Management of a Focal Introduction of ASF Virus in Wild Boar: The Belgian Experience.

African swine fever (ASF) is a fatal disease of suids that was detected in wild boar in Belgium in September 2018. The measures implemented to stop the spread and eliminate the African swine fever virus consisted of creating restriction zones, organising efficient search and removal of carcasses, constructing wire fences, and depopulating wild boar in the area surrounding the infected zone. The ASF management zone included the infected and the white zones and covered 1106 km² from which 7077 wild boar have been removed. A total of 5338 wild boars have been qPCR-tested and 833 have been detected ASF-positive. The search effort amounted to 60,631 h with a main focus on the infected zone (88%). A total of 277 km of fences have been set up. The main cause of mortality in the infected zone was the virus itself, while hunting, trapping, and night shooting were used together to reduce the wild boar density in the surrounding white zones. After continuous dispersion of the virus until March 2019, the epidemic wave stopped, and the last fresh positive case was discovered in August 2019. Hence, Belgium was declared free of the disease in November 2020.

Epidemiological analyses of African swine fever in the European Union: (September 2020 to August 2021).

This report provides a descriptive analysis of the African swine fever (ASF) Genotype II epidemic in the affected Member States in the EU and two neighbouring countries for the period from 1 September 2020 to 31 August 2021. ASF continued to spread in wild boar in the EU, it entered Germany in September 2020, while Belgium became free from ASF in October 2020. No ASF outbreaks in domestic pigs nor cases in wild boar have been reported in Greece since February 2020. In the Baltic States, overall, there has been a declining trend in proportions of polymerase chain reaction (PCR)-positive samples from wild boar carcasses in the last few years. In the other countries, the proportions of PCR-positive wild boar carcasses remained high, indicating continuing spread of the disease. A systematic literature review revealed that the risk factors most frequently significantly associated with ASF in domestic pigs were pig density, low levels of biosecurity and socio-economic factors. For wild boar, most significant risk factors were related to habitat, socio-economic factors and wild boar management. The effectiveness of different control options in the so-named white zones, areas where wild boar densities have been drastically reduced to avoid further spread of ASF after a new introduction, was assessed with a stochastic model. Important findings were that establishing a white zone is much more challenging when the area of ASF incursion is adjacent to an area where limited control measures are in place. Very stringent wild boar population reduction measures in the white zone are key to success. The white zone needs to be far enough away from the affected core area so that the population can be reduced in time before the disease arrives and the timing of this will depend on the wild boar density and the required population reduction target in the white zone. Finally, establishing a proactive white zone along the demarcation line of an affected area requires higher culling efforts, but has a higher chance of success to stop the spread of the disease than establishing reactive white zones after the disease has already entered in the area.

Schmallenberg virus, cyclical reemergence in the core region: A seroepidemiologic study in wild cervids, Belgium, 2012-2017.

Schmallenberg virus emerged in 2011 in Europe. The epicentre of primordial spreading was the region straddling Germany, the Netherlands and Belgium. One of the key questions is whether the newcomer would establish a lasting presence on the continent. The apparent seroprevalence in southern Belgium wild deer populations was followed for 6 years. Two years of intense circulation were revealed, 2012 and 2016, characterized by a peak seroprevalence in the two studied populations (Capreolus capreolus and Cervus elaphus). Between the peak years and after 2016, apparent seroprevalences declined rapidly among adults and became nil among juveniles. The general pattern of apparent seroprevalence evolution observed is consistent with a cyclic circulation of Schmallenberg virus, similar to what is observed for other Orthobunyaviruses in endemic areas. These data also suggest that wild cervids play no central role in the circulation dynamics of the virus.

Population genetics, invasion pathways and public health risks of the raccoon and its roundworm Baylisascaris procyonis in northwestern Europe.

The geographic range of the zoonotic raccoon roundworm (Baylisascaris procyonis) is expanding together with the range of its host, the raccoon (Procyon lotor). This creates a new public health risk in parts of Europe where this parasite was previously absent. In the Netherlands, a raccoon population is becoming established and incidental findings of B. procyonis have been reported. To assess the risk to public health, the prevalence of B. procyonis was determined in the province of Limburg, where currently the largest Dutch raccoon population is present, as well as in the adjoining region of southern Belgium. Furthermore, genetic methods were employed to assess invasion pathways of both the raccoon and B. procyonis to aid in the development of control measures. Macroscopic analysis of intestinal content and testing of faecal samples were performed to detect B. procyonis adults and eggs. The population genetics of both B. procyonis and its raccoon host were analysed using samples from central and northwestern Europe. B. procyonis was found in 14/23 (61%, 95% CI: 41%-78%) raccoons from Limburg, but was not detected in 50 Belgian raccoons. Genetic analyses showed that the majority of the Dutch raccoons and their roundworms were introduced through ex-captive individuals. As long as free-living raccoon populations originate from captivity, population control methods may be pursued. However, natural dispersal from the border regions will complicate prolonged population control. To reduce the public health risk posed by B. procyonis, public education to increase awareness and adapt behaviour towards raccoons is key.

Epidemiological analysis of African swine fever in the European Union (September 2019 to August 2020).

An update on the African swine fever (ASF) situation in the 10 affected Member States (MS) in the EU and in two neighbouring countries from the 1 September 2019 until the 31 August 2020 is provided. The dynamics of the proportions of PCR- and ELISA-positive samples since the first ASF detection in the country were provided and seasonal patterns were investigated. The impact of the ASF epidemic on the annual numbers of hunted wild boar in each affected MS was investigated. To evaluate differences in the extent of spread of ASF in the wild boar populations, the number of notifications that could be classified as secondary cases to a single source was calculated for each affected MS and compared for the earliest and latest year of the epidemic in the country. To evaluate possible risk factors for the occurrence of ASFV in wild boar or domestic pigs, a literature review was performed. Risk factors for the occurrence of ASF in wild boar in Romanian hunting grounds in 2019 were identified with a generalised linear model. The probability to find at least one PCR-confirmed ASF case in wild boar in a hunting ground in Romania was driven by environmental factors, wild boar abundance and the density of backyard pigs in the hunting ground area, while hunting-related variables were not retained in the final model. Finally, measures implemented in white zones (ASF-free zones that are geographically adjacent to an area where ASF is present in wild boar) to prevent further spread of ASF were analysed with a spatially, explicit stochastic individual-based model. To be effective, the wild boar population in the white zone would need to be drastically reduced before ASF arrives at the zone and it must be wide enough. To achieve the necessary pre-emptive culling targets of wild boar in the white zone, at the start of the establishment, the white zone should be placed sufficiently far from the affected area, considering the speed of the natural spread of the disease. This spread is faster in denser wild boar populations. After a focal ASF introduction, the white zone is always close to the infection hence pre-emptive culling measures in the white zone must be completed in short term, i.e. in a few months.

Evaluation of seven commercial African swine fever virus detection kits and three Taq polymerases on 300 well-characterized field samples.

African swine fever virus (ASFV) is a complex double stranded DNA virus, responsible for a highly infectious and fatal disease in pigs and boars and for important deterioration of animal welfare. Over the last decade, the disease spread to several European and Asian countries causing unprecedented dramatic economic losses in pig industry. In the absence of a vaccine, affected countries rely on trustful diagnostic tests and adapted testing policies to set up control programs to fight against the disease. In this study, we evaluated the sensitivity and specificity of seven commercially available ASFV real-time PCR detection kits and three Taq polymerases on 300 well-characterized wild boar samples collected in Belgium during the 2018-2019 outbreak. This study confirms that all commercial kits and two Taq polymerases are suitable for ASFV detection in diagnostic laboratories. Furthermore, the use of endogenous controls is emphasized when testing field samples harvested on carcasses in an advanced stage of decomposition, in order to avoid false negative results.

Usutu Virus Epizootic in Belgium in 2017 and 2018: Evidence of Virus Endemization and Ongoing Introduction Events.

Wildlife surveillance allowed the monitoring of the zoonotic mosquito-borne Usutu virus (USUV) in birds and bats (Pipistrellus pipistrellus) in southern Belgium in 2017 and 2018. USUV-RNA was detected in 69 birds (of 253) from 15 species, among which 7 species had not previously been reported to be susceptible to the infection. Similarly, 2 bats (of 10) were detected positive by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). USUV-associated lesions were mainly found in Eurasian Blackbirds (Turdus merula), in which USUV antigens were demonstrated by immunohistochemistry in the brain, heart, liver, kidney, intestine, and lung. Partial nonstructural protein 5 gene-based phylogenetic analysis showed several identical or closely related strains from 2016, 2017, and 2018 clustering together within Europe 3 or Africa 3 lineages. Further, one USUV strain detected in a common chaffinch (Fringilla coelebs) manifested a close genetic relationship with the European 1 strains circulating in Hungary and Austria. Our data provide evidence of USUV endemization in southern Belgium in local birds and bats, extension of the host range of the virus and ongoing virus introduction from abroad, likely by migratory birds. Our results highlight the need for vigilance in the forthcoming years toward new virus-associated outbreaks in birds and possible human infections in Belgium.

Multi-host disease management: the why and the how to include wildlife.

In recent years, outbreaks caused by multi-host pathogens (MHP) have posed a serious challenge to public and animal health authorities. The frequent implication of wildlife in such disease systems and a lack of guidelines for mitigating these diseases within wild animal populations partially explain why the outbreaks are particularly challenging. To face these challenges, the French Ministry of Agriculture launched a multi-disciplinary group of experts that set out to discuss the main wildlife specific concepts in the management of MHP disease outbreaks and how to integrate wildlife in the disease management process.This position paper structures the primary specific concepts of wildlife disease management, as identified by the working group. It is designed to lay out these concepts for a wide audience of public and/or animal health officers who are not necessarily familiar with wildlife diseases. The group's discussions generated a possible roadmap for the management of MHP diseases. This roadmap is presented as a cycle for which the main successive step are: step 1-descriptive studies and monitoring; step 2-risk assessment; step 3-management goals; step 4-management actions and step 5-assessment of the management plan. In order to help choose the most adapted management actions for all involved epidemiological units, we integrated a decision-making framework (presented as a spreadsheet). This tool and the corresponding guidelines for disease management are designed to be used by public and health authorities when facing MHP disease outbreaks. These proposals are meant as an initial step towards a harmonized transboundary outbreak response framework that integrates current scientific understanding adapted to practical intervention.

Newly emerged African swine fever virus strain Belgium/Etalle/wb/2018: Complete genomic sequence and comparative analysis with reference p72 genotype II strains.

In a new example of pathogens hopscotching the globe, African swine fever virus hit north-western Europe's wildlife in summer 2018, marking a further spread of a disease that had invaded Central and Eastern Europe recently. The complete genomic sequence of the Belgium/Etalle/wb/2018 virus is reported, with the hope it will provide a valuable tool for tracing geographical spread and biologic evolution of the virus.

First Evidence of Fatal Usutu Virus Natural Infections in an Anatidae, the Common Scoter (Melanitta nigra).

While fatal infections caused by the Usutu virus appeared to concern only passerines (especially the blackbird) and Strigiformes (especially the great gray owl), we report herein that the virus also naturally causes a fatal disease in an anseriforme species, the common scoter (Melanitta nigra).

A novel parvovirus, Roe deer copiparvovirus, identified in Ixodes ricinus ticks.

The family Parvoviridae contains diverse viruses that are capable of infecting a wide range of hosts. In this study, metagenomic sequencing of Ixodes ricinus ticks harvested in 2016 on red deer (Cervus elaphus) and European roe deer (Capreolus capreolus) in Belgium detected a new 6296-bp parvoviral genome. Phylogenetic and sequence analyses showed the new virus belongs to a new species within the Copiparvovirus genus. PCR screening of 4 pools of 10 serum samples from both deer species identified the new copiparvovirus DNA only in roe deer sera. Together, these results are the first evidence of a copiparvovirus in a deer species. Besides its potential pathogenicity to roe deers, the detection of this new virus in ticks raises questions about the possible transmission of parvoviruses by ticks. This report further increases the current knowledge on the evolution and diversity of copiparvoviruses.

Do topography and fruit presence influence occurrence and intensity of crop-raiding by forest elephants (Loxodonta africana cyclotis)?

Crop damage by forest elephants (Loxodonta africana cyclotis) and the resulting human-elephant conflict are issues of great concern for both the conservation of the species and the protection of rural livelihoods in Central Africa. Addressing these problems requires identifying the factors that facilitate or impede crop-raiding by forest elephants. Yet to date, the environmental or anthropogenic factors that influence the occurrence and intensity of crop-raiding by forest elephants are largely unknown. We used a multivariate approach to investigate conditions under which forest elephants raid some fields and not others in the buffer zone of Monts de Cristal National Park (MCNP), Gabon. We first interviewed 121 farmers from 11 villages situated within 10 km of MCNP regarding the occurrence of elephant crop-raiding of their fields. We then collected data on 39 explanatory variables to characterize the agricultural fields. Of these, the most important predictors of elephant raid occurrence of crop damage were presence of fruit trees, elephant deterrents (scarecrows, fire, wire string fences and empty barrels), and field topography. We secondly assessed the effect of stage of crop growth, presence of fruit trees, field topography and presence of elephant deterrents on crop-raiding occurrence and intensity by counting raids and measuring areas of crop damage every week in 17 plantations over 19 weeks in the most elephant-impacted zone of the study area. We found that fruit presence and stage of crop growth led to more intense damage to crops, whereas local deterrents did not inhibit raiding events and crop damage by elephants. We report a tradeoff between non-timber forest products (NTFP) services and crop-raiding by elephants. We show for the first time that steep topography impedes elephant damage to crops with no raids recorded in fields with surrounding slopes greater than 25%. We discuss whether farming on steep fields could be used as a strategy for mitigating crop-raiding to favor human-elephant coexistence and enhance elephant conservation.

Mosquito-borne epornitic flaviviruses: an update and review.

West Nile Virus, Usutu virus, Bagaza virus, Israel turkey encephalitis virus and Tembusu virus currently constitute the five flaviviruses transmitted by mosquito bites with a marked pathogenicity for birds. They have been identified as the causative agents of severe neurological symptoms, drop in egg production and/or mortalities among avian hosts. They have also recently shown an expansion of their geographic distribution and/or a rise in cases of human infection. This paper is the first up-to-date review of the pathology of these flaviviruses in birds, with a special emphasis on the difference in susceptibility among avian species, in order to understand the specificity of the host spectrum of each of these viruses. Furthermore, given the lack of a clear prophylactic approach against these viruses in birds, a meta-analysis of vaccination trials conducted to date on these animals is given to constitute a solid platform from which designing future studies.

Phylogeographic Analysis of African Swine Fever Virus, Western Europe, 2018.

In September 2018, African swine fever in wild boars was detected in Belgium. We used African swine fever-infected spleen samples to perform a phylogenetic analysis of the virus. The causative strain belongs to genotype II, and its closest relatives are viruses previously isolated in Ukraine, Belarus, Estonia, and European Russia.

Moku Virus in Invasive Asian Hornets, Belgium, 2016.

We report the detection of Moku virus in invasive Asian hornets (Vespa velutina nigrithorax) in Belgium. This constitutes an unexpected report of this iflavirus outside Hawaii, USA, where it was recently described in social wasps. Although virulence of Moku virus is unknown, its potential spread raises concern for European honeybee populations.

Widespread activity of multiple lineages of Usutu virus, western Europe, 2016.

In the summer of 2016, Belgium, France, Germany and the Netherlands reported widespread Usutu virus (USUV) activity based on live and dead bird surveillance. The causative USUV strains represented four lineages, of which two putative novel lineages were most likely recently introduced into Germany and spread to other western European countries. The spatial extent of the outbreak area corresponded with R0 values > 1. The occurrence of the outbreak, the largest USUV epizootic registered so far in Europe, allowed us to gain insight in how a recently introduced arbovirus with potential public health implications can spread and become a resident pathogen in a naïve environment. Understanding the ecological and epidemiological factors that drive the emergence or re-emergence of USUV is critical to develop and implement timely surveillance strategies for adequate preventive and control measures. Public health authorities, blood transfusion services and clinicians in countries where USUV was detected should be aware of the risk of possible USUV infection in humans, including in patients with unexplained encephalitis or other neurological impairments, especially during late summer when mosquito densities peak.

Complete Coding Sequences of One H9 and Three H7 Low-Pathogenic Influenza Viruses Circulating in Wild Birds in Belgium, 2009 to 2012.

The complete coding sequences of four avian influenza A viruses (two H7N7, one H7N1, and one H9N2) circulating in wild waterfowl in Belgium from 2009 to 2012 were determined using Illumina sequencing. All viral genome segments represent viruses circulating in the Eurasian wild bird population.

Molecular evidence of Anaplasma phagocytophilum in wild boar (Sus scrofa) in Belgium.

BACKGROUND: Anaplasma phagocytophilum is a tick-borne pathogen of veterinary and human importance. Both ticks as vectors and vertebrates as reservoir hosts are essential for the cycle maintenance of this bacterium. Currently, the whole range of animal species reservoirs for A. phagocytophilum in natural environment is still unknown. Therefore, the aim of this study was to estimate the prevalence of infection with A. phagocytophilum in the wild boar population in southern Belgium. RESULTS: In the frame of a targeted surveillance program, 513 wild boars were sampled during the hunting season 2011. A nested 16S rRNA PCR was used to screen the presence of A. phagocytophilum DNA in spleen of boars. Within 513 samples, 5 (0,97%) were tested PCR positive and identification was confirmed by sequencing. CONCLUSIONS: This study gives the first insight of presence of A. phagocytophilum in wild boars in southern Belgium.