Niche overlap across landscape variability in summer between two large herbivores using eDNA metabarcoding.

Understanding the relationship between a species feeding strategy and its environment (trophic ecology) is critical to assess environmental requirements and improve management policies. However, measuring trophic interactions remains challenging. Among the available methods, quantifying the plant composition of a species' diet indicates how species use their environment and their associated niche overlap. Nevertheless, most studies focusing on herbivore trophic ecology ignore the influence that landscape variability may have. Here, we explored how landscape variability influences diet composition through niche overlap. We used eDNA metabarcoding to quantify the diet composition of two large herbivores of the Bialowieza Forest, red deer (Cervus elaphus) and European bison (Bison bonasus) to investigate how increasing habitat quality (i.e. higher abundance of deciduous forage species) and predation risk (i.e. density of wolf in the area) influence their diet composition and niche partitioning. Our findings indicate diet composition is non-homogeneous across the landscape, both within and between species. Red deer showed greater diet variability and lower niche overlap within species compared to bison. We detected a reduction of niche overlap for red deer with increasing predation risk, leading to more dissimilar diets, suggesting their feeding behaviour is affected by wolf presence. This correlation was not found for bison, which are rarely predated by wolf. Higher habitat quality was associated with higher niche overlap only within bison, probably due to their suboptimal feeding strategy as browsers. These results show the importance of integrating environment-induced diet variation in studies aimed at determining the landscape usage or niche overlap of a species.

Badgers remain fearless in the face of simulated wolf presence near their setts.

Many mesocarnivores are fossorial and use burrow systems to avoid predators. But fossorial animals cannot stay safely underground forever; they must also risk emerging overground to forage and find mates. To make this trade-off effectively and maximise their own fitness, it is imperative they assess how risk varies in space and time and adapt their denning behaviour accordingly. We used the badger in Bialowieza Forest, Poland, as a model for investigating how the denning behaviour of a fossorial mesocarnivore varies in response to short-term large carnivore risk. To this end, we experimentally simulated perceived wolf presence outside 10 badger setts using audio playbacks of wolves (their howls). We assayed two behavioural measures of fear: badger emergence time from setts on the day playbacks were broadcast and their presence in setts on the day after. We found that neither badger emergence time nor next-day sett use varied in response to wolf playbacks. The results of the present study contrast with a previous study of ours that found badgers used setts in areas with high landscape level perceived wolf risk less often than those in lower-risk areas. Together, these papers' findings suggest that different spatiotemporal scales of perceived risk can have differential effects on badger behaviour. We conclude that rather than take risk avoidance measures at all risky times and places, badgers likely display a diversity of reactions to large carnivore presence that depend on the context and spatiotemporal scale of the risk being perceived.

Wolf risk fails to inspire fear in two mesocarnivores suggesting facilitation prevails.

Large carnivores not only supress mesocarnivores via killing and instilling fear, but also facilitate them through carrion provisioning. Hence, mesocarnivores frequently face a trade-off between risk avoidance and food acquisition. Here we used the raccoon dog and red fox in Bialowieza Forest, Poland as models for investigating how large carnivores shape mesocarnivore foraging behaviour in an area with widespread large carnivore carrion provisioning. Using a giving up density experiment we quantified mesocarnivore foraging responses to wolf body odour across a landscape-scale gradient in wolf encounter rates. At locations with higher wolf encounter rates, raccoon dogs depleted feeding trays more than at plots with lower wolf encounter rates. Simulating wolf presence by adding wolf body odour caused raccoon dogs to deplete feeding trays more at locations with low wolf encounter rates, but less at locations with high wolf encounter rates. Fox foraging costs did not vary with the application of wolf body odour or wolf encounter rates. The frequency that the mesocarnivores visited experimental foraging patches was unaffected by wolf body odour or landscape level encounter rates. These results provide further evidence that large carnivore suppression can play a subordinate role to facilitation in determining mesocarnivore behaviour. The varying raccoon dog response to wolf odour across the landscape-scale gradient in wolf encounter rates shows how mesocarnivore-large carnivore interactions can be context-dependent. We suggest that rather than testing the effects of single risk cues on prey behaviour, future studies should focus on understanding how context modifies the ecological impacts of large carnivores.

Emerging infectious disease triggered a trophic cascade and enhanced recruitment of a masting tree.

There are several mechanisms that allow plants to temporarily escape from top-down control. One of them is trophic cascades triggered by top predators or pathogens. Another is satiation of consumers by mast seeding. These two mechanisms have traditionally been studied in separation. However, their combined action may have a greater effect on plant release than either process alone. In 2015, an outbreak of a disease (African swine fever, ASF) caused a crash in wild boar (Sus scrofa) abundance in Bialowieza Primeval Forest. Wild boar are important consumers of acorns and are difficult to satiate relative to less mobile granivores. We hypothesized that the joint action of the ASF outbreak and masting would enhance regeneration of oaks (Quercus robur). Data from ungulate exclosures demonstrated that ASF led to reduction in acorn predation. Tree seedling data indicated that oak recruitment increased twofold relative to pre-epidemic period. Our results showed that perturbations caused by wildlife disease travel through food webs and influence forest dynamics. The outbreak of ASF acted synergistically with masting and removed herbivore top-down control of oaks by mobile consumers. This illustrates that the ASF epidemic that currently occurs across Europe can have broad effects on forest dynamics.

Olfactory cues of large carnivores modify red deer behavior and browsing intensity.

This study examined the effect of perceived predation risk imposed by lynx (Lynx lynx) and wolf (Canis lupus) on red deer (Cervus elaphus) foraging behavior under experimental conditions. We hypothesized that in response to large carnivore scent red deer would increase their vigilance, although reducing the frequency and duration of visits to foraging sites. Consequently, browsing intensity on tree saplings was expected to decrease, whereas a higher proportion of more preferred species was expected to be browsed to compensate for higher foraging costs. We expected stronger responses towards the ambush predator lynx, compared with the cursorial predator wolf. These hypotheses were tested in a cafeteria experiment conducted within three red deer enclosures, each containing four experimental plots with olfactory cues of wolf, lynx, cow, and water as control. On each plot, a camera trap was placed and browsing intensity was measured for one consecutive week, repeated three times. Red deer reduced their visitation duration and browsing intensity on plots with large carnivore scent. Despite red deer showing a clear preference for certain tree species, the presence of large carnivore scent did not change selectivity towards different tree species. Contrary to our hypothesis, we found more pronounced effects of wolf (cursorial) compared with lynx (ambush). This study is the first to experimentally assess the perceived risk effects on the red deer foraging behavior of large carnivores differing in hunting modes. Our findings provide insights into the role of olfactory cues in predator-prey interactions and how they can modify fine-scale herbivore-plant interactions.

Disease-Induced Mortality Outweighs Hunting in Causing Wild Boar Population Crash After African Swine Fever Outbreak.

African swine fever (ASF) has been spreading in the Eurasian continent for more than 10 years now. Although the course of ASF in domestic pigs and its negative economic impact on the pork industry are well-known, we still lack a quantitative assessment of the impact of ASF on wild boar (Sus scrofa) populations under natural conditions. Wild boar is not only a reservoir for ASF; it is also one of the key wildlife species affecting structure and functioning of ecosystems. Therefore, knowledge on how ASF affects wild boar populations is crucial to better predict ecosystem response and for the design of scientific-based wild boar management to control ASF. We used a long-term camera trap survey (2012-2017) from the Bialowieza Primeval Forest (BPF, Poland), where an ASF outbreak occurred in 2015, to investigate the impact of the disease on wild boar population dynamics under two contrasting management regimes (hunted vs. non-hunted). In the hunted part of BPF ("managed area"), hunting was drastically increased prior and after the first ASF case occurred (March 2015), whereas inside the National Park, hunting was not permitted ("unmanaged area," first detected case in June 2015). Using a random encounter model (REM), we showed that the density and abundance of wild boar dropped by 84 and 95% within 1 year following ASF outbreak in the unmanaged and managed area, respectively. In the managed area, we showed that 11-22% additional mortality could be attributed to hunting. Our study suggests that ASF-induced mortality, by far, outweighs hunting-induced mortality in causing wild boar population decline and shows that intensified hunting in newly ASF-infected areas does not achieve much greater reduction of population size than what is already caused by the ASF virus.

Behavioral response of naïve and non-naïve deer to wolf urine.

Large carnivores are recolonizing many regions in Europe, where their ungulate prey have lived without them for >150 years. Whether the returning large carnivores will modify ungulate behavior and indirectly affect lower trophic levels, depends on the ability of ungulates to recognize risk based on past encounters and cues indicating carnivore presence. In two case studies, we tested, by means of camera trapping, the behavioral response of deer to wolf urine. The first case study was in the Netherlands where deer (still) live in absence of wolves, and the second in Poland with long-term wolf presence. As controls we used water (no scent) and all-purpose soap (unfamiliar scent). Deer vigilance level on control plots was 20% in both case studies indicating that wolf occupancy per se does not lead to a consistent difference in behavior. Placing wolf urine did not significantly affect deer behavior in either the wolf-absent or the wolf-present area. More intense cues, or a combination of cues, are likely needed to affect deer behavior. Moreover, we found an unexpected reaction of deer towards all-purpose soap of reduced foraging (and tendency for increased vigilance) in the wolf-present area, whereas it did not affect deer behavior in the wolf-absent area. We hypothesize that deer associate all-purpose soap with human presence, causing no response in human-dominated landscapes (the Netherlands), but triggering a behavioral reaction in more remote areas (Poland). This illustrates attention should be paid to controls used in scent experiments as they may be associated differently than intended.

Linking spatial patterns of terrestrial herbivore community structure to trophic interactions.

Large herbivores influence ecosystem functioning via their effects on vegetation at different spatial scales. It is often overlooked that the spatial distribution of large herbivores results from their responses to interacting top-down and bottom-up ecological gradients that create landscape-scale variation in the structure of the entire community. We studied the complexity of these cascading interactions using high-resolution camera trapping and remote sensing data in the best-preserved European lowland forest, Bialowieza Forest, Poland. We showed that the variation in spatial distribution of an entire community of large herbivores is explained by species-specific responses to both environmental bottom-up and biotic top-down factors in combination with human-induced (cascading) effects. We decomposed the spatial variation in herbivore community structure and identified functionally distinct landscape-scale herbivory regimes ('herbiscapes'), which are predicted to occur in a variety of ecosystems and could be an important mechanism creating spatial variation in herbivory maintaining vegetation heterogeneity.

Implications of shared predation for space use in two sympatric leporids.

Spatial variation in habitat riskiness has a major influence on the predator-prey space race. However, the outcome of this race can be modulated if prey shares enemies with fellow prey (i.e., another prey species). Sharing of natural enemies may result in apparent competition, and its implications for prey space use remain poorly studied. Our objective was to test how prey species spend time among habitats that differ in riskiness, and how shared predation modulates the space use by prey species. We studied a one-predator, two-prey system in a coastal dune landscape in the Netherlands with the European hare (Lepus europaeus) and European rabbit (Oryctolagus cuniculus) as sympatric prey species and red fox (Vulpes vulpes) as their main predator. The fine-scale space use by each species was quantified using camera traps. We quantified residence time as an index of space use. Hares and rabbits spent time differently among habitats that differ in riskiness. Space use by predators and habitat riskiness affected space use by hares more strongly than space use by rabbits. Residence time of hare was shorter in habitats in which the predator was efficient in searching or capturing prey species. However, hares spent more time in edge habitat when foxes were present, even though foxes are considered ambush predators. Shared predation affected the predator-prey space race for hares positively, and more strongly than the predator-prey space race for rabbits, which were not affected. Shared predation reversed the predator-prey space race between foxes and hares, whereas shared predation possibly also released a negative association and promoted a positive association between our two sympatric prey species. Habitat riskiness, species presence, and prey species' escape mode and foraging mode (i.e., central-place vs. noncentral-place forager) affected the prey space race under shared predation.

Lessons from Bialowieza Forest on the history of protection and the world's first reintroduction of a large carnivore.

Understanding how the relationships between large carnivores and humans have evolved and have been managed through centuries can provide relevant insights for wildlife conservation. The management history of many large carnivores has followed a similar pattern, from game reserved for nobility, to persecuted pests, to conservation targets. We reconstructed the history of brown bear (Ursus arctos) management in Bialowieza Forest (Poland and Belarus) based on a detailed survey of historical literature and Russian archives. From the end of the Middle Ages to the end of 18th century, the brown bear was considered "animalia superiora" (i.e., game exclusively reserved for nobility and protected by law). Bears, also a source of public entertainment, were not regarded as a threat. Effective measures to prevent damages to traditional forest beekeeping were already in practice. In the beginning of 19th century, new game-management approaches allowed most forest officials to hunt bears, which became the primary target of hunters due to their valuable pelt. This, together with an effective anticarnivore policy enhanced by bounties, led to bear extirpation in 1879. Different approaches to scientific game management appeared (planned extermination of predators and hunting levels that would maintain stable populations), as did the first initiatives to protect bears from cruel treatment in captivity. Bear reintroduction in Bialowieza Forest began in 1937 and represented the world's first reintroduction of a large carnivore motivated by conservation goals. The outbreak of World War II spoiled what might have been a successful project; reproduction in the wild was documented for 8 years and bear presence for 13. Soft release of cubs born in captivity inside the forest but freely roaming with minimal human contact proved successful. Release of captive human-habituated bears, feeding of these bears, and a lack of involvement of local communities were weaknesses of the project. Large carnivores are key components of ecosystem-function restoration, and site-specific histories provide important lessons in how to preserve them for the future.

Mesopredator behavioral response to olfactory signals of an apex predator.

Olfactory signals constitute an important mechanism in interspecific interactions, but little is known regarding their role in communication between predator species. We analyzed the behavioral responses of a mesopredator, the red fox (Vulpes vulpes), to an olfactory cue (scat) of an apex predator, the lynx (Lynx lynx) in Bialowieza Primeval Forest, Poland, using video camera traps. Red fox visited sites with scats more often than expected and the duration of their visits was longer at scat sites than at control sites (no scat added). Vigilant behavior, sniffing and scent marking (including over-marking) occurred more often at scat sites compared to control sites, where foxes mainly passed by. Vigilance was most pronounced during the first days of the recordings. Red fox behavior was also influenced by foxes previously visiting scat sites. They sniffed and scent marked (multiple over-marking) more frequently when the lynx scat had been over-marked previously by red fox. Fox visits to lynx scats may be seen as a trade-off between obtaining information on a potential food source (prey killed by lynx) and the potential risk of predation by an apex predator.

Brown world forests: increased ungulate browsing keeps temperate trees in recruitment bottlenecks in resource hotspots.

Plant biomass consumers (mammalian herbivory and fire) are increasingly seen as major drivers of ecosystem structure and function but the prevailing paradigm in temperate forest ecology is still that their dynamics are mainly bottom-up resource-controlled. Using conceptual advances from savanna ecology, particularly the demographic bottleneck model, we present a novel view on temperate forest dynamics that integrates consumer and resource control. We used a fully factorial experiment, with varying levels of ungulate herbivory and resource (light) availability, to investigate how these factors shape recruitment of five temperate tree species. We ran simulations to project how inter- and intraspecific differences in height increment under the different experimental scenarios influence long-term recruitment of tree species. Strong herbivore-driven demographic bottlenecks occurred in our temperate forest system, and bottlenecks were as strong under resource-rich as under resource-poor conditions. Increased browsing by herbivores in resource-rich patches strongly counteracted the increased escape strength of saplings in these patches. This finding is a crucial extension of the demographic bottleneck model which assumes that increased resource availability allows plants to more easily escape consumer-driven bottlenecks. Our study demonstrates that a more dynamic understanding of consumer-resource interactions is necessary, where consumers and plants both respond to resource availability.

Transition from traditional to modern forest management shaped the spatial extent of cattle pasturing in Bialowieza Primeval Forest in the nineteenth and twentieth centuries.

Pasturing of livestock in forests has had profound consequences for Europe's landscapes. In Bialowieza Primeval Forest (BPF), cattle pasturing was a part of traditional forest use that ceased only in the second half of the twentieth century. We collected information on the institutional changes governing forest cattle pasturing and the changes in spatial extent of cattle presence in BPF in last two centuries and information on cattle numbers and their impact on forest regeneration. The spatial extent of cattle pasturing was highly variable, with the distribution of grazing areas frequently changing. Forest near villages (constituting less than 10 % of the area) was most often used for cattle grazing during continued longer time periods. Historical data showed that cattle have had a clear impact on forest regeneration. However, the frequent changes that occurred in the extent of cattle grazing indicate that their impact occurred locally, was smaller in other less intensively used areas, and in the forest as a whole.

Cyclical succession in grazed ecosystems: the importance of interactions between different-sized herbivores and different-sized predators.

Body size of vertebrate herbivores is strongly linked to other life history traits, most notably (1) tolerance of low quality forage and (2) vulnerability to predation, which both impact the composition and dynamics of natural communities. However, no study has thus far explored how the combination of these two body-size related traits affects the long-term composition and dynamics of the herbivore and plant communities. We made a simple model of ordinary differential equations and simulated a grassland system with three herbivore species (small, medium, large) and two predator species (small, large) to investigate how the combination of low-quality tolerance and predation-vulnerability structure the herbivore and plant community. We found that facilitation and competition between different-sized herbivores and predation by especially small predators stimulate coexistence among herbivore species. Furthermore, the interaction between different-sized herbivores and predators generated cyclical succession in the plant community, i.e. alternating periods of short vegetation dominated by high-quality plants, with periods of tall vegetation dominated by low-quality plants. Our results suggest that cyclical succession in plant communities is more likely to occur when a predator predominantly preys on small herbivore species. Large predators also play an important role, as their addition relaxed the set of conditions under which cyclical succession occurred. Consequently, our model predictions suggest that a diverse predator community plays an important role in the long-term dynamics and maintenance of diversity in both the herbivore and plant community.

What cues do ungulates use to assess predation risk in dense temperate forests?

Anti-predator responses by ungulates can be based on habitat features or on the near-imminent threat of predators. In dense forest, cues that ungulates use to assess predation risk likely differ from half-open landscapes, as scent relative to sight is predicted to be more important. We studied, in the Bialowieza Primeval Forest (Poland), whether perceived predation risk in red deer (Cervus elaphus) and wild boar (Sus scrofa) is related to habitat visibility or olfactory cues of a predator. We used camera traps in two different set-ups to record undisturbed ungulate behavior and fresh wolf (Canis lupus) scats as olfactory cue. Habitat visibility at fixed locations in deciduous old growth forest affected neither vigilance levels nor visitation rate and cumulative visitation time of both ungulate species. However, red deer showed a more than two-fold increase of vigilance level from 22% of the time present on control plots to 46% on experimental plots containing one wolf scat. Higher vigilance came at the expense of time spent foraging, which decreased from 32% to 12% while exposed to the wolf scat. These behavioral changes were most pronounced during the first week of the experiment but continuous monitoring of the plots suggested that they might last for several weeks. Wild boar did not show behavioral responses indicating higher perceived predation risk. Visitation rate and cumulative visitation time were not affected by the presence of a wolf scat in both ungulate species. The current study showed that perceived predation risk in red deer and wild boar is not related to habitat visibility in a dense forest ecosystem. However, olfactory cues of wolves affected foraging behavior of their preferred prey species red deer. We showed that odor of wolves in an ecologically equivalent dose is sufficient to create fine-scale risk factors for red deer.