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Prof. Graham Belsham
University of Copenhagen, Department of Veterinary and Animal Sciences

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0 Molecular Virology
0 RNA structures
0 Picornaviruses
0 RNA viruses
0 Virus biology

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Journal article
Published: 16 July 2021 in Biosafety and Health
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Foot-and-mouth disease virus (FMDV), swine vesicular disease virus (SVDV) and classical swine fever virus (CSFV) all cause important animal diseases. FMDV affects many different cloven-hoofed animals, whereas SVDV and CSFV are restricted to domestic and feral pigs together with wild boar. Europe is normally free of these diseases, but occasionally outbreaks happen, which can cause huge economic losses. Handling of these viruses, in particular FMDV and CSFV, is only allowed within high containment laboratories and stables. Periodically such facilities need to be decommissioned for repair or closing down, which is done by cleaning and chemical disinfection, followed by fumigation due to residual risk from virus on surfaces in inaccessible places. However, building materials in older laboratories or stables that have housed infected animals may not be well-suited for fumigation. Heat treatment is another way of inactivating viruses. In this study, we have determined the survival of infectivity in air-dried virus samples on glass and plastic surfaces incubated at room temperature or heated to 70 °C for 1, 2, 3, 5 and 7 days. Each of the tested viruses was inactivated to below the limit of detection after 24 or 48 h of incubation at 70 °C; in contrast, some of these viruses were still infectious after 7 days of incubation at room temperature. This study provides important information that can be used in relation to decontamination of buildings and in risk-assessments.

ACS Style

Thea Kristensen; Graham J. Belsham; Kirsten Tjørnehøj. Heat inactivation of foot-and-mouth disease virus, swine vesicular disease virus and classical swine fever virus when air-dried on plastic and glass surfaces. Biosafety and Health 2021, 1 .

AMA Style

Thea Kristensen, Graham J. Belsham, Kirsten Tjørnehøj. Heat inactivation of foot-and-mouth disease virus, swine vesicular disease virus and classical swine fever virus when air-dried on plastic and glass surfaces. Biosafety and Health. 2021; ():1.

Chicago/Turabian Style

Thea Kristensen; Graham J. Belsham; Kirsten Tjørnehøj. 2021. "Heat inactivation of foot-and-mouth disease virus, swine vesicular disease virus and classical swine fever virus when air-dried on plastic and glass surfaces." Biosafety and Health , no. : 1.

Journal article
Published: 04 June 2021 in Viruses
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Bat species worldwide are receiving increased attention for the discovery of emerging viruses, cross-species transmission, and zoonoses, as well as for characterizing virus infections specific to bats. In a previous study, we investigated the presence of coronaviruses in faecal samples from bats at different locations in Denmark, and made phylogenies based on short, partial ORF1b sequences. In this study, selected samples containing bat coronaviruses from three different bat species were analysed, using a non-targeted approach of next-generation sequencing. From the resulting metagenomics data, we assembled full-genome sequences of seven distinct alphacoronaviruses, three astroviruses, and a polyomavirus, as well as partial genome sequences of rotavirus H and caliciviruses, from the different bat species. Comparisons to published sequences indicate that the bat alphacoronaviruses belong to three different subgenera—i.e., Pedacovirus, Nyctacovirus, and Myotacovirus—that the astroviruses may be new species in the genus Mamastrovirus, and that the polyomavirus could also be a new species, but unassigned to a genus. Furthermore, several viruses of invertebrates—including two Rhopalosiphum padi (aphid) viruses and a Kadipiro virus—present in the faecal material were assembled. Interestingly, this is the first detection in Europe of a Kadipiro virus.

ACS Style

Christina Lazov; Graham Belsham; Anette Bøtner; Thomas Rasmussen. Full-Genome Sequences of Alphacoronaviruses and Astroviruses from Myotis and Pipistrelle Bats in Denmark. Viruses 2021, 13, 1073 .

AMA Style

Christina Lazov, Graham Belsham, Anette Bøtner, Thomas Rasmussen. Full-Genome Sequences of Alphacoronaviruses and Astroviruses from Myotis and Pipistrelle Bats in Denmark. Viruses. 2021; 13 (6):1073.

Chicago/Turabian Style

Christina Lazov; Graham Belsham; Anette Bøtner; Thomas Rasmussen. 2021. "Full-Genome Sequences of Alphacoronaviruses and Astroviruses from Myotis and Pipistrelle Bats in Denmark." Viruses 13, no. 6: 1073.

Preprint content
Published: 07 May 2021
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Mink, on a farm with about 15,000 animals, became infected with SARS-CoV-2. Over 75% of tested animals were positive for SARS-CoV-2 RNA in throat swabs and 100% of tested animals were seropositive. The virus responsible had a deletion of nucleotides encoding residues H69 and V70 within the spike protein gene. The infected mink recovered and after free-testing of the mink, the animals remained seropositive. During follow-up studies, after a period of more than 2 months without virus detection, over 75% of tested animals scored positive again for SARS-CoV-2 RNA. Whole genome sequencing showed that the virus circulating during this re-infection was most closely related to the virus identified in the first outbreak on this farm but additional sequence changes had occurred. Animals had much higher levels of anti-SARS-CoV-2 antibodies after re-infection than at free-testing. Thus, following recovery from an initial infection, seropositive mink rapidly became susceptible to re-infection by SARS-CoV-2.

ACS Style

Thomas Bruun Rasmussen; Jannik Fonager; Charlotte Sværke Jørgensen; Ria Lassaunière; Anne Sofie Hammer; Michelle Lauge Quaade; Anette Boklund; Louise Lohse; Bertel Strandbygaard; Morten Rasmussen; Thomas Yssing Michaelsen; Sten Mortensen; Anders Fomsgaard; Graham J. Belsham; Anette Bøtner. Infection, recovery and re-infection of farmed mink with SARS-CoV-2. 2021, 1 .

AMA Style

Thomas Bruun Rasmussen, Jannik Fonager, Charlotte Sværke Jørgensen, Ria Lassaunière, Anne Sofie Hammer, Michelle Lauge Quaade, Anette Boklund, Louise Lohse, Bertel Strandbygaard, Morten Rasmussen, Thomas Yssing Michaelsen, Sten Mortensen, Anders Fomsgaard, Graham J. Belsham, Anette Bøtner. Infection, recovery and re-infection of farmed mink with SARS-CoV-2. . 2021; ():1.

Chicago/Turabian Style

Thomas Bruun Rasmussen; Jannik Fonager; Charlotte Sværke Jørgensen; Ria Lassaunière; Anne Sofie Hammer; Michelle Lauge Quaade; Anette Boklund; Louise Lohse; Bertel Strandbygaard; Morten Rasmussen; Thomas Yssing Michaelsen; Sten Mortensen; Anders Fomsgaard; Graham J. Belsham; Anette Bøtner. 2021. "Infection, recovery and re-infection of farmed mink with SARS-CoV-2." , no. : 1.

Rapid communication
Published: 29 April 2021 in Transboundary and Emerging Diseases
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The presence of foot‐and‐mouth disease virus (FMDV) of the O/ME‐SA/Ind‐2001e sublineage within Pakistan was initially detected in two samples collected during 2019. Analysis of further serotype O FMDVs responsible for disease outbreaks in 2019‐2020 in the country has now identified the spread of this sublineage to 10 districts within two separate provinces in North‐Eastern and North‐Western Pakistan. Phylogenetic analysis indicates that these viruses are closely related to those circulating in Bhutan, Nepal and India. The VP1 coding sequences of these viruses from Pakistan belong to three distinct clusters, which may indicate multiple introductions of this virus sublineage, although the routes of introduction are unknown. Vaccine matching studies against O1 Manisa, O 3039 and O TUR/5/2009 support the suitability of existing vaccine strains to control current field outbreaks, but further studies are warranted to monitor the spread and evolution of the O/ME‐SA/Ind‐2001e sublineage in the region.

ACS Style

Syed M. Jamal; Salman Khan; Nick J. Knowles; Jemma Wadsworth; Hayley M. Hicks; Valérie Mioulet; Abdelghani Bin‐Tarif; Anna B. Ludi; Syed Asad Ali Shah; Muhammad Abubakar; Shumaila Manzoor; Muhammad Afzal; Michael Eschbaumer; Donald P. King; Graham J. Belsham. Foot‐and‐mouth disease viruses of the O/ME‐SA/Ind‐2001e sublineage in Pakistan. Transboundary and Emerging Diseases 2021, 1 .

AMA Style

Syed M. Jamal, Salman Khan, Nick J. Knowles, Jemma Wadsworth, Hayley M. Hicks, Valérie Mioulet, Abdelghani Bin‐Tarif, Anna B. Ludi, Syed Asad Ali Shah, Muhammad Abubakar, Shumaila Manzoor, Muhammad Afzal, Michael Eschbaumer, Donald P. King, Graham J. Belsham. Foot‐and‐mouth disease viruses of the O/ME‐SA/Ind‐2001e sublineage in Pakistan. Transboundary and Emerging Diseases. 2021; ():1.

Chicago/Turabian Style

Syed M. Jamal; Salman Khan; Nick J. Knowles; Jemma Wadsworth; Hayley M. Hicks; Valérie Mioulet; Abdelghani Bin‐Tarif; Anna B. Ludi; Syed Asad Ali Shah; Muhammad Abubakar; Shumaila Manzoor; Muhammad Afzal; Michael Eschbaumer; Donald P. King; Graham J. Belsham. 2021. "Foot‐and‐mouth disease viruses of the O/ME‐SA/Ind‐2001e sublineage in Pakistan." Transboundary and Emerging Diseases , no. : 1.

Review
Published: 11 March 2021 in Viruses
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Picornaviruses are comprised of a positive-sense RNA genome surrounded by a protein shell (or capsid). They are ubiquitous in vertebrates and cause a wide range of important human and animal diseases. The genome encodes a single large polyprotein that is processed to structural (capsid) and non-structural proteins. The non-structural proteins have key functions within the viral replication complex. Some, such as 3Dpol (the RNA dependent RNA polymerase) have conserved functions and participate directly in replicating the viral genome, whereas others, such as 3A, have accessory roles. The 3A proteins are highly divergent across the Picornaviridae and have specific roles both within and outside of the replication complex, which differ between the different genera. These roles include subverting host proteins to generate replication organelles and inhibition of cellular functions (such as protein secretion) to influence virus replication efficiency and the host response to infection. In addition, 3A proteins are associated with the determination of host range. However, recent observations have challenged some of the roles assigned to 3A and suggest that other viral proteins may carry them out. In this review, we revisit the roles of 3A in the picornavirus life cycle. The 3AB precursor and mature 3A have distinct functions during viral replication and, therefore, we have also included discussion of some of the roles assigned to 3AB.

ACS Style

Terry Jackson; Graham Belsham. Picornaviruses: A View from 3A. Viruses 2021, 13, 456 .

AMA Style

Terry Jackson, Graham Belsham. Picornaviruses: A View from 3A. Viruses. 2021; 13 (3):456.

Chicago/Turabian Style

Terry Jackson; Graham Belsham. 2021. "Picornaviruses: A View from 3A." Viruses 13, no. 3: 456.

Journal article
Published: 12 January 2021 in Animals
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SARS-CoV-2 infection is the cause of COVID-19 in humans. In April 2020, SARS-CoV-2 infection in farmed mink (Neovision vision) occurred in the Netherlands. The first outbreaks in Denmark were detected in June 2020 in three farms. A steep increase in the number of infected farms occurred from September and onwards. Here, we describe prevalence data collected from 215 infected mink farms to characterize spread and impact of disease in infected farms. In one third of the farms, no clinical signs were observed. In farms with clinical signs, decreased feed intake, increased mortality and respiratory symptoms were most frequently observed, during a limited time period (median of 11 days). In 65% and 69% of farms, virus and sero-conversion, respectively, were detected in 100% of sampled animals at the first sampling. SARS-CoV-2 was detected, at low levels, in air samples collected close to the mink, on mink fur, on flies, on the foot of a seagull, and in gutter water, but not in feed. Some dogs and cats from infected farms tested positive for the virus. Chickens, rabbits, and horses sampled on a few farms, and wildlife sampled in the vicinity of the infected farms did not test positive for SARS-CoV-2. Thus, mink are highly susceptible to infection by SARS-CoV-2, but routes of transmission between farms, other than by direct human contact, are unclear.

ACS Style

Anette Boklund; Anne Sofie Hammer; Michelle Lauge Quaade; Thomas Bruun Rasmussen; Louise Lohse; Bertel Strandbygaard; Charlotte Sværke Jørgensen; Ann Sofie Olesen; Freja Broe Hjerpe; Heidi Huus Petersen; Tim Kåre Jensen; Sten Mortensen; Francisco F. Calvo-Artavia; Stine Kjær Lefèvre; Søren Saxmose Nielsen; Tariq Halasa; Graham J. Belsham; Anette Bøtner. SARS-CoV-2 in Danish Mink Farms: Course of the Epidemic and a Descriptive Analysis of the Outbreaks in 2020. Animals 2021, 11, 164 .

AMA Style

Anette Boklund, Anne Sofie Hammer, Michelle Lauge Quaade, Thomas Bruun Rasmussen, Louise Lohse, Bertel Strandbygaard, Charlotte Sværke Jørgensen, Ann Sofie Olesen, Freja Broe Hjerpe, Heidi Huus Petersen, Tim Kåre Jensen, Sten Mortensen, Francisco F. Calvo-Artavia, Stine Kjær Lefèvre, Søren Saxmose Nielsen, Tariq Halasa, Graham J. Belsham, Anette Bøtner. SARS-CoV-2 in Danish Mink Farms: Course of the Epidemic and a Descriptive Analysis of the Outbreaks in 2020. Animals. 2021; 11 (1):164.

Chicago/Turabian Style

Anette Boklund; Anne Sofie Hammer; Michelle Lauge Quaade; Thomas Bruun Rasmussen; Louise Lohse; Bertel Strandbygaard; Charlotte Sværke Jørgensen; Ann Sofie Olesen; Freja Broe Hjerpe; Heidi Huus Petersen; Tim Kåre Jensen; Sten Mortensen; Francisco F. Calvo-Artavia; Stine Kjær Lefèvre; Søren Saxmose Nielsen; Tariq Halasa; Graham J. Belsham; Anette Bøtner. 2021. "SARS-CoV-2 in Danish Mink Farms: Course of the Epidemic and a Descriptive Analysis of the Outbreaks in 2020." Animals 11, no. 1: 164.

Journal article
Published: 01 November 2020 in Journal of General Virology
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Border disease virus (BDV) envelope glycoprotein E2 is required for entry into cells and is a determinant of host tropism for sheep and pig cells. Here, we describe adaptive changes in the BDV E2 protein that modify virus replication in pig cells. To achieve this, two BDV isolates, initially collected from a pig and a sheep on the same farm, were passaged in primary sheep and pig cells in parallel with a rescued variant of the pig virus derived from a cloned full-length BDV cDNA. The pig isolate and the rescued virus shared the same amino acid sequence, but the sheep isolate differed at ten residues, including two substitutions in E2 (K771E and Y925H). During serial passage in cells, the viruses displayed clear selectivity for growth in sheep cells; only the cDNA-derived virus adapted to grow in pig cells. Sequencing revealed an amino acid substitution (Q739R) in the E2 domain DA of this rescued virus. Adaptation at the same residue (Q739K/Q739R) was also observed after passaging of the pig isolate in sheep cells. Use of reverse genetics confirmed that changing residue Q739 to R or K (each positively charged) was sufficient to achieve adaptation to pig cells. Furthermore, this change in host tropism was suppressed if Q739R was combined with K771E. Another substitution (Q728R), conferring an additional positive charge, acquired during passaging, restored the growth of the Q739R/K771E variant. Overall, this study provided evidence that specific, positively charged, residues in the E2 domain DA are crucial for pig-cell tropism of BDV.

ACS Style

Ulrik Fahnøe; Yu Deng; Nana A. Davids; Louise Lohse; Jens Bukh; Graham J. Belsham; Thomas Bruun Rasmussen. Identification of specific amino acid residues in the border disease virus glycoprotein E2 that modify virus growth in pig cells but not in sheep cells. Journal of General Virology 2020, 101, 1170 -1181.

AMA Style

Ulrik Fahnøe, Yu Deng, Nana A. Davids, Louise Lohse, Jens Bukh, Graham J. Belsham, Thomas Bruun Rasmussen. Identification of specific amino acid residues in the border disease virus glycoprotein E2 that modify virus growth in pig cells but not in sheep cells. Journal of General Virology. 2020; 101 (11):1170-1181.

Chicago/Turabian Style

Ulrik Fahnøe; Yu Deng; Nana A. Davids; Louise Lohse; Jens Bukh; Graham J. Belsham; Thomas Bruun Rasmussen. 2020. "Identification of specific amino acid residues in the border disease virus glycoprotein E2 that modify virus growth in pig cells but not in sheep cells." Journal of General Virology 101, no. 11: 1170-1181.

Journal article
Published: 15 September 2020 in Journal of Virology
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The surface-exposed E2 protein of classical swine fever virus is required for its interaction with host cells. A short motif within this protein varies between strains of different virulence. The importance of two particular amino acid residues in determining the properties of a highly virulent strain of the virus has been analyzed. Each of the different viruses tested proved highly virulent, but one of them produced earlier, but not more severe, disease. By analyzing the virus genomes present within infected pigs, it was found that the viruses which replicated within inoculated animals were only a subset of those within the virus inoculum. Furthermore, following contact transmission, it was shown that a very restricted set of viruses had transferred between animals. There were no significant differences in the virus populations present in various tissues of the infected animals. These results indicate mechanisms of virus population change during transmission between animals.

ACS Style

Camille Melissa Johnston; Ulrik Fahnøe; Louise Lohse; Jens Bukh; Graham J. Belsham; Thomas Bruun Rasmussen. Analysis of Virus Population Profiles within Pigs Infected with Virulent Classical Swine Fever Viruses: Evidence for Bottlenecks in Transmission but Absence of Tissue-Specific Virus Variants. Journal of Virology 2020, 94, 1 .

AMA Style

Camille Melissa Johnston, Ulrik Fahnøe, Louise Lohse, Jens Bukh, Graham J. Belsham, Thomas Bruun Rasmussen. Analysis of Virus Population Profiles within Pigs Infected with Virulent Classical Swine Fever Viruses: Evidence for Bottlenecks in Transmission but Absence of Tissue-Specific Virus Variants. Journal of Virology. 2020; 94 (19):1.

Chicago/Turabian Style

Camille Melissa Johnston; Ulrik Fahnøe; Louise Lohse; Jens Bukh; Graham J. Belsham; Thomas Bruun Rasmussen. 2020. "Analysis of Virus Population Profiles within Pigs Infected with Virulent Classical Swine Fever Viruses: Evidence for Bottlenecks in Transmission but Absence of Tissue-Specific Virus Variants." Journal of Virology 94, no. 19: 1.

Review
Published: 20 May 2020 in Acta Veterinaria Scandinavica
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Foot-and-mouth disease (FMD) remains one of the most economically important infectious diseases of production animals. Six (out of 7 that have been identified) different serotypes of the FMD virus continue to circulate in different parts of the world. Within each serotype there is also extensive diversity as the virus constantly changes. Vaccines need to be “matched” to the outbreak strain, not just to the serotype, to confer protection. Vaccination has been used successfully to assist in the eradication of the disease from Europe but is no longer employed there unless outbreaks occur. Thus the animal population in Europe, as in North America, is fully susceptible to the virus if it is accidentally (or deliberately) introduced. Almost 3 billion doses of the vaccine are made each year to control the disease elsewhere. Current vaccines are produced from chemically inactivated virus that has to be grown, on a large scale, under high containment conditions. The vaccine efficiently prevents disease but the duration of immunity is rather limited (about 6 months) and vaccination does not provide sterile immunity or block the development of carriers. Furthermore, the vaccine is quite unstable and a cold chain needs to be maintained to preserve the efficacy of the vaccine. This can be a challenge in the parts of the world where the disease is endemic. There is a significant interest in developing improved vaccines and significant progress in this direction has been made using a variety of approaches. However, no alternative vaccines are yet available commercially. Improved disease control globally is clearly beneficial to all countries as it reduces the risk of virus incursions into disease free areas.

ACS Style

Graham J. Belsham. Towards improvements in foot-and-mouth disease vaccine performance. Acta Veterinaria Scandinavica 2020, 62, 1 -12.

AMA Style

Graham J. Belsham. Towards improvements in foot-and-mouth disease vaccine performance. Acta Veterinaria Scandinavica. 2020; 62 (1):1-12.

Chicago/Turabian Style

Graham J. Belsham. 2020. "Towards improvements in foot-and-mouth disease vaccine performance." Acta Veterinaria Scandinavica 62, no. 1: 1-12.

Review
Published: 09 March 2020 in Transboundary and Emerging Diseases
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Following its introduction into Georgia in 2007, African swine fever virus (ASFV) has become widespread on the European continent and in Asia. In many cases, the exact route of introduction into domestic pig herds cannot be determined, but most introductions are attributed to indirect virus transmission. In this review, we describe knowledge gained about different matrices that may allow introduction of the virus into pig herds. These matrices include uncooked pig meat, processed pig‐derived products, feed, matrices contaminated with the virus and blood‐feeding invertebrates. Knowledge gaps still exist and both field studies and laboratory research are needed to enhance understanding of the risks for ASFV introductions, especially via virus‐contaminated materials, including bedding and feed, and via blood‐feeding, flying insects. Knowledge obtained from such studies can be applied to epidemiological risk assessments for the different transmission routes. Such assessments can be utilized to help predict the most effective biosecurity and control strategies.

ACS Style

Ann Sofie Olesen; Graham J. Belsham; Thomas Bruun Rasmussen; Louise Lohse; René Bødker; Tariq Halasa; Anette Boklund; Anette Bøtner. Potential routes for indirect transmission of African swine fever virus into domestic pig herds. Transboundary and Emerging Diseases 2020, 67, 1472 -1484.

AMA Style

Ann Sofie Olesen, Graham J. Belsham, Thomas Bruun Rasmussen, Louise Lohse, René Bødker, Tariq Halasa, Anette Boklund, Anette Bøtner. Potential routes for indirect transmission of African swine fever virus into domestic pig herds. Transboundary and Emerging Diseases. 2020; 67 (4):1472-1484.

Chicago/Turabian Style

Ann Sofie Olesen; Graham J. Belsham; Thomas Bruun Rasmussen; Louise Lohse; René Bødker; Tariq Halasa; Anette Boklund; Anette Bøtner. 2020. "Potential routes for indirect transmission of African swine fever virus into domestic pig herds." Transboundary and Emerging Diseases 67, no. 4: 1472-1484.

Review article
Published: 29 February 2020 in Virus Research
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Understanding of the biology of foot-and-mouth disease virus (FMDV) has grown considerably since the nucleotide sequence of the viral RNA was determined. The ability to manipulate the intact genome and also to express specific parts of the genome individually has enabled detailed analyses of viral components, both RNA and protein. Such studies have identified the requirements for specific functional elements for virus replication and pathogenicity. Furthermore, information about the functions of individual virus proteins has enabled the rational design of cDNA cassettes to express non-infectious empty capsid particles that can induce protective immunity in the natural host animals and thus represent new vaccine candidates. Similarly, attempts to block specific virus activities using antiviral agents have also been performed. However, currently, only the well-established, chemically inactivated FMDV vaccines are commercially available and suitable for use to combat this important disease of livestock animals. These vaccines, despite certain shortcomings, have been used very successfully (e.g. in Europe) to control the disease but it still remains endemic in much of Africa, southern Asia and the Middle East. Hence there remains a significant risk of reintroduction of the disease into highly susceptible animal populations with enormous economic consequences.

ACS Style

Graham J. Belsham; Thea Kristensen; Terry Jackson. Foot-and-mouth disease virus: Prospects for using knowledge of virus biology to improve control of this continuing global threat. Virus Research 2020, 281, 197909 .

AMA Style

Graham J. Belsham, Thea Kristensen, Terry Jackson. Foot-and-mouth disease virus: Prospects for using knowledge of virus biology to improve control of this continuing global threat. Virus Research. 2020; 281 ():197909.

Chicago/Turabian Style

Graham J. Belsham; Thea Kristensen; Terry Jackson. 2020. "Foot-and-mouth disease virus: Prospects for using knowledge of virus biology to improve control of this continuing global threat." Virus Research 281, no. : 197909.

Original article
Published: 23 November 2019 in Transboundary and Emerging Diseases
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Phylogenetic studies on foot‐and‐mouth disease viruses (FMDVs) circulating in the West Eurasian region have largely focused on the genomic sequences encoding the structural proteins that determine the serotype. The present study has compared near complete genome sequences of FMDVs representative of the viruses that circulate in this region. The near complete genome sequences (ca. 7600nt) were generated from multiple overlapping RT‐PCR products. These amplicons were from FMDVs belonging to serotypes O, A and Asia‐1, including members of the O‐PanAsia‐II and the A‐Iran05 lineages, and of Group‐II and Group‐VII (Sindh‐08) within serotype Asia‐1, which are currently predominant and widespread in West Eurasia. These new sequences were analysed together with other sequences obtained from GenBank. Comparison of different regions of the FMDVs genomes revealed evidence for multiple, inter‐serotypic, recombination events between FMDVs belonging to the serotypes O, A and Asia‐1. It is concluded from the present study that dramatic changes in virus sequences can occur in the field through recombination between different FMDV genomes. These analyses provide information about the ancestry of the serotype O, A and Asia‐1 FMDVs that are currently circulating within the West Eurasian region.

ACS Style

Syed M. Jamal; Mohamad Hossein Nazem Shirazi; Fuat Özyörük; Ünal Parlak; Preben Normann; Graham J. Belsham. Evidence for multiple recombination events within foot‐and‐mouth disease viruses circulating in West Eurasia. Transboundary and Emerging Diseases 2019, 67, 979 -993.

AMA Style

Syed M. Jamal, Mohamad Hossein Nazem Shirazi, Fuat Özyörük, Ünal Parlak, Preben Normann, Graham J. Belsham. Evidence for multiple recombination events within foot‐and‐mouth disease viruses circulating in West Eurasia. Transboundary and Emerging Diseases. 2019; 67 (2):979-993.

Chicago/Turabian Style

Syed M. Jamal; Mohamad Hossein Nazem Shirazi; Fuat Özyörük; Ünal Parlak; Preben Normann; Graham J. Belsham. 2019. "Evidence for multiple recombination events within foot‐and‐mouth disease viruses circulating in West Eurasia." Transboundary and Emerging Diseases 67, no. 2: 979-993.

Journal article
Published: 10 October 2019 in Viruses
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Vaccines against classical swine fever have proven very effective in protecting pigs from this deadly disease. However, little is known about how vaccination impacts the selective pressures acting on the classical swine fever virus (CSFV). Here we use high-throughput sequencing of viral genomes to investigate evolutionary changes in virus populations following the challenge of naïve and vaccinated pigs with the highly virulent CSFV strain “Koslov”. The challenge inoculum contained an ensemble of closely related viral sequences, with three major haplotypes being present, termed A, B, and C. After the challenge, the viral haplotype A was preferentially located within the tonsils of naïve animals but was highly prevalent in the sera of all vaccinated animals. We find that the viral population structure in naïve pigs after infection is very similar to that in the original inoculum. In contrast, the viral population in vaccinated pigs, which only underwent transient low-level viremia, displayed several distinct changes including the emergence of 16 unique non-synonymous single nucleotide polymorphisms (SNPs) that were not detectable in the challenge inoculum. Further analysis showed a significant loss of heterogeneity and an increasing positive selection acting on the virus populations in the vaccinated pigs. We conclude that vaccination imposes a strong selective pressure on viruses that subsequently replicate within the vaccinated animal.

ACS Style

Ulrik Fahnøe; Anders Gorm Pedersen; Camille Melissa Johnston; Richard J. Orton; Dirk Höper; Martin Beer; Jens Bukh; Graham J. Belsham; Thomas Bruun Rasmussen. Virus Adaptation and Selection Following Challenge of Animals Vaccinated against Classical Swine Fever Virus. Viruses 2019, 11, 932 .

AMA Style

Ulrik Fahnøe, Anders Gorm Pedersen, Camille Melissa Johnston, Richard J. Orton, Dirk Höper, Martin Beer, Jens Bukh, Graham J. Belsham, Thomas Bruun Rasmussen. Virus Adaptation and Selection Following Challenge of Animals Vaccinated against Classical Swine Fever Virus. Viruses. 2019; 11 (10):932.

Chicago/Turabian Style

Ulrik Fahnøe; Anders Gorm Pedersen; Camille Melissa Johnston; Richard J. Orton; Dirk Höper; Martin Beer; Jens Bukh; Graham J. Belsham; Thomas Bruun Rasmussen. 2019. "Virus Adaptation and Selection Following Challenge of Animals Vaccinated against Classical Swine Fever Virus." Viruses 11, no. 10: 932.

Journal article
Published: 13 August 2019 in Scientific Reports
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The picornavirus family includes poliovirus (PV) (genus: enterovirus), human rhinoviruses (enterovirus) and foot-and-mouth disease virus (FMDV) (aphthovirus). These are responsible for important human and animal health concerns worldwide including poliomyelitis, the common cold and foot-and-mouth disease (FMD) respectively. In picornavirus particles, the positive-sense RNA genome (ca. 7–9 kb) is packaged within a protein shell (capsid) usually consisting of three surface exposed proteins, VP1, VP2 and VP3 plus the internal VP4, which are generated following cleavage of the capsid precursor by a virus-encoded protease. We have previously identified a motif near the C-terminus of FMDV VP1 that is required for capsid precursor processing. This motif is highly conserved among other picornaviruses, and is also likely to be important for their capsid precursor processing. We have now determined the plasticity of residues within this motif for virus infectivity and found an important interaction between FMDV residue VP1 R188 within this conserved motif and residue W129 in VP2 that is adjacent in the virus capsid. The FMDV (VP1 R188A) mutant virus has only been rescued with the secondary substitution VP2 W129R. This additional change compensates for the defect resulting from the VP1 R188A substitution and restored both capsid precursor processing and virus viability.

ACS Style

Thea Kristensen; Graham J. Belsham. Identification of plasticity and interactions of a highly conserved motif within a picornavirus capsid precursor required for virus infectivity. Scientific Reports 2019, 9, 1 -10.

AMA Style

Thea Kristensen, Graham J. Belsham. Identification of plasticity and interactions of a highly conserved motif within a picornavirus capsid precursor required for virus infectivity. Scientific Reports. 2019; 9 (1):1-10.

Chicago/Turabian Style

Thea Kristensen; Graham J. Belsham. 2019. "Identification of plasticity and interactions of a highly conserved motif within a picornavirus capsid precursor required for virus infectivity." Scientific Reports 9, no. 1: 1-10.

Journal article
Published: 15 May 2019 in Journal of Virology
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This study provides novel insights into the little-known structural determinants of the balance between thermal stability and instability in the capsid of foot-and-mouth disease virus and into the relationship between capsid stability and virus infectivity. The results provide new guidelines for the development of thermostabilized empty capsid-based recombinant vaccines against foot-and-mouth disease, one of the economically most important animal diseases worldwide.

ACS Style

Silvia López-Argüello; Verónica Rincón; Alicia Rodríguez-Huete; Encarnación Martínez-Salas; Graham J. Belsham; Alejandro Valbuena; Mauricio G. Mateu. Thermostability of the Foot-and-Mouth Disease Virus Capsid Is Modulated by Lethal and Viability-Restoring Compensatory Amino Acid Substitutions. Journal of Virology 2019, 93, 1 .

AMA Style

Silvia López-Argüello, Verónica Rincón, Alicia Rodríguez-Huete, Encarnación Martínez-Salas, Graham J. Belsham, Alejandro Valbuena, Mauricio G. Mateu. Thermostability of the Foot-and-Mouth Disease Virus Capsid Is Modulated by Lethal and Viability-Restoring Compensatory Amino Acid Substitutions. Journal of Virology. 2019; 93 (10):1.

Chicago/Turabian Style

Silvia López-Argüello; Verónica Rincón; Alicia Rodríguez-Huete; Encarnación Martínez-Salas; Graham J. Belsham; Alejandro Valbuena; Mauricio G. Mateu. 2019. "Thermostability of the Foot-and-Mouth Disease Virus Capsid Is Modulated by Lethal and Viability-Restoring Compensatory Amino Acid Substitutions." Journal of Virology 93, no. 10: 1.

Journal article
Published: 13 May 2019 in Journal of Virological Methods
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Foot-and-mouth disease (FMD) remains a globally important disease but there have only been occasional recent outbreaks in Europe, e.g. in the U.K. in 2001, U.K. 2007 and Bulgaria 2010/2011. However, this infection still poses a threat to Europe as the disease occurs close to its borders and incursions can occur through importation of contaminated animal products and through the air. To deal with a suspected outbreak, fast sampling, transportation and accurate laboratory diagnosis are critical; testing for FMDV is normally performed on epithelium samples or serum. Assessment of the use of stabilized blood in assays for FMDV RNA is useful as this sample material can be prepared on site for safe transportation and rapid analysis at the laboratory. Such samples are also collected for diagnosis of other diseases giving similar clinical signs. Testing serum and EDTA-stabilized blood samples from FMDV-infected cattle and pigs, using real time quantitative RT-PCR assays, yielded similar results. However, detection of FMDV RNA was less sensitive (about 10-fold) when using EDTA-stabilized blood compared to serum. Thus, diagnosis of FMD can be achieved using EDTA-stabilized blood samples in an outbreak situation on a herd basis, but serum is preferred at the single animal level for optimal sensitivity.

ACS Style

Kristina S. Fontél; Anette Bøtner; Graham J. Belsham; Louise Lohse. Diagnostic comparison of serum and EDTA-stabilized blood samples for the detection of foot-and-mouth disease virus RNA by RT-qPCR. Journal of Virological Methods 2019, 270, 120 -125.

AMA Style

Kristina S. Fontél, Anette Bøtner, Graham J. Belsham, Louise Lohse. Diagnostic comparison of serum and EDTA-stabilized blood samples for the detection of foot-and-mouth disease virus RNA by RT-qPCR. Journal of Virological Methods. 2019; 270 ():120-125.

Chicago/Turabian Style

Kristina S. Fontél; Anette Bøtner; Graham J. Belsham; Louise Lohse. 2019. "Diagnostic comparison of serum and EDTA-stabilized blood samples for the detection of foot-and-mouth disease virus RNA by RT-qPCR." Journal of Virological Methods 270, no. : 120-125.

Research article
Published: 18 January 2019 in PLOS Pathogens
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Many picornaviruses cause important diseases in humans and other animals including poliovirus, rhinoviruses (causing the common cold) and foot-and-mouth disease virus (FMDV). These small, non-enveloped viruses comprise a positive-stranded RNA genome (ca. 7–9 kb) enclosed within a protein shell composed of 60 copies of three or four different capsid proteins. For the aphthoviruses (e.g. FMDV) and cardioviruses, the capsid precursor, P1-2A, is cleaved by the 3C protease (3Cpro) to generate VP0, VP3 and VP1 plus 2A. For enteroviruses, e.g. poliovirus, the capsid precursor is P1 alone, which is cleaved by the 3CD protease to generate just VP0, VP3 and VP1. The sequences required for correct processing of the FMDV capsid protein precursor in mammalian cells were analyzed. Truncation of the P1-2A precursor from its C-terminus showed that loss of the 2A peptide (18 residues long) and 27 residues from the C-terminus of VP1 (211 residues long) resulted in a precursor that cannot be processed by 3Cpro although it still contained two unmodified internal cleavage sites (VP0/VP3 and VP3/VP1 junctions). Furthermore, introduction of small deletions within P1-2A identified residues 185–190 within VP1 as being required for 3Cpro-mediated processing and for optimal accumulation of the precursor. Within this C-terminal region of VP1, five of these residues (YCPRP), are very highly conserved in all FMDVs and are also conserved amongst other picornaviruses. Mutant FMDV P1-2A precursors with single amino acid substitutions within this motif were highly resistant to cleavage at internal junctions. Such substitutions also abrogated virus infectivity. These results can explain earlier observations that loss of the C-terminus (including the conserved motif) from the poliovirus capsid precursor conferred resistance to processing. Thus, this motif seems essential for maintaining the correct structure of picornavirus capsid precursors prior to processing and subsequent capsid assembly; it may represent a site that interacts with cellular chaperones. The picornavirus family includes clinically important human and animal pathogens, for example: poliovirus, rhinovirus (causing the common cold) and foot-and-mouth disease virus (FMDV) that infects cloven-hoofed animals. Picornaviruses contain a positive-sense RNA genome surrounded by a protein shell, also called a capsid. The capsid proteins are made from a precursor and correct processing and assembly of these capsid proteins is necessary in the virus life cycle to create new infectious virus particles. In this study, we have identified a short motif (just 5 amino acids long) within the capsid precursor, which is highly conserved among picornaviruses. Deletion of this motif inhibited processing of the junctions between the mature structural proteins within this precursor, with one junction being more than 400 amino acids away from this region. This motif also seems to be required for the optimal accumulation of the capsid precursor in cells. We hypothesize that the motif may be involved in binding to a cellular protein, such as a chaperone, to stabilize the capsid precursor and promote its correct folding to allow it to be processed by the viral protease prior to capsid assembly.

ACS Style

Thea Kristensen; Graham J. Belsham. Identification of a short, highly conserved, motif required for picornavirus capsid precursor processing at distal sites. PLOS Pathogens 2019, 15, e1007509 .

AMA Style

Thea Kristensen, Graham J. Belsham. Identification of a short, highly conserved, motif required for picornavirus capsid precursor processing at distal sites. PLOS Pathogens. 2019; 15 (1):e1007509.

Chicago/Turabian Style

Thea Kristensen; Graham J. Belsham. 2019. "Identification of a short, highly conserved, motif required for picornavirus capsid precursor processing at distal sites." PLOS Pathogens 15, no. 1: e1007509.

Journal article
Published: 31 October 2018 in mSphere
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Foot-and-mouth disease (FMD) is a viral disease of livestock with substantial impact on agricultural production and subsistence farming on a global scale. Control of FMD is impeded by the existence of a prolonged asymptomatic carrier phase during which infected cattle shed low quantities of infectious virus in oropharyngeal fluid (OPF) for months to years after infection. The epidemiological significance of FMD virus (FMDV) carriers is unresolved. However, the existence of the FMDV carrier state has substantial impact on international trade in animal products. The current investigation demonstrated that transfer of OPF from persistently infected FMDV carrier cattle to naive cattle led to fulminant clinical FMD. It was thus demonstrated that, although the risk for disease transmission under natural conditions is considered to be low, there is detectable contagion associated with FMDV carrier cattle. This finding is important for optimization of FMD risk mitigation strategies.

ACS Style

Jonathan Arzt; Graham J. Belsham; Louise Lohse; Anette Bøtner; Carolina Stenfeldt. Transmission of Foot-and-Mouth Disease from Persistently Infected Carrier Cattle to Naive Cattle via Transfer of Oropharyngeal Fluid. mSphere 2018, 3, e00365-18 .

AMA Style

Jonathan Arzt, Graham J. Belsham, Louise Lohse, Anette Bøtner, Carolina Stenfeldt. Transmission of Foot-and-Mouth Disease from Persistently Infected Carrier Cattle to Naive Cattle via Transfer of Oropharyngeal Fluid. mSphere. 2018; 3 (5):e00365-18.

Chicago/Turabian Style

Jonathan Arzt; Graham J. Belsham; Louise Lohse; Anette Bøtner; Carolina Stenfeldt. 2018. "Transmission of Foot-and-Mouth Disease from Persistently Infected Carrier Cattle to Naive Cattle via Transfer of Oropharyngeal Fluid." mSphere 3, no. 5: e00365-18.

Journal article
Published: 11 September 2018 in Viruses
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Bat populations harbour a multitude of viruses; some of these are pathogenic or potentially pathogenic in other animals or humans. Therefore, it is important to monitor the populations and characterize these viruses. In this study, the presence of coronaviruses (CoVs) in different species of Danish bats was investigated using active surveillance at different geographical locations in Denmark. Faecal samples were screened for the presence of CoVs using pan-CoV real-time RT-PCR assays. The amplicons, obtained from five different species of bats, were sequenced. Phylogenetic analysis revealed a species-specific clustering with the samples from Myotis daubentonii, showing a close resemblance to coronavirus sequences obtained from the same species of bat in Germany and the United Kingdom. Our results show, for the first time, that multiple, distinct alphacoronaviruses are present in the Danish bat populations.

ACS Style

Christina M. Lazov; Mariann Chriél; Hans J. Baagøe; Esben Fjederholt; Yu Deng; Engbert A. Kooi; Graham J. Belsham; Anette Bøtner; Thomas Bruun Rasmussen. Detection and Characterization of Distinct Alphacoronaviruses in Five Different Bat Species in Denmark. Viruses 2018, 10, 486 .

AMA Style

Christina M. Lazov, Mariann Chriél, Hans J. Baagøe, Esben Fjederholt, Yu Deng, Engbert A. Kooi, Graham J. Belsham, Anette Bøtner, Thomas Bruun Rasmussen. Detection and Characterization of Distinct Alphacoronaviruses in Five Different Bat Species in Denmark. Viruses. 2018; 10 (9):486.

Chicago/Turabian Style

Christina M. Lazov; Mariann Chriél; Hans J. Baagøe; Esben Fjederholt; Yu Deng; Engbert A. Kooi; Graham J. Belsham; Anette Bøtner; Thomas Bruun Rasmussen. 2018. "Detection and Characterization of Distinct Alphacoronaviruses in Five Different Bat Species in Denmark." Viruses 10, no. 9: 486.

Correspondence
Published: 13 August 2018 in Veterinary Microbiology
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ACS Style

Graham J. Belsham; Anette Bøtner. A reply to "A comment on "Inter-laboratory study to characterize the detection of serum antibodies against porcine epidemic diarrhoea virus". Veterinary Microbiology 2018, 224, 118 .

AMA Style

Graham J. Belsham, Anette Bøtner. A reply to "A comment on "Inter-laboratory study to characterize the detection of serum antibodies against porcine epidemic diarrhoea virus". Veterinary Microbiology. 2018; 224 ():118.

Chicago/Turabian Style

Graham J. Belsham; Anette Bøtner. 2018. "A reply to "A comment on "Inter-laboratory study to characterize the detection of serum antibodies against porcine epidemic diarrhoea virus"." Veterinary Microbiology 224, no. : 118.