Publications by authors named "Kris De Clercq"

82 Publications

Transmission of Bluetongue Virus Serotype 8 by Artificial Insemination with Frozen-Thawed Semen from Naturally Infected Bulls.

Viruses 2021 04 9;13(4). Epub 2021 Apr 9.

Unit of Exotic and Particular Diseases, Scientific Directorate Infectious Diseases in Animals, Sciensano, 1180 Brussels, Belgium.

Transmission of bluetongue (BT) virus serotype 8 (BTV-8) via artificial insemination of contaminated frozen semen from naturally infected bulls was investigated in two independent experiments. Healthy, BT negative heifers were hormonally synchronized and artificially inseminated at oestrus. In total, six groups of three heifers received semen from four batches derived from three bulls naturally infected with BTV-8. Each experiment included one control heifer that was not inseminated and that remained BT negative throughout. BTV viraemia and seroconversion were determined in 8 out of 18 inseminated heifers, and BTV was isolated from five of these animals. These eight heifers only displayed mild clinical signs of BT, if any at all, but six of them experienced pregnancy loss between weeks four and eight of gestation, and five of them became BT PCR and antibody positive. The other two infected heifers gave birth at term to two healthy and BT negative calves. The BT viral load varied among the semen batches used and this had a significant impact on the infection rate, the time of onset of viraemia post artificial insemination, and the gestational stage at which pregnancy loss occurred. These results, which confirm unusual features of BTV-8 infection, should not be extrapolated to infection with other BTV strains without thorough evaluation. This study also adds weight to the hypothesis that the re-emergence of BTV-8 in France in 2015 may be attributable to the use of contaminated bovine semen.
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http://dx.doi.org/10.3390/v13040652DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8069090PMC
April 2021

Nearly Complete Genome Sequences of Two Bluetongue Viruses Isolated during the 2020 Outbreak in the Grand Duchy of Luxembourg.

Microbiol Resour Announc 2021 Apr 8;10(14). Epub 2021 Apr 8.

Sciensano, Exotic Viruses and Particular Diseases Unit, Ukkel, Belgium.

Bluetongue is one of the major diseases of ruminants listed by the World Organisation for Animal Health. Bluetongue virus serotype 8 (BTV-8) has been considered enzootic in France since 2018. Here, we report the nearly complete genome sequences of two BTV-8 isolates from the 2020 outbreak in the Grand Duchy of Luxembourg.
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http://dx.doi.org/10.1128/MRA.00210-21DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8032468PMC
April 2021

Scientific Opinion on the assessment of the control measures of the category A diseases of Animal Health Law: .

EFSA J 2021 Feb 3;19(2):e06403. Epub 2021 Feb 3.

EFSA received a mandate from the European Commission to assess the effectiveness of some of the control measures against diseases included in the Category A list according to Regulation (EU) 2016/429 on transmissible animal diseases ('Animal Health Law'). This opinion belongs to a series of opinions where these control measures will be assessed, with this opinion covering the assessment of control measures for African Horse Sickness (AHS). In this opinion, EFSA and the AHAW Panel of experts review the effectiveness of: (i) clinical and laboratory sampling procedures, (ii) monitoring period and (iii) the minimum radius of the protection and surveillance zone, and the minimum duration of measures in these zones. The general methodology used for this series of opinions has been published elsewhere; nonetheless, specific details of the transmission kernels used for the assessment of the minimum radius of the protection and surveillance zones are shown. Several scenarios for which these control measures were assessed were designed and agreed prior to the start of the assessment. In summary, sampling procedures described in the diagnostic manual for AHS were considered efficient for all Equidae considering the high case fatality rate expected. The monitoring period (14 days) was assessed as effective in every scenario, except for those relating to the epidemiological enquiry where the risk manager should consider increasing the monitoring period, based on the awareness of keepers, environmental conditions and the vector abundance in the region. The current protection zone (100 km) comprises more than 95% of the infections from an affected establishment. Both the radius and duration of the zones could be reduced, based on local environmental conditions and the time of year of the first index case. Recommendations provided for each of the scenarios assessed aim to support the European Commission in the drafting of further pieces of legislation relating to AHS.
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http://dx.doi.org/10.2903/j.efsa.2021.6403DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7856565PMC
February 2021

Scientific Opinion on the assessment of the control measures of the category A diseases of Animal Health Law: African Swine Fever.

EFSA J 2021 Jan 31;19(1):e06402. Epub 2021 Jan 31.

EFSA received a mandate from the European Commission to assess the effectiveness of some of the control measures against diseases included in the Category A list according to Regulation (EU) 2016/429 on transmissible animal diseases ('Animal Health Law'). This opinion belongs to a series of opinions where these control measures will be assessed, with this opinion covering the assessment of control measures for African Swine Fever (ASF). In this opinion, EFSA and the AHAW Panel of experts reviewed the effectiveness of: (i) clinical and laboratory sampling procedures, (ii) monitoring period and (iii) the minimum radius of the protection and surveillance zone, and the minimum length of time the measures should be applied in these zones. The general methodology used for this series of opinions has been published elsewhere; nonetheless, specific details of the model used for the assessment of the laboratory sampling procedures for ASF are presented here. Here, also, the transmission kernels used for the assessment of the minimum radius of the protection and surveillance zones are shown. Several scenarios for which these control measures had to be assessed were designed and agreed prior to the start of the assessment. In summary, several sampling procedures as described in the diagnostic manual for ASF were considered ineffective and a suggestion to exclude, or to substitute with more effective procedures was made. The monitoring period was assessed as non-effective for several scenarios and a longer monitoring period was suggested to ensure detection of potentially infected herds. It was demonstrated that the surveillance zone comprises 95% of the infections from an affected establishment, and therefore is considered effective. Recommendations provided for each of the scenarios assessed aim to support the European Commission in the drafting of further pieces of legislation, as well as for plausible ad hoc requests in relation to ASF.
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http://dx.doi.org/10.2903/j.efsa.2021.6402DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7848183PMC
January 2021

Scientific Opinion on the assessment of the control measures of the category A diseases of Animal Health Law: Highly Pathogenic Avian Influenza.

EFSA J 2021 Jan 18;19(1):e06372. Epub 2021 Jan 18.

EFSA received a mandate from the European Commission to assess the effectiveness of some of the control measures against diseases included in the Category A list according to Regulation (EU) 2016/429 on transmissible animal diseases ('Animal Health Law'). This opinion belongs to a series of opinions where these control measures will be assessed, with this opinion covering the assessment of control measures for Highly Pathogenic Avian Influenza (HPAI). In this opinion, EFSA and the AHAW Panel of experts review the effectiveness of: (i) clinical and laboratory sampling procedures, (ii) monitoring period and (iii) the minimum radius of the protection and surveillance zone, and the minimum length of time the measures should be applied in these zones. The general methodology used for this series of opinions has been published elsewhere; nonetheless, specific details of the model used for the assessment of the laboratory sampling procedures for HPAI are presented here. Here, also, the transmission kernels used for the assessment of the minimum radius of the protection and surveillance zones are shown. Several scenarios for which these control measures had to be assessed were designed and agreed prior to the start of the assessment. In summary, sampling procedures as described in the diagnostic manual for HPAI were considered efficient for gallinaceous poultry, whereas additional sampling is advised for Anseriformes. The monitoring period was assessed as effective, and it was demonstrated that the surveillance zone comprises 95% of the infections from an affected establishment. Recommendations provided for each of the scenarios assessed aim to support the European Commission in the drafting of further pieces of legislation, as well as for plausible ad hoc requests in relation to HPAI.
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http://dx.doi.org/10.2903/j.efsa.2021.6372DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7812451PMC
January 2021

The antibody response induced FMDV vaccines in sheep correlates with early transcriptomic responses in blood.

NPJ Vaccines 2020 Jan 3;5(1). Epub 2020 Jan 3.

Université Paris-Saclay, INRA, VIM, Domaine de Vilvert, 78350, Jouy-en-Josas, France.

Foot and mouth disease (FMD) is a highly contagious viral disease with high economic impact, representing a major threat for cloven-hooved mammals worldwide. Vaccines based on adjuvanted inactivated virus (iFMDV) induce effective protective immunity implicating antibody (Ab) responses. To reduce the biosafety constraints of the manufacturing process, a non-replicative human adenovirus type 5 vector encoding FMDV antigens (Ad5-FMDV) has been developed. Here we compared the immunogenicity of iFMDV and Ad5-FMDV with and without the ISA206VG emulsion-type adjuvant in sheep. Contrasted Ab responses were obtained: iFMDV induced the highest Ab levels, Ad5-FMDV the lowest ones, and ISA206VG increased the Ad5-FMDV-induced Ab responses to protective levels. Each vaccine generated heterogeneous Ab responses, with high and low responders, the latter being considered as obstacles to vaccine effectiveness. A transcriptomic study on total blood responses at 24 h post-vaccination revealed several blood gene module activities correlating with long-term Ab responses. Downmodulation of T cell modules' activities correlated with high responses to iFMDV and to Ad5-FMDV+ISA206VG vaccines as also found in other systems vaccinology studies in humans and sheep. The impact of cell cycle activity depended on the vaccine types, as it positively correlated with higher responses to iFMDV but negatively to non-adjuvanted Ad5-FMDV. Finally an elevated B cell activity at 24 h correlated with high Ab responses to the Ad5-FMDV+ISA206VG vaccine. This study provides insights into the early mechanisms driving the Ab response induced by different vaccine regimens including Ad5 vectors and points to T cell modules as early biomarker candidates of different vaccine-type efficacy across species.
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http://dx.doi.org/10.1038/s41541-019-0151-3DOI Listing
January 2020

Non-discriminatory Exclusion Testing as a Tool for the Early Detection of Foot-and-Mouth Disease Incursions.

Front Vet Sci 2020 19;7:552670. Epub 2020 Nov 19.

Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.

Endemic circulation of foot-and-mouth disease (FMD) in Africa and Asia poses a continuous risk to countries in Europe, North America, and Oceania which are free from the disease. Introductions of the disease into a free region have dramatic economic impacts, especially if they are not detected at an early stage and controlled rapidly. However, farmers and veterinarians have an obvious disincentive to report clinical signs that are consistent with FMD, due to the severe consequences of raising an official suspicion, such as farm-level quarantine. One way that the risk of late detection can be mitigated is offering non-discriminatory exclusion testing schemes for differential diagnostics, wherein veterinarians can submit samples without the involvement of the competent authority and without sanctions or costs for the farmer. This review considers the benefits and limitations of this approach to improve the early detection of FMD in free countries and gives an overview of the FMD testing schemes currently in use in selected countries in Europe and the Americas as well as in Australia.
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http://dx.doi.org/10.3389/fvets.2020.552670DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7710516PMC
November 2020

Complete Coding Sequence of a Lumpy Skin Disease Virus from an Outbreak in Bulgaria in 2016.

Microbiol Resour Announc 2020 Oct 22;9(43). Epub 2020 Oct 22.

Sciensano, EURL Capripox, Unit Exotic Viruses and Particular Diseases, Ukkel, Belgium.

Lumpy skin disease (LSD) is an emerging cattle disease with serious economic consequences. We report the complete coding sequence of LSD virus 210LSD-249/BUL/16, detected in a blood sample from a diseased cow during an outbreak in Bulgaria (Kabile Village, Yambol Region) in June 2016.
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http://dx.doi.org/10.1128/MRA.00977-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7585842PMC
October 2020

Complex Circulation of Foot-and-Mouth Disease Virus in Cattle in Nigeria.

Front Vet Sci 2020 20;7:466. Epub 2020 Aug 20.

Sciensano, Scientific Direction of Infectious Diseases in Animals, Service for Exotic Viruses and Particular Diseases, Brussels, Belgium.

Nigeria is a large densely populated country in West Africa. Most of its livestock is raised in a pastoralist production system with typical long distance migration in search of water and feed. As the demand for animal products largely exceeds the domestic production, large numbers of livestock are imported from neighboring countries without sanitary restrictions. In Nigeria, foot-and-mouth disease virus (FMDV) serotypes O, A, and Southern African Territories (SAT)2 are endemic for a long time. Clinical outbreaks of FMD due to serotype SAT1 are described again since 2015, after an absence of more than 30 years. Historically, outbreaks of FMD due to serotypes O, A, SAT1, and SAT2 were each time associated with trade of cattle entering Nigeria from neighboring countries. In the present study, tissue samples from 27 outbreaks of FMD were collected in Nigerian cattle from 2012 until 2017 in six different States and in the Federal Capital Territory. FMDV was isolated and serotyped and further characterized by VP1 sequencing and phylogenetic analysis to gain more knowledge on FMDV circulation in Nigeria. Half of the outbreaks were characterized as FMDV topotype O/EA-3, while outbreaks with other serotypes and topotypes were-in descending order-less prevalent: A/Africa/G-IV, SAT1/X, SAT2/VII, and O/WA. The high dynamics and omnipresence of FMD in Nigeria were illustrated in Plateau State where FMDV serotypes O, SAT1, and SAT2 were isolated during the course of the study, while at some point in the study, outbreaks due to FMDV serotype A were observed in three remote States. The genetic and phylogenetic analysis suggests a mixed origin of FMD outbreaks. Some outbreaks seem to be caused by sustained local transmission of FMDV strains present in Nigeria since a number of years, while other outbreaks seem to be related to recent incursions with new FMDV strains. The role of African buffaloes in the etiology of FMD in Nigeria is unclear, and sampling of wildlife is needed. The results of the present study suggest that systematic sample collection is essential to understand the complex concomitance of FMDV strains in Nigeria and essential to support the implementation of a vaccination-based control plan.
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http://dx.doi.org/10.3389/fvets.2020.00466DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7468398PMC
August 2020

Lumpy skin disease epidemiological report IV: data collection and analysis.

EFSA J 2020 Feb 27;18(2):e06010. Epub 2020 Feb 27.

In 2019, no lumpy skin disease (LSD) outbreaks were reported in South-Eastern Europe, the mass vaccination regional campaign with homologous LSD vaccine continued for the fourth year with over 1.8 million bovines vaccinated in the region, preventing further outbreaks since 2016. LSD outbreaks were reported in Turkey, including western Turkey, in Russia and in eastern Asia affecting China, Bangladesh and India for the first time. The use of homologous vaccine should be considered in the countries still affected in order to eliminate the virus. Besides passive surveillance, which is implemented in all the countries, active surveillance for early detection based on clinical examination could be conducted ideally during April-October every 5 weeks in at-risk areas, based on possible re-emergence or re-introduction from affected neighbouring countries. Active surveillance for proving disease freedom could be based on serological testing (enzyme-linked immunosorbent assay (ELISA)) targeting 3.5% seroprevalence and conducted on a random sample of cattle herds on non-vaccinated animals. LSD re-emerged in Israel in 2019, after vaccination became voluntary. This shows that, if the virus is still circulating in the region, the reduced protection might result in re-emergence of LSD. In case of re-emergence, a contingency plan and vaccine stockpiling would be needed, in order to react quickly. From a study performed in Israel to test side effects of live-attenuated homologous LSD vaccine, milk production can be reduced during 7 days after vaccination (around 6-8 kg per cow), without a significant loss in the 30 days after vaccination. Research needs should be focused on the probability of transmission from insect to bovine, the virus inactivation rate in insects, the collection of baseline entomological data, the capacity of vector species in LSDV transmission linked to studies on their abundance and the control of being the most important vector in LSD transmission.
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http://dx.doi.org/10.2903/j.efsa.2020.6010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7448019PMC
February 2020

Improving laboratory diagnostic capacities of emerging diseases using knowledge mapping.

Transbound Emerg Dis 2020 Aug 4. Epub 2020 Aug 4.

Research Unit in Epidemiology and Risk Analysis applied to veterinary sciences (UREAR-ULiège), Faculty of Veterinary Medicine, Fundamental and Applied Research for Animal and Health (FARAH) Center, Liège, Belgium.

Over the last decade, European countries faced several emerging and re-emerging animal diseases as well as zoonotic diseases. During these episodes, the laboratory diagnostic capabilities were a key factor to rapidly control and/or eradicate them. Because of the associated socio-economic and health consequences, it is crucial to react rapidly and efficiently, not only during crisis but also in peacetime (i.e. preparedness). However, to date, there is no published method to identify diseases with diagnostic gaps and to prioritize assays to be implemented. This study was conducted based on the outcome of a prioritization exercise in which 29 epizootic and exotic diseases with high risk of emergence or re-emergence in Belgium (Bianchini et al., [2020] Transboundary and Emerging Diseases, 67(1), 344-376) were listed. Knowledge mapping was used to visualize and identify gaps in the diagnostic procedures for different epidemiological scenarios at national level. To fill these gaps, an overview of diagnostic capabilities at national and international level (laboratories and kits providers or manufacturers) as well as the published assays in the scientific literature and the prescribed assays by international institutions and kits providers was carried out. The outcome of this study revealed the usefulness of knowledge mapping as a tool to identify gaps and ultimately gain insight on alternatives for better preparedness and responsiveness. While this exercise was limited to Belgium, we believe this exercise can benefit other countries and thereby enhancing knowledge sharing and collaboration to increase diagnostic capabilities for a common list of (re-) emerging diseases in crisis situation.
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http://dx.doi.org/10.1111/tbed.13768DOI Listing
August 2020

"Frozen evolution" of an RNA virus suggests accidental release as a potential cause of arbovirus re-emergence.

PLoS Biol 2020 04 28;18(4):e3000673. Epub 2020 Apr 28.

MRC-University of Glasgow Centre for Virus Research, Glasgow, United Kingdom.

The mechanisms underlying virus emergence are rarely well understood, making the appearance of outbreaks largely unpredictable. Bluetongue virus serotype 8 (BTV-8), an arthropod-borne virus of ruminants, emerged in livestock in northern Europe in 2006, spreading to most European countries by 2009 and causing losses of billions of euros. Although the outbreak was successfully controlled through vaccination by early 2010, puzzlingly, a closely related BTV-8 strain re-emerged in France in 2015, triggering a second outbreak that is still ongoing. The origin of this virus and the mechanisms underlying its re-emergence are unknown. Here, we performed phylogenetic analyses of 164 whole BTV-8 genomes sampled throughout the two outbreaks. We demonstrate consistent clock-like virus evolution during both epizootics but found negligible evolutionary change between them. We estimate that the ancestor of the second outbreak dates from the height of the first outbreak in 2008. This implies that the virus had not been replicating for multiple years prior to its re-emergence in 2015. Given the absence of any known natural mechanism that could explain BTV-8 persistence over this long period without replication, we hypothesise that the second outbreak could have been initiated by accidental exposure of livestock to frozen material contaminated with virus from approximately 2008. Our work highlights new targets for pathogen surveillance programmes in livestock and illustrates the power of genomic epidemiology to identify pathways of infectious disease emergence.
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http://dx.doi.org/10.1371/journal.pbio.3000673DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7188197PMC
April 2020

Lumpy Skin Disease Is Characterized by Severe Multifocal Dermatitis With Necrotizing Fibrinoid Vasculitis Following Experimental Infection.

Vet Pathol 2020 05;57(3):388-396

The Pirbright Institute, Surrey, UK.

Lumpy skin disease is a high-consequence disease in cattle caused by infection with the poxvirus lumpy skin disease virus (LSDV). The virus is endemic in most countries in Africa and an emerging threat to cattle populations in Europe and Asia. As LSDV spreads into new regions, it is important that signs of disease are recognized promptly by animal caregivers. This study describes the gross, microscopic, and ultrastructural changes that occur over time in cattle experimentally challenged with LSDV. Four calves were inoculated with wildtype LSDV and monitored for 19 to 21 days. At 7 days after inoculation, 2 of the 4 cattle developed multifocal cutaneous nodules characteristic of LSD. Some lesions displayed a targetoid appearance. Histologically, intercellular and intracellular edema was present in the epidermis of some nodules. Occasional intracytoplasmic inclusion bodies were identified in keratinocytes. More severe and consistent changes were present in the dermis, with marked histiocytic inflammation and necrotizing fibrinoid vasculitis of dermal vessels, particularly the deep dermal plexus. Chronic lesions consisted of full-thickness necrosis of the dermis and epidermis. Lesions in other body organs were not a major feature of LSD in this study, highlighting the strong cutaneous tropism of this virus. Immunohistochemistry and electron microscopy identified LSDV-infected histiocytes and fibroblasts in the skin nodules of affected cattle. This study highlights the noteworthy lesions of LSDV and how they develop over time.
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http://dx.doi.org/10.1177/0300985820913268DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7201124PMC
May 2020

Complete Coding Sequence of a Lumpy Skin Disease Virus Strain Isolated during the 2016 Outbreak in Kazakhstan.

Microbiol Resour Announc 2020 Jan 23;9(4). Epub 2020 Jan 23.

Sciensano, Unit Exotic Viruses and Particular Diseases, Ukkel, Belgium.

Lumpy skin disease virus (LSDV) causes an economically important disease in cattle. Here, we report the complete coding sequence of the LSDV isolate Kubash/KAZ/16, detected in a clinical sample from an infected cow from the outbreak reported on 7 July 2016 in Kazakhstan (Atyrau Region).
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http://dx.doi.org/10.1128/MRA.01399-19DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6979303PMC
January 2020

The antibody response induced FMDV vaccines in sheep correlates with early transcriptomic responses in blood.

NPJ Vaccines 2020 3;5. Epub 2020 Jan 3.

Université Paris-Saclay, INRA, VIM, Domaine de Vilvert, 78350 Jouy-en-Josas, France.

Foot and mouth disease (FMD) is a highly contagious viral disease with high economic impact, representing a major threat for cloven-hooved mammals worldwide. Vaccines based on adjuvanted inactivated virus (iFMDV) induce effective protective immunity implicating antibody (Ab) responses. To reduce the biosafety constraints of the manufacturing process, a non-replicative human adenovirus type 5 vector encoding FMDV antigens (Ad5-FMDV) has been developed. Here we compared the immunogenicity of iFMDV and Ad5-FMDV with and without the ISA206VG emulsion-type adjuvant in sheep. Contrasted Ab responses were obtained: iFMDV induced the highest Ab levels, Ad5-FMDV the lowest ones, and ISA206VG increased the Ad5-FMDV-induced Ab responses to protective levels. Each vaccine generated heterogeneous Ab responses, with high and low responders, the latter being considered as obstacles to vaccine effectiveness. A transcriptomic study on total blood responses at 24 h post-vaccination revealed several blood gene module activities correlating with long-term Ab responses. Downmodulation of T cell modules' activities correlated with high responses to iFMDV and to Ad5-FMDV+ISA206VG vaccines as also found in other systems vaccinology studies in humans and sheep. The impact of cell cycle activity depended on the vaccine types, as it positively correlated with higher responses to iFMDV but negatively to non-adjuvanted Ad5-FMDV. Finally an elevated B cell activity at 24 h correlated with high Ab responses to the Ad5-FMDV+ISA206VG vaccine. This study provides insights into the early mechanisms driving the Ab response induced by different vaccine regimens including Ad5 vectors and points to T cell modules as early biomarker candidates of different vaccine-type efficacy across species.
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http://dx.doi.org/10.1038/s41541-019-0151-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6941976PMC
January 2020

Failure to Remove Bluetongue Serotype 8 Virus (BTV-8) From Produced and Derived Bovine Embryos and Subsequent Transmission of BTV-8 to Recipient Cows After Embryo Transfer.

Front Vet Sci 2019 5;6:432. Epub 2019 Dec 5.

Unit of Exotic and Particular Diseases, Sciensano, Brussels, Belgium.

The behavior of BTV-8 in cattle is different from most other serotypes not only with regards to clinical signs but certainly with respect to virus transmission (transplacental, contact). Therefore, the possibility of virus transmission by means of embryo transfer was examined by exposure of produced and derived bovine blastocysts to BTV-8 followed by different washing protocols, including longer exposure times (up to 120 s) to 0.25% trypsin at room temperature or at 37°C. None of the washing protocols used was successful in removing the viral genome completely from the produced and derived embryos as was demonstrated by real-time PCR. Moreover, BTV-8 virus was transmitted to recipient cows after embryo transfer of derived BTV8-exposed embryos, which had been subjected to routine decontamination as recommended by IETS, consisting of 5 washes in PBS followed by a double treatment of 0.25% trypsin for 45s at 37°C, and an additional 5 washes in PBS with 2% FCS. This study clearly demonstrates the necessity of vigorous application of the directives for screening of potential donors and the collected embryos, especially in regions with BTV-8, to prevent transmission of the disease.
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http://dx.doi.org/10.3389/fvets.2019.00432DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6907088PMC
December 2019

An Immunoperoxidase Monolayer Assay (IPMA) for the detection of lumpy skin disease antibodies.

J Virol Methods 2020 03 16;277:113800. Epub 2019 Dec 16.

Sciensano, Infectious Diseases in Animals, Exotic and Particular Diseases, Groeselenberg 99, B-1180, Brussels, Belgium. Electronic address:

During this study a new Immunoperoxidase Monolayer Assay (IPMA) was developed for the detection of antibodies against lumpy skin disease virus (LSDV) in an easy and low tech setting. Using two dilutions (1:50 and 1:300) in a duplicate format, the test was shown to be highly sensitive, specific and repeatable. In comparison to the VNT and a commercial ELISA, the LSDV-IPMA was able to detect the LSDV antibodies earlier in infected, vaccinated and vaccinated/infected animals. The assay is very flexible as it can be easily adapted for the detection of sheeppox or goatpox antibodies and it can be scaled-up to handle medium size sample sets by preparing the IPMA plates in advance. These plates are safe and can be handled in low biosafety level labs.
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http://dx.doi.org/10.1016/j.jviromet.2019.113800DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6996284PMC
March 2020

FMD vaccine matching: Inter laboratory study for improved understanding of r values.

J Virol Methods 2020 02 22;276:113786. Epub 2019 Nov 22.

Unit Exotic Viruses and Particular Diseases, SD Infectious Diseases in Animals, Sciensano (formerly CODA-CERVA), Groeselenberg 99, 1180 Brussel, Belgium. Electronic address:

Foot-and-mouth disease virus (FMDV) is a highly variable RNA virus existing as seven different serotypes. The antigenic variability between and within serotypes can limit the cross-reactivity and therefore the in vivo cross-protection of vaccines. Selection of appropriate vaccine strains is crucial in the control of FMD. Determination of indirect relationships (r-value) between potential vaccine strains and field strains based on antibody responses against both are routinely used for vaccine matching purposes. Aiming at the investigation of the repeatability, reproducibility and comparability of r-value determination within and between laboratories and serological tests, a small scale vaccine matching ring test for FMDV serotype A was organized. Well-characterized serum pools from cattle vaccinated with a monovalent A24/Cruzeiro/Brazil/55 (A24) FMD vaccine with known in vivo protection status (homologous and heterologous) were distributed to four laboratories to determine r-values for the heterologous FMD strains A81/Argentina/87, A/Argentina/2000 and A/Argentina/2001 using the virus neutralization tests (VNT) and liquid phase blocking ELISA (LPBE). Within laboratories, the repeatability of r-value determination was high for both antibody assays. VNT resulted in reproducible and comparable r-values between laboratories, indicative of a lack of antigenic relatedness between the A24 strain and the heterologous strains tested in this work, thus corresponding to some of the in vivo findings with these strains. Using LPBE, similar trends in r-values were observed in all laboratories, but the overall reproducibility was lower than with VNT. Inconsistencies between laboratories may at least in part be attributed to differences in LPBE protocols as well as the in preexisting information generated in each laboratory (such as antibody titer-protection correlation curves). To gain more insight in the LPBE-derived r-values standard bovine control sera were included in the antibody assays performed in each laboratory and a standardization exercise was performed.
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http://dx.doi.org/10.1016/j.jviromet.2019.113786DOI Listing
February 2020

Prioritization of livestock transboundary diseases in Belgium using a multicriteria decision analysis tool based on drivers of emergence.

Transbound Emerg Dis 2020 Jan 9;67(1):344-376. Epub 2019 Oct 9.

Faculty of Veterinary Medicine, Research Unit in Epidemiology and Risk Analysis Applied to Veterinary Sciences (UREAR-ULiege), Fundamental and Applied Research for Animals & Health (FARAH), Centre, Liege University, Liege, Belgium.

During the past decade, livestock diseases have (re-)emerged in areas where they had been previously eradicated or never been recorded before. Drivers (i.e. factors of (re-)emergence) have been identified. Livestock diseases spread irrespective of borders, and therefore, reliable methods are required to help decision-makers to identify potential threats and try stopping their (re-)emergence. Ranking methods and multicriteria approaches are cost-effective tools for such purpose and were applied to prioritize a list of selected diseases (N = 29 including 6 zoonoses) based on the opinion of 62 experts in accordance with 50 drivers-related criteria. Diseases appearing in the upper ranking were porcine epidemic diarrhoea, foot-and-mouth disease, low pathogenic avian influenza, African horse sickness and highly pathogenic avian influenza. The tool proposed uses a multicriteria decision analysis approach to prioritize pathogens according to drivers and can be applied to other countries or diseases.
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http://dx.doi.org/10.1111/tbed.13356DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7168563PMC
January 2020

Reliable and Standardized Animal Models to Study the Pathogenesis of Bluetongue and Schmallenberg Viruses in Ruminant Natural Host Species with Special Emphasis on Placental Crossing.

Viruses 2019 08 15;11(8). Epub 2019 Aug 15.

Research Unit in Epidemiology and Risk Analysis Applied to Veterinary Sciences (UREAR-ULg), Fundamental and Applied Research for Animals & Health (FARAH) Center, Faculty of Veterinary Medicine, University of Liege, 4000 Liege, Belgium.

Starting in 2006, bluetongue virus serotype 8 (BTV8) was responsible for a major epizootic in Western and Northern Europe. The magnitude and spread of the disease were surprisingly high and the control of BTV improved significantly with the marketing of BTV8 inactivated vaccines in 2008. During late summer of 2011, a first cluster of reduced milk yield, fever, and diarrhoea was reported in the Netherlands. Congenital malformations appeared in March 2012 and Schmallenberg virus (SBV) was identified, becoming one of the very few orthobunyaviruses distributed in Europe. At the start of both epizootics, little was known about the pathogenesis and epidemiology of these viruses in the European context and most assumptions were extrapolated based on other related viruses and/or other regions of the World. Standardized and repeatable models potentially mimicking clinical signs observed in the field are required to study the pathogenesis of these infections, and to clarify their ability to cross the placental barrier. This review presents some of the latest experimental designs for infectious disease challenges with BTV or SBV. Infectious doses, routes of infection, inoculum preparation, and origin are discussed. Particular emphasis is given to the placental crossing associated with these two viruses.
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http://dx.doi.org/10.3390/v11080753DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6722754PMC
August 2019

Overview of diagnostic tools for Capripox virus infections.

Prev Vet Med 2020 Aug 28;181:104704. Epub 2019 May 28.

Sciensano, Exotic and Particular Diseases, Groeselenberg 99, 1180, Ukkel, Belgium. Electronic address:

Capripox viruses are the causative agents of important animal diseases in cattle (Lumpy Skin Disease), sheep (Sheeppox) and goats (Goatpox) with severe socio-economic impact in case of wide scale outbreaks. Therefore there is a constant need for adequate diagnostic tools. The assays must be fit-for-purpose to identify the virus quickly and correctly and to be useful for surveillance and monitoring at different stages of an epidemic. Different diagnostic performance characteristics are required depending on the situation and the test purpose. The need for high throughput, high specificity/sensitivity and the capability for differentiating field virus strains from vaccine strains drives the development of new and better assays preferably with an advantageous cost-benefit balance. This review aims to look at existing and new virological and serological diagnostic tools used in the control against diseases caused by Capripox viruses.
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http://dx.doi.org/10.1016/j.prevetmed.2019.104704DOI Listing
August 2020

Serological and molecular epidemiology of foot-and-mouth disease viruses in agro-pastoralist livestock herds in the kachia grazing reserve, Nigeria.

Transbound Emerg Dis 2019 Jul 15;66(4):1575-1586. Epub 2019 May 15.

Vesicular and Exotic Diseases Unit, Veterinary and Agrochemical Research Centre (CODA-CERVA), Brussels, Belgium.

The Kachia Grazing Reserve (KGR) is located in Kaduna state in north-western Nigeria and consists of 6 contiguous blocks housing 744 defined households (HH), all engaged in livestock keeping. It is considered as a homogenous epidemiological unit and a defined study area. In 2012, all cattle and sheep of 40 selected HH were sampled to determine sero-prevalence of antibodies to foot-and-mouth disease virus (FMDV) and of FMDV. The overall sero-prevalence of antibodies to the non-structural 3ABC protein (NSP-3ABC ELISA) was 28.9% (380/1,315) (30.6% cattle; 16.3% sheep), and in 4.5% (62/1,380) (5% cattle; 0.6% sheep) of the examined sera FMD viral RNA could be detected by real-time RT-PCR (rRT-PCR). Additionally, in 2012 and 2014 serum, epithelium and probang samples were collected from cattle in reported FMD outbreaks and the causative FMDVs were molecularly characterized. Approximately half (28/59) of the outbreak sera reacted positive in NSP-3ABC ELISA, and 88% (52/59) of the outbreak sera contained detectable viral RNA. Overall, antibodies against five FMDV serotypes (O, A, SAT1, SAT2 and SAT3) were detected by solid phase competitive ELISA with combinations of two or more serotypes being common. Of the 21 FMDVs that could be isolated 19 were sequenced and 18 were confirmed as SAT2 (lineage VII) while one was characterized as serotype O (EA-3 topotype). Phylogenetic analysis revealed a close relationship between Nigerian FMDV strains and strains in this region and even with strains in North-Africa. Our findings indicate that FMD constitutes an endemic health problem to cattle rearing in the agro-pastoralist community in the KGR and that the KGR is not a closed epidemiological unit. Insight into the local FMDV epidemiology and in the circulating FMDV serotypes/strains is of support to the relevant authorities in Nigeria when considering the need for an FMD control policy to improve animal production in grazing reserves.
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http://dx.doi.org/10.1111/tbed.13182DOI Listing
July 2019

A simple method to estimate the number of doses to include in a bank of vaccines. The case of Lumpy Skin Disease in France.

PLoS One 2019 25;14(1):e0210317. Epub 2019 Jan 25.

IRTA, Centre de Recerca en Sanitat Animal (CReSA, IRTA-UAB), Bellaterra, Barcelona, Spain.

A simple method to estimate the size of the vaccine bank needed to control an epidemic of an exotic infectious disease in case of introduction into a country is presented. The method was applied to the case of a Lumpy Skin disease (LSD) epidemic in France. The size of the stock of vaccines needed was calculated based on a series of simple equations that use some trigonometric functions and take into account the spread of the disease, the time required to obtain good vaccination coverage and the cattle density in the affected region. Assuming a 7-weeks period to vaccinate all the animals and a spread of the disease of 7.3 km/week, the vaccination of 740 716 cattle would be enough to control an epidemic of LSD in France in 90% of the simulations (608 196 cattle would cover 75% of the simulations). The results of this simple method were then validated using a dynamic simulation model, which served as reference for the calculation of the vaccine stock required. The differences between both models in different scenarios, related with the time needed to vaccinate the animals, ranged from 7% to 10.5% more vaccines using the simple method to cover 90% of the simulations, and from 9.0% to 13.8% for 75% of the simulations. The model is easy to use and may be adapted for the control of different diseases in different countries, just by using some simple formulas and few input data.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0210317PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6347152PMC
October 2019

Risk of introduction of Lumpy Skin Disease into France through imports of cattle.

Transbound Emerg Dis 2019 Mar 5;66(2):957-967. Epub 2019 Feb 5.

Members of the Expert Committee for Animal Health and Welfare, ANSES, Maisons-Alfort, France.

The lumpy skin disease (LSD) virus belongs to the genus Capripoxvirus and causes a disease in cattle with economic impacts. In November 2014, the disease was first reported in Europe (in Cyprus); it was then reported in Greece (in August 2015) and has spread through different Balkan countries since 2016. Although vector transmission is predominant in at-risk areas, long-distance transmission usually occurs through movements of infected cattle. In order to estimate the threat for France, a quantitative import risk analysis (QIRA) model was developed to assess the risk of LSD being introduced into France by imports of cattle. Based on available information and using a stochastic model, the probability of a first outbreak of LSD in France following the import of batches of infected live cattle for breeding or fattening was estimated to be 5.4 × 10 (95% probability interval [PI]: 0.4 × 10 ; 28.7 × 10 ) in summer months (during high vector activity) and 1.8 × 10 (95% PI: 0.14 × 10 ; 15 × 10 ) in winter months. The development of a stochastic QIRA made it possible to quantify the risk of LSD being introduced into France through imports of live cattle. This tool is of prime importance because the LSD situation in the Balkans is continuously changing. Indeed, this model can be updated to process new information on the changing health situation in addition to new data from the TRAde Control and Expert System (TRACES, EU database). This model is easy to adapt to different countries and to other diseases.
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http://dx.doi.org/10.1111/tbed.13111DOI Listing
March 2019

Review of epidemiological risk models for foot-and-mouth disease: Implications for prevention strategies with a focus on Africa.

PLoS One 2018 13;13(12):e0208296. Epub 2018 Dec 13.

Research Unit in Epidemiology and Risk Analysis Applied to Veterinary Sciences (UREAR-ULiège), Fundamental and Applied Research for Animals & Health (FARAH) Centre, Faculty of Veterinary Medicine, University of Liege, Liege, Belgium.

Foot-and-mouth disease (FMD) is a highly infectious transboundary disease that affects domestic and wild cloven-hoofed animal species. The aim of this review was to identify and critically assess some modelling techniques for FMD that are well supported by scientific evidence from the literature with a focus on their use in African countries where the disease remains enzootic. In particular, this study attempted to provide a synopsis of the relative strengths and weaknesses of these models and their relevance to FMD prevention policies. A literature search was conducted to identify quantitative and qualitative risk assessments for FMD, including studies that describe FMD risk factor modelling and spatiotemporal analysis. A description of retrieved papers and a critical assessment of the modelling methods, main findings and their limitations were performed. Different types of models have been used depending on the purpose of the study and the nature of available data. The most frequently identified factors associated with the risk of FMD occurrence were the movement (especially uncontrolled animal movement) and the mixing of animals around water and grazing points. Based on the qualitative and quantitative risk assessment studies, the critical pathway analysis showed that the overall risk of FMDV entering a given country is low. However, in some cases, this risk can be elevated, especially when illegal importation of meat and the movement of terrestrial livestock are involved. Depending on the approach used, these studies highlight shortcomings associated with the application of models and the lack of reliable data from endemic settings. Therefore, the development and application of specific models for use in FMD endemic countries including Africa is encouraged.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0208296PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6292601PMC
May 2019

Effectiveness and cost-benefit study to encourage herd owners in a cost sharing vaccination programme against bluetongue serotype-8 in Belgium.

Transbound Emerg Dis 2019 Jan 26;66(1):400-411. Epub 2018 Oct 26.

Epidemiology and Public Health, Veterinary Epidemiology, Brussels, Belgium.

Bluetongue (BT) is a ruminant viral infectious disease transmitted by Culicoides spp. midges. In 2006, when bluetongue virus serotype 8 (BTV-8) appeared for the first time in Northern Europe, it rapidly spread and infected a large proportion of animals. BThas a significant economic impact due to a direct effect on animal health and to an indirect effect in disrupting international trade of animals and animal products. In spring 2008, a compulsory subsidized vaccination programme in Europe resulted in a drastic decrease in the number of reported cases. However, due to the turn-over of the population, without a continuous vaccination programme, the animal population was becoming progressively susceptible. Vaccination would enable Belgium to maintain its status of freedom from infection of BTV-8 that could possibly be re-introduced. Subsidizing it could be an incentive to convince more farmers to vaccinate. To finance this programme, both decision-makers and stakeholders need to be persuaded by the effectiveness and the cost-benefit of vaccination. The study evaluated the effectiveness of vaccination against BTV-8 in Belgium. The change in serology which has shown the effectiveness of the vaccine to induce antibody production has been significantly associated with the time between the first injection and the sampling date and the number of injections of the primo-vaccination. This study also clearly confirms the benefit of vaccination by reducing economic impact of treatment and production losses, especially in dairy cattle. Based on a participating epidemiological approach, a national voluntary and subsidized vaccination was accepted, and permitted Belgium to vaccinate more than 9,000 herds in 1 month. Because this mass vaccination occurred before the vector season, it probably helped Belgium remain free from BTV-8.
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http://dx.doi.org/10.1111/tbed.13034DOI Listing
January 2019

Assessment of cross-protection induced by a bluetongue virus (BTV) serotype 8 vaccine towards other BTV serotypes in experimental conditions.

Vet Res 2018 07 16;49(1):63. Epub 2018 Jul 16.

Faculty of Veterinary Medicine, Research Unit in Epidemiology and Risk Analysis Applied to Veterinary Sciences (UREAR-ULg), Fundamental and Applied Research for Animal and Health (FARAH) Center, University of Liege, Liege, Belgium.

Bluetongue disease is caused by bluetongue virus (BTV) and BTV serotype 8 (BTV8) caused great economic damage in Europe during the last decade. From 1998 to 2007, in addition to BTV8, Europe had to face the emergence of BTV1, 2, 4, 9, and 16, spreading in countries where the virus has never been detected before. These unprecedented outbreaks trigger the need to evaluate and compare the clinical, virological and serological features of the European BTV serotypes in the local epidemiological context. In this study groups of calves were infected with one of the following European BTV serotypes, namely BTV1, 2, 4, 9 and 16. For each tested serotype, two groups of three male Holstein calves were used: one group vaccinated against BTV8, the other non-vaccinated. Clinical signs were quantified, viral RNA was detected in blood and organs and serological relationship was assessed. Calves were euthanized 35 days post-infection and necropsied. Most of the infected animals showed mild clinical signs. A partial serological cross reactivity has been reported between BTV8 and BTV4, and between BTV1 and BTV8. BTV2 and BTV4 viral RNA only reached low levels in blood, when compared to other serotypes, whereas in vitro growth assays could not highlight significant differences. Altogether the results of this study support the hypothesis of higher adaptation of some BTV strains to specific hosts, in this case calves. Furthermore, cross-protection resulting from a prior vaccination with BTV8 was highlighted based on cross-neutralization. However, the development of neutralizing antibodies is probably not totally explaining the mild protection induced by the heterologous vaccination.
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http://dx.doi.org/10.1186/s13567-018-0556-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6048908PMC
July 2018

Risk of introduction of lumpy skin disease in France by the import of vectors in animal trucks.

PLoS One 2018 11;13(6):e0198506. Epub 2018 Jun 11.

Members of the Expert Committee for Animal Health and Welfare, ANSES, Maisons-Alfort, France.

Background: The lumpy skin disease virus (LSDV) is a dsDNA virus belonging to the Poxviridae family and the Capripoxvirus genus. Lumpy skin diseases (LSD) is a highly contagious transboundary disease in cattle producing major economic losses. In 2014, the disease was first reported in the European Union (in Cyprus); it was then reported in 2015 (in Greece) and has spread through different Balkan countries in 2016. Indirect vector transmission is predominant at small distances, but transmission between distant herds and between countries usually occurs through movements of infected cattle or through vectors found mainly in animal trucks.

Methods And Principal Findings: In order to estimate the threat for France due to the introduction of vectors found in animal trucks (cattle or horses) from at-risk countries (Balkans and neighbours), a quantitative import risk analysis (QIRA) model was developed according to the international standard. Using stochastic QIRA modelling and combining experimental/field data and expert opinion, the yearly risk of LSDV being introduced by stable flies (Stomoxys calcitrans), that travel in trucks transporting animals was between 6 x 10-5 and 5.93 x 10-3 with a median value of 89.9 x 10-5; it was mainly due to the risk related to insects entering farms in France from vehicles transporting cattle from the at-risk area. The risk related to the transport of cattle going to slaughterhouses or the transport of horses was much lower (between 2 x 10-7 and 3.73 x 10-5 and between 5 x 10-10 and 3.95 x 10-8 for cattle and horses, respectively). The disinsectisation of trucks transporting live animals was important to reduce this risk.

Conclusion And Significance: The development of a stochastic QIRA made it possible to quantify the risk of LSD being introduced in France through the import of vectors that travel in trucks transporting animals. This tool is of prime importance because the LSD situation in the Balkans is continuously changing. Indeed, this model can be updated to process new information on vectors and the changing health situation, in addition to new data from the TRAde Control and Expert System (TRACES, EU database). This model is easy to adapt to different countries and to other vectors and diseases.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0198506PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5995388PMC
December 2018

A canine adenovirus type 2 vaccine vector confers protection against foot-and-mouth disease in guinea pigs.

Vaccine 2018 04 12;36(16):2193-2198. Epub 2018 Mar 12.

UMR Virologie, INRA, ANSES, École Nationale Vétérinaire d'Alfort, Maisons-Alfort, F-94700, France. Electronic address:

Vaccination is a key element in the control of foot-and-mouth disease (FMD). The majority of the antigenic sites that induce protective immune responses are localized on the FMD virus (FMDV) capsid that is formed by four virus-encoded structural proteins, VP1 to VP4. In the present study, recombinant canine adenovirus type 2 (CAV2)-based FMD vaccines, Cav-P1/3C R° and Cav-VP1 R°, respectively expressing the structural P1 precursor protein along with the non-structural 3C protein or expressing the structural VP1 protein of the FMDV strain O/FRA/1/2001, were evaluated as novel vaccines against FMD. A strong humoral immune response was elicited in guinea pigs (GP) following immunization with Cav-P1/3C R°, while administration of Cav-VP1 R° did not induce a satisfying antibody response in GP or mice. GP were then used as an experimental model for the determination of the protection afforded by the Cav-P1/3C R° vaccine against challenge with the FMDV strain O Manisa/Turkey/1969. The Cav-P1/3C R° vaccine protected GP from generalized FMD to a similar extent as a high potency double-oil emulsion O Manisa vaccine. The results of the present study show that CAV2-based vector vaccines can express immunogenic FMDV antigens and offer protection against generalized FMD in GP. This suggest that Cav-P1/3C R° FMDV vaccine may protect natural host species from FMD. In combination with an appropriate diagnostic test, the Cav-P1/3C R° FMDV vaccine may also serve as a marker vaccine to differentiate vaccinated from infected animals.
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http://dx.doi.org/10.1016/j.vaccine.2018.02.074DOI Listing
April 2018

Complete Genome Sequences of Five Foot-and-Mouth Disease Viruses of Serotype A Isolated from Cattle in Nigeria between 2013 and 2015.

Genome Announc 2018 Feb 15;6(7). Epub 2018 Feb 15.

Vesicular and Exotic Diseases Unit, Veterinary and Agrochemical Research Centre, Ukkel, Belgium.

The complete genome sequences of 5 foot-and-mouth disease viruses of serotype A are reported here. These viruses originate from outbreaks in northern Nigeria in 2013 to 2015 and belong to the A/AFRICA/G-IV lineage.
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http://dx.doi.org/10.1128/genomeA.00039-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5814479PMC
February 2018