for control. The genetic manipulation of African swine fever virus (ASFV) is often tedious and time consuming. Here, we describe a method to manipulate the virus genome to ... ...

Abstract	African swine fever (ASF) is a devastating disease in pigs, with no vaccines for control. The genetic manipulation of African swine fever virus (ASFV) is often tedious and time consuming. Here, we describe a method to manipulate the virus genome to produce gene deletion viruses in a much-reduced time. This method combines the conventional homologous recombination with fluorescent-activated cells sorting (FACS), to isolate and purify viruses expressing fluorescent reporter genes. With three rounds of single cell isolation via FACS and two rounds of limiting dilution, we deleted two additional genes, EP153R and EP402R, from Benin 97/1 ASFV lacking the DP148R gene. By combining different fluorescent markers, this method has the potential to greatly facilitate studies on understanding ASFV gene functions and develop candidate live-attenuated vaccines.
MeSH term(s)	African Swine Fever/virology ; African Swine Fever Virus/genetics ; African Swine Fever Virus/immunology ; Animals ; Genetic Engineering ; Genome, Viral ; Recombination, Genetic ; Swine ; Viral Proteins/genetics ; Viral Proteins/immunology ; Viral Vaccines/genetics ; Viral Vaccines/immunology
Chemical Substances	Viral Proteins ; Viral Vaccines
Language	English
Publishing date	2020-06-05
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v12060615
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Deletion of the K145R and DP148R Genes from the Virulent ASFV Georgia 2007/1 Isolate Delays the Onset, but Does Not Reduce Severity, of Clinical Signs in Infected Pigs.

Rathakrishnan, Anusyah / Reis, Ana L / Goatley, Lynnette C / Moffat, Katy / Dixon, Linda K

Viruses

2021 Volume 13, Issue 8

Abstract	African swine fever virus causes a frequently fatal disease of domestic pigs and wild boar that has a high economic impact across 3 continents. The large double-stranded DNA genome codes for approximately 160 proteins. Many of these have unknown functions and this hinders our understanding of the virus and host interactions. The purpose of the study was to evaluate the role of two virus proteins, K145R and DP148R, in virus replication in macrophages and virulence in pigs. To do this, the DP148R gene, alone or in combination with the K145R gene, was deleted from the virulent genotype II Georgia 2007/1 isolate. Neither of these deletions reduced the ability of the viruses to replicate in porcine macrophages compared to the parental wild-type virus. Pigs infected with GeorgiaΔDP148R developed clinical and post-mortem signs and high viremia, typical of acute African swine fever, and were culled on day 6 post-infection. The additional deletion of the K145R gene delayed the onset of clinical signs and viremia in pigs by 3 days, but pigs showed signs of acute African swine fever and were culled on days 10 or 13 post-infection. The results show that the deletion of DP148R did not attenuate the genotype II Georgia 2007/1 isolate, contrary to the results obtained with the genotype I Benin97/1 isolate. Additional deletion of the K145R gene delayed clinical signs, but infected pigs reached the humane endpoint. The deletion of additional genes would be required to attenuate the virus.
MeSH term(s)	African Swine Fever/virology ; African Swine Fever Virus/genetics ; African Swine Fever Virus/pathogenicity ; African Swine Fever Virus/physiology ; Animals ; Gene Deletion ; Macrophages/virology ; Swine ; Viral Proteins/genetics ; Viral Proteins/metabolism ; Virulence ; Virus Replication
Chemical Substances	Viral Proteins
Language	English
Publishing date	2021-07-28
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v13081473
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: The importance of fine-scale predictors of wild boar habitat use in an isolated population.

Bacigalupo, Sonny A / Chang, Yu-Mei / Dixon, Linda K / Gubbins, Simon / Kucharski, Adam J / Drewe, Julian A

Ecology and evolution

2022 Volume 12, Issue 6, Page(s) e9031

Abstract: Predicting the likelihood of wildlife presence at potential wildlife-livestock interfaces is challenging. These interfaces are usually relatively small geographical areas where landscapes show large variation over small distances. Models of wildlife ... ...

Abstract	Predicting the likelihood of wildlife presence at potential wildlife-livestock interfaces is challenging. These interfaces are usually relatively small geographical areas where landscapes show large variation over small distances. Models of wildlife distribution based on coarse data over wide geographical ranges may not be representative of these interfaces. High-resolution data can help identify fine-scale predictors of wildlife habitat use at a local scale and provide more accurate predictions of species habitat use. These data may be used to inform knowledge of interface risks, such as disease transmission between wildlife and livestock, or human-wildlife conflict.This study uses fine-scale habitat use data from wild boar (
Language	English
Publishing date	2022-06-22
Publishing country	England
Document type	Journal Article
ZDB-ID	2635675-2
ISSN	2045-7758
ISSN	2045-7758
DOI	10.1002/ece3.9031
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Article: The Genetics of Life and Death: Virus-Host Interactions Underpinning Resistance to African Swine Fever, a Viral Hemorrhagic Disease.

Netherton, Christopher L / Connell, Samuel / Benfield, Camilla T O / Dixon, Linda K

Frontiers in genetics

2019 Volume 10, Page(s) 402

Abstract: Pathogen transmission from wildlife hosts to genetically distinct species is a major driver of disease emergence. African swine fever virus (ASFV) persists in sub-Saharan Africa through a sylvatic cycle between warthogs and soft ticks that infest their ... ...

Abstract	Pathogen transmission from wildlife hosts to genetically distinct species is a major driver of disease emergence. African swine fever virus (ASFV) persists in sub-Saharan Africa through a sylvatic cycle between warthogs and soft ticks that infest their burrows. The virus does not cause disease in these animals, however transmission of the virus to domestic pigs or wild boar causes a hemorrhagic fever that is invariably fatal. ASFV transmits readily between domestic pigs and causes economic hardship in areas where it is endemic. The virus is also a significant transboundary pathogen that has become established in Eastern Europe, and has recently appeared in China increasing the risk of an introduction of the disease to other pig producing centers. Although a DNA genome mitigates against rapid adaptation of the virus to new hosts, extended epidemics of African swine fever (ASF) can lead to the emergence of viruses with reduced virulence. Attenuation in the field leads to large deletions of genetic material encoding genes involved in modulating host immune responses. Therefore resistance to disease and tolerance of ASFV replication can be dependent on both virus and host factors. Here we describe the different virus-host interfaces and discuss progress toward understanding the genetic determinants of disease outcome after infection with ASFV.
Language	English
Publishing date	2019-05-03
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2606823-0
ISSN	1664-8021
ISSN	1664-8021
DOI	10.3389/fgene.2019.00402
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