Hepatitis E virus neutralization by porcine serum antibodies.

The consumption of raw or undercooked meat products poses a serious risk for human hepatitis E virus (HEV) infections. In many high-income countries, domestic pigs and wild boars represent the main animal reservoirs for HEV and are usually identified by reverse transcription-PCR and antibody enzyme-linked immunosorbent assay (ELISA). In order to characterize the humoral immune response in more detail, a cell culture-based serum neutralization assay using a culture-adapted HEV strain was established here. Measurement of neutralizing antibodies was only possible after removing the viral quasi-envelope by detergent treatment. Serum samples of 343 wild boars from Northern Germany were first analyzed for anti-HEV IgG using an in-house ELISA, resulting in 19% positive samples. Subsequently, a subset of 41 representative samples was tested with the neutralization assay, and the results correlated well with those obtained by ELISA. Not only the human HEV strain 47832c but also two porcine HEV strains were shown to be neutralized by porcine serum antibodies. Neutralizing activity was also found in samples containing both HEV-specific antibodies and HEV RNA. Testing of serum samples derived from two experimentally infected domestic pigs showed a steep increase in neutralizing activity at 24 or 51 days post infection, dependent on the used infectious dose. The developed assay can be useful for characterization of the humoral immune response after HEV infection and for assessing the efficiency of HEV vaccine candidates.

Heavy Youngsters-Habitat and Climate Factors Lead to a Significant Increase in Body Weight of Wild Boar Females.

As one of the most abundant game species in Europe, European wild boar (Sus scrofa) populations prove highly adaptable to cultivated landscapes. The ongoing process of climate change and the high agricultural yields seem to further optimize the living conditions for this species. In long-term reproduction monitoring, we collected data on the body weight of wild boar females. Over an 18-year period, the body weight of wild boar females increased continuously, then stopped and decreased. It was possible to detect differences between the body weights of animals from forest and agricultural areas. For these areas, differences in body weight development also led to a significant distinction in the onset of puberty. We conclude that, even in a highly cultivated landscape, forested areas provide habitat characteristics that may strongly influence reproduction. Second, with dominant agricultural areas in Germany, wild boar reproduction has been favored in recent decades.

Expanded Diversity and Host Range of Bovine Hepacivirus-Genomic and Serological Evidence in Domestic and Wild Ruminant Species.

The hepatitis C virus (HCV)-related bovine hepacivirus (BovHepV) can cause acute as well as persistent infections in cattle. The true clinical relevance of the virus is not yet known. As reliable antibody detection methods are lacking and prevalence studies have only been conducted in cattle and few countries to date, the true distribution, genetic diversity, and host range is probably greatly underestimated. In this study, we applied several RT-PCR methods and a nano-luciferase-based immunoprecipitation system (LIPS) assay to analyze bovine serum samples from Bulgaria as well as wild ruminant sera from Germany and the Czech Republic. Using these methods, BovHepV infections were confirmed in Bulgarian cattle, with viral genomes detected in 6.9% and serological reactions against the BovHepV NS3 helicase domain in 10% of bovine serum samples. Genetic analysis demonstrated co-circulation of highly diverse BovHepV strains in Bulgarian cattle, and three novel BovHepV subtypes within the genotype 1 could be defined. Furthermore, application of a nested RT-PCR led to the first description of a BovHepV variant (genotype 2) in a wild ruminant species. The results of this study significantly enhance our knowledge of BovHepV distribution, genetic diversity, and host range.

Isolation of 15 hepatitis E virus strains lacking ORF1 rearrangements from wild boar and pig organ samples and efficient replication in cell culture.

As a zoonotic pathogen, the hepatitis E virus (HEV) leads to numerous infections in humans with different clinical manifestations. Especially genotype 3, as causative agent of a foodborne zoonosis, is transmitted to humans by ingestion of undercooked or raw meat containing liver from HEV-infected animals. Although the virus' prevalence and dissemination in hosts like wild boar and pig have been well characterized, HEV is greatly understudied on a molecular level and reliable cell culture models are lacking. For this reason, the present study concentrated on the isolation and subsequent characterization of porcine HEV from tissue samples derived from wild boar and domestic pigs: 222 wild boars hunted in Northern Germany were investigated for the presence of HEV RNA with a detection rate of 5.9%. Three additional HEV-positive wild boar liver samples as well as an HEV-positive spleen and a positive kidney from domestic pigs were included. After inoculation of positive samples onto the human hepatoma cell line PLC/PRF/5, cells were grown for several weeks. Successful isolation was confirmed by RT-qPCR, virus passage, immunofluorescence staining and titration. Overall, 15 strains from a total of 18 RNA-positive organ samples could be obtained and viral loads >109  RNA copies/ml were measured in cell culture supernatants. Accordingly, 83.3% of the HEV RNA-positive samples contained infectious hepatitis E viral particles and therefore must be considered as a potential source for human infections. Phylogenetic analyses revealed that all isolated strains belong to genotype 3. Further genetic characterization showed a high degree of sequence variability, but no sequence insertions, in the hypervariable region within the open reading frame 1.

Zoning has little impact on the seasonal diel activity and distribution patterns of wild boar (Sus scrofa) in an UNESCO Biosphere Reserve.

Understanding the spatio-temporal distribution of ungulates is important for effective wildlife management, particularly for economically and ecologically important species such as wild boar (Sus scrofa). Wild boars are generally considered to exhibit substantial behavioral flexibility, but it is unclear how their behavior varies across different conservation management regimes and levels of human pressure. To analyze if and how wild boars adjust their space use or their temporal niche, we surveyed wild boars across the core and buffer zones (collectively referred to as the conservation zone) and the transition zone of a biosphere reserve. These zones represent low and high levels of human pressure, respectively. Specifically, we employed a network of 53 camera traps distributed in the Schaalsee UNESCO Biosphere Reserve over a 14-month period (19,062 trap nights) and estimated circadian activity patterns, diel activity levels, and occupancy of wild boars in both zones. To account for differences in environmental conditions and day length, we estimated these parameters separately for seven 2-month periods. Our results showed that the wild boars were primarily nocturnal, with diurnal activity occurring dominantly during the summer months. The diel activity patterns in the two zones were very similar overall, although the wild boars were slightly less active in the transition zone than in the conservation zone. Diel activity levels also varied seasonally, ranging from 7.5 to 11.0 h day-1, and scaled positively with the length of the night (R 2 = 0.66-0.67). Seasonal occupancy estimates were exceptionally high (point estimates ranged from 0.65 to 0.99) and similar across zones, suggesting that the wild boars used most of the biosphere reserve. Overall, this result suggests that different conservation management regimes (in this case, the zoning of a biosphere reserve) have little impact on wild boar behavior. This finding is relevant for wildlife management in protected areas where possibly high wild boar densities could interfere with conservation goals within these areas and those of agricultural land use in their vicinity.

How Do Hunters Hunt Wild Boar? Survey on Wild Boar Hunting Methods in the Federal State of Lower Saxony.

High wild boar population densities lead to demands for a population reduction to avoid crop damages or epidemic diseases. Along with biological studies, a better understanding of the human influence on wildlife and on wildlife management is important. We conducted inquiries on hunting methods and on hunters' attitudes in the Federal State of Lower Saxony, Germany, to better understand hunting strategies and the influence on increasing wild boar population, as well as to underpin game management concepts. Single hunt, especially at bait, is still the most widely used method for hunting wild boar. The proportion of drive hunts within the hunting bag is increasing. The proportions of hunting methods vary regionally due to wild boar densities, geographical features (vegetation, terrain, etc.) and hunters' practices. Hunters increased the proportion of conjoint hunts on wild boar. Baiting remains an important hunting method in wild boar management and the proportion of drive hunts should be fostered. Private hunting is important for wild boar management, although it is just insufficient. Additionally, administrative wildlife managers are recommended for the near future as coordinators of wild boar management, and as such, could manage hunting, the incorporation of regional conditions and investigating hunters' attitudes and abilities.

An approach to assess stress in response to drive hunts using cortisol levels of wild boar (Sus scrofa).

Hunting can easily be linked to stress in wildlife. Drive hunts performed two to three times in one area during the respective hunting period, are thought to decrease the pressure hunting places on wildlife. Nevertheless, the expression of cortisol-one of the main mammalian stress hormones-is considered to have negative impacts on animals' well-being if expressed excessively, which may occur during some (especially repeated) hunting events. We explored the effect of drive hunts on cortisol levels in wild boar in Lower Saxony, Germany, compared these cortisol levels to reference values given by a similar study, and investigated the effect of age, sex, and pregnancy. Blood collected from wild boar shot on drive hunts was analysed using a radioimmunoassay. As expected, we observed elevated cortisol levels in all samples, however, we still found significant differences between age groups and sexes, as well as an influence of pregnancy on cortisol levels. The effect of drive hunts on cortisol levels appears to be weaker than predicted, while the effects of other variables, such as sex, are distinct. Only half of the evaluated samples showed explicitly increased cortisol levels and no significant differences were found between sampling months and locations. Group living animals and pregnant females showed significantly higher cortisol levels. The impact of hunting is measurable but is masked by natural effects such as pregnancy. Thus, we need more information on stress levels in game species.

Differences in wild boar spatial behaviour among land uses and management scenarios in Mediterranean ecosystems.

The ubiquitous wild boar is causing diverse and growing conflicts of socio-ecological and economic relevance worldwide. For that reason, knowledge of its spatial ecology is crucial to designing effective management programmes. But this knowledge is scarce in Mediterranean areas with mixed land uses. We describe the spatial ecology and habitat selection of 41 adult wild boar monitored using GPS collars and analyse the effects of sex and the period (food shortage period, hunting season and food abundance period) under different land uses (protected areas, mixed farms and fenced hunting estates). The spatial ecology of wild boar was characterised by marked temporality, mediated by sex and the land uses in the area. The activity (ACT), daily range (DR) and home range (HR) were higher for males than females, and in mixed farms versus fenced hunting estates, while the lowest values were obtained in protected areas. These effects were more marked for ACT and DR (movement) than HR. The selection of scrublands and avoidance of woodlands was observed where drive hunt events occur (mixed farms and fenced estates), but not in the protected areas. The differences in the requirements, reproductive behaviour and, interestingly, response to disturbance according to sex may explain this dissimilar behaviour. Disturbance originated higher movement rates and the selection of sheltering land cover as a refuge during the hunting season. This information is useful for designing species monitoring and management programmes; including both preventive and reactive actions in response to events such as outbreaks of African swine fever and agricultural damage produced by wild boar in Europe.

Precocious puberty in male wild boars: a possible explanation for the dramatic population increase in Germany and Europe.

BACKGROUND: The wild boar population in Europe is steadily growing, one of the reasons for this increase probably being the high reproductive potential of this large mammal. Population management is important to stabilise wild boar numbers and a great deal of attention is focusing on the reasons, which might contribute to the high reproductive rates. Understanding the timing of puberty attainment provides information required for proper management practices. Knowledge of the earliest expected time of sexual maturation in male wild boars is limited, research being mostly focused on females. Previous hunting references indicate that sexual maturity in males occurs in the second year after birth. In contrast, male domestic pigs become sexually mature from about seven months of age. Thus, aims of this study were to investigate (1) whether there is a physiological ability for reproduction also in male wild boars of a younger age and (2) whether the body weight of wild boar males has a more important role than age in driving the maturation of the testis. METHODS: Male wild boar individuals were sampled during hunting drives in the eastern part of Lower Saxony in Germany. Testes with epididymides from 74 males were collected and prepared for histological examination and immunohistochemistry. The reproductive status could be ascertained based on development/occurrence of different germ cell populations using histology and based on the immunohistochemical detection of the anti-Müllerian hormone and androgen receptor. RESULTS: In this study, male wild boars aged nine to ten months already passed puberty and were able to reproduce if they had reached the appropriate body condition of about 29 kg dressed weight. Immunopositivity to the anti-Müllerian hormone in Sertoli cells was evident only in prepubertal animals and decreased with the onset of puberty. No immunoreaction was evident at postpuberty. The androgen receptor was detected in Sertoli cells, peritubular cells and Leydig cells, surprisingly already in Sertoli cells of prepubertal wild boars as well depending on body weight. Moreover, two-thirds of young males aged about ten months were precociously reproductively mature, showing histologically the presence of spermatozoa in testes and epididymides. CONCLUSIONS: As piglets are mostly born in spring, also these young male individuals could target the heat of female wild boars in the winter months, resulting in the observed population increase. Therefore, a reduction in wild boar numbers should also focus on piglets of both sexes.

Behavioural patterns of free roaming wild boar in a spatiotemporal context.

Although the almost worldwide distributed wild boar Sus scrofa is a well-studied species, little is known about the behaviour of autochthonous, free living wild boar in a spatiotemporal context which can help to better understand wild boar in conflict terms with humans and to find solutions. The use of camera traps is a favourable and non-invasive method to study them. To observe natural behaviour, 60 camera traps were placed for three months in a state forest of 17.8 km2 in the region of the Luneburg Heath in northern Germany. In this area wild boar, roe deer, red deer, wolves and humans are common. The cameras recorded 20 s length video clips when animals passed the detection zone and could be triggered again immediately afterwards. In total 38 distinct behavioural elements were observed, which were assigned to one of seven behavioural categories. The occurrence of the behavioural categories per day was evaluated to compare their frequencies and see which are more essential than others. Generalised Additive Models were used to analyse the occurrence of each behaviour in relation to habitat and activity time. The results show that essential behavioural categories like foraging behaviour, locomotion and vigilance behaviour occurred more frequently than behaviour that "just" served for the well-being of wild boar. These three behavioural categories could be observed together mostly in the night in broad-leaved forests with a herb layer of 50-100%, comfort behaviour occurred mostly at the ponds in coniferous forest. It is also observable that the behavioural categories foraging and comfort behaviour alternated several times during the night which offers the hypothesis that foraging is mostly followed by comfort behaviour. These findings pave the way towards implementing effective control strategies in the wild and animal welfare in captivity.

Human-wildlife coexistence in a changing world.

Human-wildlife conflict (HWC) is a key topic in conservation and agricultural research. Decision makers need evidence-based information to design sustainable management plans and policy instruments. However, providing objective decision support can be challenging because realities and perceptions of human-wildlife interactions vary widely between and within rural, urban, and peri-urban areas. Land users who incur costs through wildlife argue that wildlife-related losses should be compensated and that prevention should be subsidized. Supporters of human-wildlife coexistence policies, such as urban-dwelling people, may not face threats to their livelihoods from wildlife. Such spatial heterogeneity in the cost and benefits of living with wildlife is germane in most contemporary societies. This Special Section features contributions on wildlife-induced damages that range from human perspectives (land use, psychology, governance, local attitudes and perceptions, costs and benefits, and HWC and coexistence theory) to ecological perspectives (animal behavior). Building on current literature and articles in this section, we developed a conceptual model to help frame HWC and coexistence dimensions. The framework can be used to determine damage prevention implementation levels and approaches to HWC resolution. Our synthesis revealed that inter- and transdisciplinary approaches and multilevel governance approaches can help stakeholders and institutions implement sustainable management strategies that promote human-wildlife coexistence.

Coexistencia Humano - Vida Silvstre en un Mundo Cambiante Resumen El conflicto humano - vida silvestre (HWC) es un tema muy importante para la investigación agrícola y de la conservación. Los tomadores de decisiones necesitan información basada en evidencias para diseñar planes de manejo sustentable e instrumentos políticos. Sin embargo, proporcionar un apoyo objetivo para las decisiones puede ser un reto ya que las realidades y percepciones de las interacciones humano - vida silvestre varían enormemente entre y dentro de las áreas rurales, urbanas y peri-urbanas. Los usuarios de terrenos que incurren en costos debido a la vida silvestre argumentan que las pérdidas relacionadas a la vida silvestre deberían ser compensadas y que la prevención debería estar subsidiada. Es probable que quienes apoyan las políticas de coexistencia entre humanos y vida silvestre, como los habitantes de zonas urbanas, no enfrenten una amenaza a su medio de subsistencia debido a la vida silvestre. Dicha heterogeneidad espacial en el costo y beneficio de cohabitar con la vida silvestre es relevante en la mayoría de las sociedades contemporáneas. Esta Sección Especial presenta contribuciones sobre daños inducidos por vida silvestre que van desde las perspectivas humanas (uso de suelo, psicología, gobierno, actitudes y percepciones locales, costo y beneficio y la teoría del conflicto y la coexistencia humano-vida silvestre) hasta las perspectivas ecológicas (comportamiento animal). A partir de los artículos y la literatura actuales en esta sección desarrollamos un modelo conceptual para ayudar a estructurar los alcances del HWC y de la coexistencia. El marco de trabajo puede usarse para determinar los niveles y estrategias de implementación de la prevención del daño a la solución del conflicto humano - fauna. Nuestra síntesis reveló que las estrategias inter- y transdisciplinarias y las estrategias de gobierno multiniveles pueden ayudar a que los actores y las instituciones implementen estrategias de manejo sustentable para promover la coexistencia entre los humanos y la vida silvestre.

Genetic variability of porcine pegivirus in pigs from Europe and China and insights into tissue tropism.

Pegiviruses belong to the family Flaviviridae and have been found in humans and other mammalian species. To date eleven different pegivirus species (Pegivirus A-K) have been described. However, little is known about the tissue tropism and replication of pegiviruses. In 2016, a so far unknown porcine pegivirus (PPgV, Pegivirus K) was described and persistent infection in the host, similar to human pegivirus, was reported. In this study, qRT-PCR, phylogenetic analyses and fluorescence in situ hybridization (FISH) were implemented to detect and quantify PPgV genome content in serum samples from domestic pigs from Europe and Asia, in tissue and peripheral blood mononuclear cell (PBMC) samples and wild boar serum samples from Germany. PPgV was detectable in 2.7% of investigated domestic pigs from Europe and China (viral genome load 2.4 × 102 to 2.0 × 106 PPgV copies/ml), while all wild boar samples were tested negative. Phylogenetic analyses revealed pairwise nucleotide identities >90% among PPgVs. Finally, PPgV was detected in liver, thymus and PBMCs by qRT-PCR and FISH, suggesting liver- and lymphotropism. Taken together, this study provides first insights into the tissue tropism of PPgV and shows its distribution and genetic variability in Europe and China.

Regulating wild boar populations is "somebody else's problem"! - Human dimension in wild boar management.

As a part of the ongoing game survey of the German federal state of Lower Saxony (WTE), we conducted inquiries into wild boar management and distribution, as well as hunters' attitudes, in order to determine the reasons for the increase of wild boar populations and to inform our game management strategy. According to hunters' reports within the WTE, increases in distribution and population continue and a reduction of the wild boar population has been deemed necessary on a large scale. In the home region, however, it seems to be "somebody else's problem" (SEP), according to hunters' opinions. The majority of hunters are not able to regulate the population and this could be a reason that wild boar numbers continue to increase. Cooperation and comprehensive hunting with efficient hunting methods seems to be the most promising solution, as non-hunting methods are unpopular amongst hunters. The hunters seem to be aware of the problems, solutions and contributing factors; however, most hunters do not feel responsible and see the management of wild boar, again, as a SEP. Regional conditions, as well as hunters' willingness and capacity to manage wild boar will have to be incorporated into management concepts.

The influence of environmental and physiological factors on the litter size of wild boar (Sus scrofa) in an agriculture dominated area in Germany.

The wild boar population has increased enormously in all of Europe over the last decades and caused problems like crop damage, transmission of diseases, and vehicle accidents. Therefore, it is necessary to investigate the underlying causes of this increase in order to be able to manage populations effectively. The purpose of this study was to analyse how environmental (food and climate) and physiological factors (maternal weight and age) as well as hunting and population density influence the litter size of wild boar populations in Northern Germany. The mean litter size in the studied population for the whole period was 6.6 (range 1­12), which is one of the highest in all of Europe. Litter size was positively influenced by maternal body weight, higher mast yield of oak as well as higher temperature in combination with higher precipitation in summer. Only higher temperature or only higher precipitation in summer however had a negative effect on litter size production. Probably,weather and food conditions act via maternal bodyweight on the litter size variation in wild boar. Hunting as well a s population density did not affect the litter size variation in this study which might indicate that wild boar population did not reach carrying capacity yet.