Pathomorphological Findings and Infectious Diseases in Selected European Brown Hare (Lepus europaeus Pallas, 1778) Populations from Schleswig-Holstein, Germany.

In the northernmost German federal state Schleswig-Holstein, populations of European brown hares (Lepus europaeus Pallas, 1778) show diverse densities and varying courses over the years. To examine differences in pathomorphological findings and infectious diseases as possible reasons for varying population dynamics, we assessed 155 hunted hares from three locations in Schleswig-Holstein from 2016 to 2020. We investigated the association of location, year, age, and sex of animals to certain pathomorphological findings and infectious diseases. Frequent pathomorphological findings were intestinal parasites (63.9%), hepatitis (55.5%), nephritis (31.0%), steatitis (23.2%), enteritis (13.5%), and pneumonia (5.2%). Body condition differed significantly between locations, and the prevalence of pneumonia was significantly higher in females. Enteritis was not detected in 2019, when much more juveniles were sampled. Hepatitis and nephritis occurred significantly more often in 2016 and among adults. Additionally, more adults showed hepatitis with concurrent serotitre for European brown hare syndrome virus (EBHSV), while intestinal parasitosis as well as high excretion rates of coccidia were more common in juveniles. Sampled animals showed high infection rates with Eimeria spp. (96.1%), Trichostrongylus spp. (52.0%), Graphidium strigosum (41.2%), and a high seroprevalence (90.9%) for EBHSV, without severe symptoms. This study revealed a low prevalence of infectious pathogens, but a high prevalence of chronic inflammations of unknown origin in the tested brown hare populations. Overall, our results indicate a rather minor importance of infectious diseases for observed population dynamics of analysed hare populations in Schleswig-Holstein.

Seroprevalences of specific antibodies against avian pathogens in free-ranging ring-necked pheasants (Phasianus colchicus) in Northwestern Germany.

Infectious diseases in captive pheasants (Phasianus colchicus) are well known, but there is a lack of knowledge about occurrence and distribution of pathogens in free-ranging pheasants in Germany. We investigated 604 sera from hunted pheasants and 152 sera from wild caught pheasants between 2011 to 2015, with the aim to determine the prevalence of specific antibodies against different viruses: Avian influenza virus (AIV) of subtypes H5, H7, H9, paramyxovirus type 1 (PMV-1), avian encephalomyelitis virus (AEV), infectious bursitis disease virus (IBDV), infectious bronchitis virus (IBV), infectious laryngotracheitis virus (ILTV), avian metapneumovirus (aMPV) and Salmonella sp., Mycoplasma synoviae (MS) and Mycoplasma gallisepticum (MG). In addition, 178 caeca were investigated for Histomonas meleagridis. The study reveals an ongoing circulation of IBV in the wild pheasant population during the study. Also high seroprevalences of specific antibodies against aMPV depending on the area and a strong increase in prevalence of IBDV antibodies in sera of pheasants in Lower Saxony were detected. ILTV antibody prevalences differed between areas and AEV antibody detection differed between years significantly, whereas specific antibodies against PMV-1 could not be detected and antibodies against AIV-H5, -H7 and -H9 and Mycoplasma spp. were detected in very few cases.

Analysis of hair steroid hormones in polar bears (Ursus maritimus) via liquid chromatography-tandem mass spectrometry: comparison with two immunoassays and application for longitudinal monitoring in zoos.

Analysis of hair cortisol concentrations (HCCs) is a promising method for monitoring long-term stress in mammals. However, previous measurements of HCCs in polar bears (Ursus maritimus) have yielded highly variable results, which are likely due to different methodological approaches. In this study, hair samples of zoo-housed polar bears were analyzed for cortisol with two independent immunoassays [an enzyme-linked immunoassay (EIA) and a chemiluminescence assay (CLIA)] and liquid chromatography-tandem mass spectrometry (LC-MS/MS). HCC measurements depended significantly on assay type applied, sample processing (cutting vs. powdering hair) and their interaction. Best agreement was observed between LC-MS/MS and CLIA (R2 = 0.81 for powdered hair) and sample processing had a minor, albeit significant, effect on obtained HCC measurements in these assays (R2 > 0.9). EIA measurements were consistently higher than with the other assays. HCC measurement was validated biologically for CLIA and LC-MS/MS in one male polar bear that experienced considerable stress for a prolonged period of time (> 18 weeks). Subsequently, by using the validated LC-MS/MS the measurement of cortisol could be complemented by the analysis of other steroids including cortisone, testosterone and progesterone levels from hair samples collected over a 9-month period (5-13 months) from six zoo-housed polar bears (five males, one female). No seasonal steroid variation was observed except in male progesterone levels. For all steroids except cortisone, a strong body region effect (neck or paw) was observed. Cortisol and cortisone, as well as progesterone and testosterone, concentrations were positively correlated. We show that hair steroid concentrations can be used to longitudinally measure stress and reproductive hormone axes in polar bears. The data established herein provide important basic information regarding methodology and study design for assessing hair steroid hormones.

Faecal glucocorticoid metabolites as a measure of adrenocortical activity in polar bears (Ursus maritimus).

Analysis of faecal glucocorticoid metabolites (FGMs) is frequently applied to assess adrenocortical activity in animal conservation and welfare studies. Faecal sample collection is non-invasive and feasible under field conditions. FGM levels are also less prone to circadian rhythms, episodic fluctuations and short acute stressors than glucocorticoid (GC) levels obtained from other matrices, for example blood or saliva. To investigate the suitability of FGM measurement in polar bears (Ursus maritimus), a species listed as Vulnerable by the IUCN (International Union for Conservation of Nature), a cortisol enzyme immunoassay (EIA) was biologically validated by demonstrating a significant increase in FGMs after five zoo-to-zoo transports. In addition to validating the method, the study also documented an average delay of 7 h until the first occurrence of food colorants in the monitored polar bears, which provides essential information for future studies. After validation, the assay was applied to measure FGM concentrations of five polar bears over a 1-year period. Several pre-defined potentially stressful events were recorded in an event log to measure their effect on FGM concentrations. A mixed model analysis revealed significant increases in FGM concentrations after social tension and environmental changes, whereas season and sex had no significant effect. The study demonstrates that the applied cortisol EIA is suitable for measuring FGM levels in polar bears and that using a carefully validated assay for FGM analysis in combination with a detailed sampling protocol can serve as a valuable tool for evaluating mid- to long-term stress in polar bears. FGM levels can be used to monitor stress in captive polar bears in order to optimize housing conditions but also to elucidate stress responses in wild populations for targeted conservation measures.

Effects of multiple exposures to pile driving noise on harbor porpoise hearing during simulated flights-An evaluation tool.

Exploitation of renewable energy from offshore wind farms is substantially increasing worldwide. The majority of wind turbines are bottom mounted, causing high levels of impulsive noise during construction. To prevent temporary threshold shifts (TTS) in harbor porpoise hearing, single strike sound exposure levels (SELSS) are restricted in Germany by law to a maximum of 160 dB re 1 µPa2s at a distance of 750 m from the sound source. Underwater recordings of pile driving strikes, recorded during the construction of an offshore wind farm in the German North Sea, were analyzed. Using a simulation approach, it was tested whether a TTS can still be induced under current protective regulations by multiple exposures. The evaluation tool presented here can be easily adjusted for different sound propagation, acoustic signals, or species and enables one to calculate a minimum deterrence distance. Based on this simulation approach, only the combination of SELSS regulation, previous deterrence, and soft start allow harbor porpoises to avoid a TTS from multiple exposures. However, deterrence efficiency has to be monitored.

Pathological findings in North Sea and Baltic grey seal and harbour seal intestines associated with acanthocephalan infections.

Grey seals Halichoerus grypus and harbour seals Phoca vitulina are common seal species in the North and Baltic seas and final hosts of Corynosoma acanthocephalans. C. strumosum and C. magdaleni infect the small intestines of both seal species. In contrast to harbour seals, Baltic grey seals in the past have regularly displayed severe C. semerme infections in the caecum and colon, with associated tunica muscularis hypertrophy, inflammation and ulcerations as part of the Baltic seal disease complex (BSDC). Pathogenesis and correlation of acanthocephalan infections with these lesions are still unknown. This study describes the intestinal pathology and parasitic distribution in each seal species. Grey seal (n = 83) and harbour seal (n = 1156) intestines of all age groups and sexes, collected in Poland, Germany and Estonia from 1998 to 2017, were investigated. Most harbour seals came from the North Sea, whereas grey seals were predominantly derived from the Baltic Sea. Both species featured mild to moderate small intestinal infections. Grey seals showed colonic infections not found in harbour seals and featured a chronic erosive to ulcerative, eosinophilic or lympho-plasmacytic colitis with tunica muscularis hypertrophy, indicating still prevailing clinical signs of the BSDC. Harbour seals displayed granulomatous, eosinophilic, lympho-plasmacytic or catarrhal enteritis. The prevalence of acanthocephalan infections in harbour seals increased from 2012 onwards. Furthermore, significant associations between acanthocephalan infection and the presence of intestinal inflammation were found for both seal species. This study suggests that the level of acanthocephalan infection and associated lesions are suitable seal population health indicators, with the colon being a specific target organ for Baltic grey seal health monitoring.

Time and tide: Seasonal, diel and tidal rhythms in Wadden Sea Harbour porpoises (Phocoena phocoena).

Odontocetes have evolved a rich diversity of prey- and habitat-specific foraging strategies, which allows them to feed opportunistically on locally and temporally abundant prey. While habitat-specific foraging strategies have been documented for some odontocete species, this is less known for the harbour porpoise (Phocoena phocoena). We collected multiple years of acoustic data using echolocation click loggers to analyse porpoise occurrence and buzzing behaviour, indicating feeding, in the German Wadden Sea (North Sea). Seasonal, diel and tidal effects were studied using Generalised Estimating Equations (GEE-GAMs). Locally season, time of day and tidal time significantly influenced the probability of porpoise detections and detection of foraging sequences (buzzes). Hunting strategies, and therefore frequency of buzzes, were likely affected by prey distribution and large differences between POD locations indicated that porpoises used highly specific behaviour adapted to tide and time of day to efficiently feed on the available prey. Strong seasonal and spatial variation in diel and tidal effects underline the importance of long-term observations. Studies on porpoise behaviour are often based on short-term observations and might rather reflect a seasonal than a general pattern. The results of this study show clearly that significant changes in porpoise behaviour can be found in short and long-term observations. Here some features are based on short term determinants and others are stable over years and care should be taken about drawing general conclusions based on local patterns. Highly variable spatio-temporal patterns indicate a high flexibility of porpoises in a highly variable environment and address a challenge for complex conservation management plans.

The use of seal scarers as a protective mitigation measure can induce hearing impairment in harbour porpoises.

Acoustic deterrent devices (ADDs) are used to deter seals from aquacultures but exposure of harbour porpoises (Phocoena phocoena) occurs as a side-effect. At construction sites, by contrast, ADDs are used to deter harbour porpoises from the zone in which pile driving noise can induce temporary threshold shifts (TTSs). ADDs emit such high pressure levels that there is concern that ADDs themselves may induce a TTS. A harbour porpoise in human care was exposed to an artificial ADD signal with a peak frequency of 14 kHz. A significant TTS was found, measured by auditory evoked potentials, with an onset of 142 dB re 1 µPa2s at 20 kHz and 147 dB re 1 µPa2s at 28 kHz. The authors therefore strongly recommend to gradually increase and down regulate source levels of ADDs to the desired deterrence range. However, further research is needed to develop a reliable relationship between received levels and deterrence.

Selective Predation of a Stalking Predator on Ungulate Prey.

Prey selection is a key factor shaping animal populations and evolutionary dynamics. An optimal forager should target prey that offers the highest benefits in terms of energy content at the lowest costs. Predators are therefore expected to select for prey of optimal size. Stalking predators do not pursue their prey long, which may lead to a more random choice of prey individuals. Due to difficulties in assessing the composition of available prey populations, data on prey selection of stalking carnivores are still scarce. We show how the stalking predator Eurasian lynx (Lynx lynx) selects prey individuals based on species identity, age, sex and individual behaviour. To address the difficulties in assessing prey population structure, we confirm inferred selection patterns by using two independent data sets: (1) data of 387 documented kills of radio-collared lynx were compared to the prey population structure retrieved from systematic camera trapping using Manly's standardized selection ratio alpha and (2) data on 120 radio-collared roe deer were analysed using a Cox proportional hazards model. Among the larger red deer prey, lynx selected against adult males-the largest and potentially most dangerous prey individuals. In roe deer lynx preyed selectively on males and did not select for a specific age class. Activity during high risk periods reduced the risk of falling victim to a lynx attack. Our results suggest that the stalking predator lynx actively selects for size, while prey behaviour induces selection by encounter and stalking success rates.

Patterns of Lynx Predation at the Interface between Protected Areas and Multi-Use Landscapes in Central Europe.

In Central Europe, protected areas are too small to ensure survival of populations of large carnivores. In the surrounding areas, these species are often persecuted due to competition with game hunters. Therefore, understanding how predation intensity varies spatio-temporally across areas with different levels of protection is fundamental. We investigated the predation patterns of Eurasian lynx (Lynx lynx) on roe deer (Capreolus capreolus) and red deer (Cervus elaphus) in both protected areas and multi-use landscapes of the Bohemian Forest Ecosystem. Based on 359 roe and red deer killed by 10 GPS-collared lynx, we calculated the species-specific annual kill rates and tested for effects of season and lynx age, sex and reproductive status. Because roe and red deer in the study area concentrate in unprotected lowlands during winter, we modeled spatial distribution of kills separately for summer and winter and calculated-the probability of a deer killed by lynx and-the expected number of kills for areas with different levels of protection. Significantly more roe deer (46.05-74.71/year/individual lynx) were killed than red deer (1.57-9.63/year/individual lynx), more deer were killed in winter than in summer, and lynx family groups had higher annual kill rates than adult male, single adult female and subadult female lynx. In winter the probability of a deer killed and the expected number of kills were higher outside the most protected part of the study area than inside; in summer, this probability did not differ between areas, and the expected number of kills was slightly larger inside than outside the most protected part of the study area. This indicates that the intensity of lynx predation in the unprotected part of the Bohemian Forest Ecosystem increases in winter, thus mitigation of conflicts in these areas should be included as a priority in the lynx conservation strategy.

Stochastic sampling of interaction partners versus deterministic payoff assignment.

Evolutionary game dynamics describes how successful strategies spread in a population. In well-mixed populations, the usual assumption, e.g. underlying the replicator dynamics, is that individuals obtain a payoff from interactions with a representative sample of the population. This determines their fitness. Here, we analyze a situation in which payoffs are obtained through a single interaction, so that individuals of the same type can have different payoffs. We show analytically that for weak selection, this scenario is identical to the usual approach in which an individual interacts with the whole population. For strong selection, however, differences arise that are reflected in the fixation probabilities and lead to deviating evolutionary dynamics.

Does intra-individual major histocompatibility complex diversity keep a golden mean?

An adaptive immune response is usually initiated only if a major histocompatibility complex (MHC) molecule presents pathogen-derived peptides to T-cells. Every MHC molecule can present only peptides that match its peptide-binding groove. Thus, it seems advantageous for an individual to express many different MHC molecules to be able to resist many different pathogens. However, although MHC genes are the most polymorphic genes of vertebrates, each individual has only a very small subset of the diversity at the population level. This is an evolutionary paradox. We provide an overview of the current data on infection studies and mate-choice experiments and conclude that overall evidence suggests that intermediate intra-individual MHC diversity is optimal. Selective forces that may set an upper limit to intra-individual MHC diversity are discussed. An updated mathematical model based on recent findings on T-cell selection can predict the natural range of intra-individual MHC diversity. Thus, the aim of our review is to evaluate whether the number of MHC alleles usually present in individuals may be optimal to balance the advantages of presenting an increased range of peptides versus the disadvantages of an increased loss of T-cells.