A 6-week randomized-controlled field study: effect of isokinetic eccentric resistance training on strength, flexibility and muscle structure of the shoulder external rotators in male junior handball players.

Background: Team handball involves a tremendous amount of shoulder motion with high forces during repeated extended external range of motion. This causes shoulder complaints and overuse injuries. While eccentric training for the lower extremity shows preventive effects by improving strength, range of motion and fascicle length, there is a research gap for the shoulder joint and for advanced tissue characterization using diffusion tensor imaging. Objectives: To investigate the effects of 6-week eccentric isokinetic resistance training on strength, flexibility, and fiber architecture characteristics of the external rotators compared to an active control group in junior male handball players. Methods: 15 subjects were randomly assigned to the eccentric training group and 14 subjects to the active control group (conventional preventive training). Primary outcome measures were eccentric and concentric isokinetic strength of the external rotators, range of motion, and muscle fascicle length and fascicle volume. Results: The intervention group, showed significant changes in eccentric strength (+15%). The supraspinatus and infraspinatus muscles showed significant increases in fascicle length (+13% and +8%), and in fractional anisotropy (+9% and +6%), which were significantly different from the control group. Conclusion: Eccentric isokinetic training has a significant effect on the function and macroscopic structure of the shoulder external rotators in male junior handball players. While strength parameters and muscle structure improved, range of motion did not change. This research helps understanding the physiology of muscle and the role of eccentric training on shoulder function and muscle structure. Furthermore, DTI was found to be a promising tool for advanced tissue characterization, and the in vivo derived data can also serve as model input variables and as a possibility to extend existing ex-vivo muscle models. Future research is needed for functional and structural changes following convenient eccentric field exercises.

Effect of isokinetic eccentric training on the human shoulder strength, flexibility, and muscle architecture in physically active men: A preliminary study.

Strengthening the rotator cuff muscles is important for injury prevention and rehabilitation. Since muscle fascicle length improves motor performance and is suggested to reduce the risk of injury for the hamstring, it may be an important variable to promote multidirectional changes in the function and macroscopic structure for the shoulder. Recent literature reviews overwhelmingly suggest that eccentric exercises improve fascicle length and functional measures for the lower limb. However, there is a research gap for the shoulder. Since ultrasound imaging is the most commonly used imaging technique to quantify muscle structure, but has yielded heterogeneous results in different studies, there is another issue and a research gap for the imaging method. Based on the research gaps, the purpose of this study was to evaluate the effects of standardized eccentric strength training on the function and structure of the external rotator cuff muscles using an isokinetic dynamometer and MRI. Therefore, a preliminary pre-post intervention study was conducted and 16 physically active men were recruited in October 2021. For the right shoulder, an eccentric isokinetic training was performed twice a week for almost six weeks. The primary outcome measures (external rotators) were active and passive range of motion, eccentric and concentric torque at 30, 60, and 180°/s isokinetic speed, and fascicle length and fascicle volume for the supraspinatus and infraspinatus muscles. The findings show a training effect for the absolute mean values of eccentric strength (+24%, p = .008). The torque-angle relationship increased, especially in the final phase of range of motion, although a 4% (p = .002) decrease in passive range of motion was found in the stretch test. Positive changes in muscle structure were shown for the supraspinatus muscle fascicle length (+16%, p = .003) and fascicle volume (+19%, p = .002). Based on the study results, we can conclude that eccentric isokinetic training has a significant positive effect on the shoulder. To our knowledge, this is the first eccentric training study using both isokinetic dynamometer and muscle diffusion tensor imaging to access functional and structural changes in the human shoulder rotator cuff muscles. The methods were shown to be applicable for interventional studies. Based on these results, populations such as high-performance handball players with highly trained shoulders should be included in future studies.

Observational spatial memory in wolves and dogs.

Social learning is highly adaptive in transmitting essential information between individuals in many species. While several mechanisms have been observed, less is known about how much animals can remember. However, results on observational spatial memory among caching species, i.e. a form of social learning allowing individuals to remember and pilfer food caches made by others, suggest that this ability correlates with their social organization. Both wolves and their domesticated form, dogs, are social species known to make food caches, and previous studies have shown that they both can use observational spatial memory abilities to find hidden food. In order to test how much socially transmitted information wolves and dogs can remember, we tested both species in a task requiring them to find 4, 6 or 8 caches after they observed a human hiding food items, or after a control condition where they could not observe the hiding. We found that both wolves and dogs retrieved more caches and were more efficient for the first few caches if they observed the hiding than in the control condition, suggesting that they did not simply rely on scent to find the rewards. Interestingly, wolves outperformed dogs irrespective of whether the caching could be observed or not. We suggest that this result is due to a difference in motivation/persistence between wolves and dogs rather than observational spatial memory.

Diffusion tensor imaging: Influence of segmentation on fiber tracking in the supraspinatus muscle-An inter-operator reliability analysis.

The ability of muscle to generate force depends on its architecture and health condition. MR-based diffusion tensor imaging of muscle (mDTI) is an innovative approach for showing the fiber arrangement for the whole muscle volume. For accurate calculations of fiber metrics, muscle segmentation prior to tractography is regarded as necessary. Since segmentation is known to be operator dependent, it is important to understand how segmentation affects tractography. The aim of this study was to compare the results of deterministic fiber tracking based on muscle models generated by two independent operators. In addition, this study compares the results with a segmentation-free approach. Fifteen subjects underwent mDTI of the right shoulder. The results showed that mDTI can be successfully applied to complex joints such as the human shoulder. Furthermore, operator segmentation did not influence the results of fiber tracking and fascicle length (FL), fiber volume (FV), fractional anisotropy (FA), axial diffusivity (AD), radial diffusivity (RD), and mean diffusivity (MD) showed excellent intraclass correlation estimates (≥ 0.975). As an exploratory approach, the segmentation-free fiber tracking showed significant differences in terms of mean fascicle length. Based on these findings, we conclude that tractography is not sensitive to small deviations in muscle segmentation. Furthermore, it implies that mDTI and automatic segmentation approaches or even a segmentation-free analysis can be considered for evaluation of muscle architecture.

Thermoregulation in the wild boar (Sus scrofa).

The wild boar (Sus scrofa) originates from warm islands but now inhabits large areas of the world, with Antarctica as the only continent not inhabited by this species. One might be tempted to think that its wide distribution results from increasing environmental temperatures. However, any effect of temperature is only indirect: Abundant availability of critical food resources can fully compensate the negative effects of cold winters on population growth. Here, we asked if temperature as a habitat factor is unimportant compared with other habitat indices, simply because wild boars are excellent thermoregulators. We found that the thermoneutral zone in summer was approximately 6-24 °C. In winter, the thermoneutral zone was lowered to 0-7 °C. The estimated increase in the heart rate and energy expenditure in the cold was less than 30% per 10 °C temperature decline. This relatively small increase of energy expenditure during cold exposure places the wild boar in the realm of arctic animals, such as the polar bear, whereas tropical mammals raise their energy expenditure several fold. The response of wild boars to high Ta was weak across all seasons. In the heat, wild boars avoid close contact to conspecifics and particularly use wallowing in mud or other wet substrates to cool and prevent hyperthermia. Wild boars also rely on daily cycles, especially of rhythms in subcutaneous temperature that enables them to cheaply build large core-shell gradients, which serve to lower heat loss. We argue it is predominantly this ability which allowed wild boars to inhabit most climatically diverse areas in the world.

Antimicrobial resistance and its relationship with antimicrobial use on Austrian dairy farms.

The aim of this study was to investigate the prevalence of ESBL/AmpC-producing E. coli and the resistance pattern of commensal E. coli, as well as the link between the use of antibiotics (AMU) and the occurrence of resistance in E. coli on Austrian dairy farms. AMU data from 51 farms were collected over a one-year period in 2020. Fecal samples were collected from cows, pre-weaned and weaned calves in 2020 and 2022. Samples were then analyzed using non-selective and selective agar plates, E. coli isolates were confirmed by MALDI-TOF analysis. Broth microdilution was used for antimicrobial susceptibility testing. The AMU of each farm was quantified as the number of Defined Daily Doses (nDDDvet) and Defined Course Doses (nDCDvet) per cow and year. Cephalosporins (mean 1.049; median 0.732 DDDvet/cow/year) and penicillins (mean 0.667; median 0.383 DDDvet/cow/year) were the most frequently used antibiotics on these farms, followed by tetracyclines (mean 0.275; median 0.084 DDDvet/cow/year). In 2020, 26.8% of the E. coli isolated were resistant to at least one antibiotic class and 17.7% of the isolates were classified as multidrug resistant (≥3 antibiotic classes). Out of 198 E. coli isolates, 7.6% were identified as extended-spectrum/AmpC beta-lactamase (ESBL/AmpC) producing E. coli. In 2022, 33.7% of E. coli isolates showed resistance to at least one antibiotic and 20.0% of isolates displayed multidrug resistance. Furthermore, 29.5% of the samples carried ESBL/AmpC-producing E. coli. In 2020 and 2022, the most frequently determined antibiotic resistances among commensal E. coli isolates were to tetracyclines, sulfonamides and penicillins. In addition, pre-weaned calves had the highest resistance rates in both years. Statistical analyses showed a significant association between low and high use AMU classifications for penicillins (in nDDDvet/cow/year) and their respective resistance among commensal E. coli isolates in 2020 (p = 0.044), as well as for sulfonamide/trimethoprim (p = 0.010) and tetracyclines (p = 0.042). A trend was also noted between the total amount of antibiotics used on farm in 2020 (by nDDDvet/cow/year) and multidrug resistances in commensal E. coli isolated on farm that year (p = 0.067). In conclusion, the relationship between AMU and antimicrobial resistance (AMR) on dairy farms continues to be complex and difficult to quantify.

Sticking Together: Energetic Consequences of Huddling Behavior in Hibernating Juvenile Garden Dormice.

AbstractHibernation, or multiday torpor, allows individuals to save energy via substantial reductions of metabolism and body temperature but is regularly interrupted by euthermic phases called arousals. Social thermoregulation, or "huddling," can act in synergy with torpor in reducing individuals' energy and heat losses. In the wild, the garden dormouse (Eliomys quercinus) combines both strategies, which are crucial for winter survival of juveniles with limited prehibernation body fat reserves. We investigated via thermographic and temperature measurements (i) the energetic impact of huddling during an arousal from deep torpor, (ii) the dynamics of huddling behavior during hibernation, and (iii) its consequences during the entire winter in juvenile garden dormice. Thermographic images revealed a significant effect of huddling on torpor energetics, as it reduced heat exchange and mass loss by two-thirds in huddling versus single individuals during arousal. Our investigation of the dynamics of huddling further revealed a "random-like mechanistic" behavior during winter hibernation, as arousals from torpor were not always initiated by the same individuals. Animals took turns in initiating rewarming within a group, and the individual with highest body temperature during arousal entered into torpor later than the others within the huddle. The animals share both costs and benefits of huddling during arousals, without any energetic benefit of huddling over the entire winter on an individual level. We conclude that the dynamics of social thermoregulation during hibernation seems to counterbalance its benefit of reducing energetic costs associated against the energy-demanding process of rewarming from torpor.

The Effects of Eccentric Strength Training on Flexibility and Strength in Healthy Samples and Laboratory Settings: A Systematic Review.

Background: The risk of future injury appears to be influenced by agonist fascicle length (FL), joint range of motion (ROM) and eccentric strength. Biomechanical observations of the torque-angle-relationship further reveal a strong dependence on these factors. In practice, a longer FL improves sprinting performance and lowers injury risk. Classical stretching is a popular and evidenced-based training for enhancing ROM but does not have any effects on FL and injury risk. However, recent studies show that eccentric-only training (ECC) improves both flexibility and strength, and effectively lowers risk of injury. Objectives: To review the evidence on benefits of ECC for flexibility and strength. Methods: COCHRANE, PUBMED, SCOPUS, SPOLIT, and SPONET were searched for laboratory trials that compare ECC to at least one comparison group. Studies were eligible if they examined both strength and flexibility metrics in a healthy sample (<65 years) and met criteria for controlled or randomized clinical trials (CCT, RCT). 18 studies have been included and successfully rated using the PEDro scale. Results: 16 of 18 studies show strong evidence of strength and flexibility enhancements for the lower limb. While improvements between ECC and concentric training (CONC) were similar for eccentric (+19 ± 10% vs. +19 ± 11%) and isometric strength (+16 ± 10% vs. +13 ± 6%), CONC showed larger improvements for concentric strength (+9 ± 6% vs. +16 ± 7%). While for ROM ECC showed improvements (+9 ± 7%), no results could be found for CONC. The overall effectiveness of ECC seems to be higher than of CONC. Conclusion: There is clear evidence that ECC is an effective method for changes in muscle architecture, leading to both flexibility and strength improvements for the lower limb. Due to limited data no shoulder study could be included. Further research is needed for the upper body joints with a focus on functional and structural adaptions. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42021283248, identifier CRD42021283248.

Cardiovascular effects of intravenous vatinoxan in wild boars (Sus scrofa) anaesthetised with intramuscular medetomidine-tiletamine-zolazepam.

BACKGROUND: The potent sedative medetomidine is a commonly used adjunct for the immobilisation of non-domestic mammals. However, its use is associated with pronounced cardiovascular side effects, such as bradycardia, vasoconstriction and decreased cardiac output. We investigated the effects of the peripherally-acting alpha-2-adrenoceptor antagonist vatinoxan on cardiovascular properties in medetomidine-tiletamine-zolazepam anaesthetised wild boar (Sus scrofa). METHODS: Twelve wild boars, anaesthetised twice with medetomidine (0.1 mg/kg) and tiletamine/zolazepam (2.5 mg/kg) IM in a randomised, crossover study, were administered (0.1 mg/kg) vatinoxan or an equivalent volume of saline IV (control). Cardiovascular variables, including heart rate (HR), mean arterial blood pressure (MAP), pulmonary artery pressure (PAP), pulmonary artery occlusion pressure (PAOP) and cardiac output (CO), were assessed 5 min prior to vatinoxan/saline administration until the end of anaesthesia 30 min later. RESULTS: MAP (p < 0.0001), MPAP (p < 0.001) and MPAOP (p < 0.0001) significantly decreased from baseline after vatinoxan until the end of anaesthesia. HR increased significantly (p < 0.0001) from baseline after vatinoxan administration. However, the effect on HR subsided 3 min after vatinoxan. All variables remained constant after saline injection. There was no significant effect of vatinoxan or saline on CO. CONCLUSION: Vatinoxan significantly reduced systemic and pulmonary artery hypertension, induced by medetomidine in wild boar.

The tarnished silver spoon? Trade-off between prenatal growth and telomere length in wild boar.

Life-history theory predicts a trade-off between growth rates and lifespan, which is reflected by telomere length, a biomarker of somatic state. We investigated the correlation between telomere length and early-life growth of wild boar piglets, Sus scrofa, kept under semi-natural conditions with high food availability to examine our hypothesis that increased pre- and postnatal growth will lead to telomere length attrition, but that a high supply of nutrient may provide the possibility to compensate telomere loss via telomere repair mechanisms. As predicted, our data showed a clear negative correlation between birth body mass and initial telomere length: heavier neonates had shorter telomeres at birth, and we did not find an influence of the mother on initial telomere length. Body mass at birth correlated with body mass later in life and postnatal growth rate did not affect telomere length. We observed an increase in telomere length during postnatal development, suggesting that high food availability allowed piglets to invest into both, growth and telomere restoration. The increase in telomere length over the duration of the study was not accompanied by telomerase activity; thus, telomere elongation was caused either by alternative mechanisms or by short pulses of telomerase activity that we missed. Taken together, this study suggests a trade-off between investment into growth and telomere maintenance even before birth and the possibility to compensate telomere attrition during growth under high amounts of available energy.

Atypical for northern ungulates, energy metabolism is lowest during summer in female wild boars (Sus scrofa).

Typically, large ungulates show a single seasonal peak of heart rate, a proxy of energy expenditure, in early summer. Different to other large ungulates, wild boar females had peak heart rates early in the year (at ~ April, 1), which likely indicates high costs of reproduction. This peak was followed by a trough over summer and a secondary summit in autumn/early winter, which coincided with the mast seeding of oak trees and the mating season. Wild boars counteracted the effects of cold temperatures by decreasing subcutaneous body temperature by peripheral vasoconstriction. They also passively gained solar radiation energy by basking in the sun. However, the shape of the seasonal rhythm in HR indicates that it was apparently not primarily caused by thermoregulatory costs but by the costs of reproduction. Wild boar farrow early in the year, visible in high HRs and sudden changes in intraperitoneal body temperature of females. Arguably, a prerequisite for this early reproduction as well as for high energy metabolism over winter is the broad variety of food consumed by this species, i.e., the omnivorous lifestyle. Extremely warm and dry summers, as experienced during the study years (2017, 2018), may increasingly become a bottleneck for food intake of wild boar.

Dynamic Function and Composition Shift in Circulating Innate Immune Cells in Hibernating Garden Dormice.

Hibernation is characterized by successive torpor bouts during which metabolic rate is down-regulated to 2-4% of euthermic levels along with core body temperatures (T b ) ranging between 0 and 10°C. One characteristic of the torpid state, which is periodically interrupted by a few hours of euthermic phases or arousals during hibernation, resides in an overall impairment of the immune system. The most striking change during torpor is the reduction of circulating white blood cells up to 90%, while their numbers rise to near summer euthermic level upon rewarming. However, potential changes in responsiveness and function of neutrophil granulocytes, accounting for the primary cellular innate immune defense, are unknown. Here we present the first data on shifts in oxidative burst capacity, i.e., the ability to produce reactive oxygen species (ROS), of neutrophils during hibernation. Using a chemiluminescence assay, we measured real-time ROS production in whole blood of hibernating garden dormice (Eliomys quercinus) in early or late torpor, and upon arousals. Accounting for changes in neutrophil numbers along the torpor-arousal cycle, we found significant differences, between torpid and euthermic states, in the neutrophil oxidative burst capacity (NOC), with shallow cell responses during torpor and a highly significant increase by up to 30-fold during arousals. Further, we observed a significant reduction of NOC from aroused animals with euthermic T b of 36.95 ± 0.37°C, when tested at 6°C, whereas no change occurred in NOC from torpid individuals reaching constant T b of 4.67 ± 0.42°C, when measured at 35°C. This dynamic indicates that the reduction in NOC during torpor may be temperature-compensated. These results linked to the understanding of immune function during the torpor-arousal cycle might have clinical relevance in the context of therapeutic hypothermia and reperfusion injury.

Insights in the regulation of trimetylamine N-oxide production using a comparative biomimetic approach suggest a metabolic switch in hibernating bears.

Experimental studies suggest involvement of trimethylamine N-oxide (TMAO) in the aetiology of cardiometabolic diseases and chronic kidney disease (CKD), in part via metabolism of ingested food. Using a comparative biomimetic approach, we have investigated circulating levels of the gut metabolites betaine, choline, and TMAO in human CKD, across animal species as well as during hibernation in two animal species. Betaine, choline, and TMAO levels were associated with renal function in humans and differed significantly across animal species. Free-ranging brown bears showed a distinct regulation pattern with an increase in betaine (422%) and choline (18%) levels during hibernation, but exhibited undetectable levels of TMAO. Free-ranging brown bears had higher betaine, lower choline, and undetectable TMAO levels compared to captive brown bears. Endogenously produced betaine may protect bears and garden dormice during the vulnerable hibernating period. Carnivorous eating habits are linked to TMAO levels in the animal kingdom. Captivity may alter the microbiota and cause a subsequent increase of TMAO production. Since free-ranging bears seems to turn on a metabolic switch that shunts choline to generate betaine instead of TMAO, characterisation and understanding of such an adaptive switch could hold clues for novel treatment options in burden of lifestyle diseases, such as CKD.

How climate change and wildlife management affect population structure in wild boars.

Global climate change affects many species and contributes to the exceptional population growth of wild boar populations and thus to increasing human-wildlife conflicts. To investigate the impact of climate change on wild boar populations we extended existing models on population dynamics. We included for the first time different juvenile conditions to account for long-lasting effects of juvenile body mass on adult body mass and reproductive success. Our analysis shows that incorporating phenotypes, like body mass differences within age classes, has strong effects on projected population growth rates, population structures and the relative importance of certain vital rates. Our models indicated that an increase in winter temperatures and food availability will cause a decrease in mean body mass and litter size within Central European wild boar populations. We further analysed different hunting regimes to identify their effects on the population structure as well as their efficiency in limiting population growth. While targeting juveniles had the lowest effect on population structure, such strategies are, however, rather ineffective. In contrast, culling predominantly yearlings seems very effective. Despite being equally effective, only focusing on adults will not result in a reduction of population size due to their low proportion within populations.

Muscle nonshivering thermogenesis in a feral mammal.

Muscle nonshivering thermogenesis (NST) was recently suggested to play an important role in thermoregulation of species lacking brown adipose tissue (BAT). The mechanism, which is independent of muscle contractions, produces heat based on the activity of an ATPase pump in the sarcoplasmic reticulum (SERCA1a) and is controlled by the protein sarcolipin. To evaluate whether muscle NST could indeed play an important role in thermoregulation in species lacking BAT, we investigated the thermogenic capacities of newborn wild boar piglets. During cold exposure over the first 5 days of life, total heat production was improved while shivering intensity decreased, indicating an increasing contribution of NST. Sampling skeletal muscle tissue for analyses of SERCA activity as well as gene expression of SERCA1a and sarcolipin, we found an age-related increase in all three variables as well as in body temperature. Hence, the improved thermogenesis during the development of wild boars is not due to shivering but explained by the observed increase in SERCA activity. Our results suggest that muscle NST may be the primary mechanism of heat production during cold stress in large mammals lacking BAT, strengthening the hypothesis that muscle NST has likely played an important role in the evolution of endothermy.

Leukocyte Coping Capacity: An Integrative Parameter for Wildlife Welfare Within Conservation Interventions.

Wildlife management, conservation interventions and wildlife research programs often involve capture, manipulation and transport of wild animals. Widespread empirical evidence across various vertebrate taxa shows that handling wildlife generally induces a severe stress response resulting in increased stress levels. The inability of individuals to appropriately respond to rapidly changing environmental conditions during and after manipulations may have deleterious and long-lasting implications on animal welfare. Therefore, mitigating stress responses in the frame of conservation interventions is a key animal welfare factor. However, we have a poor understanding of the metrics to adequately assess and monitor the dynamic physiological changes that animals undergo when subjected to stressful procedures in wild or captive conditions. A growing number of studies provide good evidence for reciprocal interactions between immune processes and stress. Here, we review the existing literature on a relatively new technique-Leukocyte Coping Capacity (LCC), a proxy for stress quantifying oxygen radical production by leukocytes. We discuss the strength and weaknesses of this immunological approach to evaluate stress, the individual capacity to cope with stress and the resulting potential implications for animal welfare. Additionally we present new data on LCC in captive roe deer (Capreolus capreolus) under long-time anesthesia and free-ranging Asiatic wild asses (Kulan; Equus hemionus kulan) were LCC was used to assess stress levels in animals captured for a reintroduction project.

Differences in the reliance on cuticular hydrocarbons as sexual signaling and species discrimination cues in parasitoid wasps.

BACKGROUND: Cuticular hydrocarbons (CHC) have been documented to play crucial roles as species- and sex-specific cues in the chemical communication systems of a wide variety of insects. However, whether they are sufficient by themselves as the sole cue triggering sexual behavior as well as preference of con- over heterospecific mating partners is rarely assessed. We conducted behavioral assays in three representative species of parasitoid wasps (Hymenoptera: Pteromalidae) to determine their reliance on CHC as species-specific sexual signaling cues. RESULTS: We found a surprising degree of either unspecific or insufficient sexual signaling when CHC are singled out as recognition cues. Most strikingly, the cosmopolitan species Nasonia vitripennis, expected to experience enhanced selection pressure to discriminate against other co-occurring parasitoids, did not discriminate against CHC of a partially sympatric species from another genus, Trichomalopsis sarcophagae. Focusing on the latter species, in turn, it became apparent that CHC are even insufficient as the sole cue triggering conspecific sexual behavior, hinting at the requirement of additional, synergistic sexual cues particularly important in this species. Finally, in the phylogenetically and chemically most divergent species Muscidifurax uniraptor, we intriguingly found both CHC-based sexual signaling as well as species discrimination behavior intact although this species is naturally parthenogenetic with sexual reproduction only occurring under laboratory conditions. CONCLUSIONS: Our findings implicate a discrepancy in the reliance on and specificity of CHC as sexual cues in our tested parasitioid wasps. CHC profiles were not sufficient for unambiguous discrimination and preference behavior, as demonstrated by clear cross-attraction between some of our tested wasp genera. Moreover, we could show that only in T. sarcophagae, additional behavioral cues need to be present for triggering natural mating behavior, hinting at an interesting shift in signaling hierarchy in this particular species. This demonstrates the importance of integrating multiple, potentially complementary signaling modalities in future studies for a better understanding of their individual contributions to natural sexual communication behavior.

Seasonal reproductive tactics: annual timing and the capital-to-income breeder continuum.

Tactics of resource use for reproduction are an important feature of life-history strategies. A distinction is made between 'capital' breeders, which finance reproduction using stored energy, and 'income' breeders, which pay for reproduction using concurrent energy intake. In reality, vertebrates use a continuum of capital-to-income tactics, and, for many species, the allocation of capital towards reproduction is a plastic trait. Here, we review how trophic interactions and the timing of life-history events are influenced by tactics of resource use in birds and mammals. We first examine how plasticity in the allocation of capital towards reproduction is linked to phenological flexibility via interactions between endocrine/neuroendocrine control systems and the sensory circuits that detect changes in endogenous state, and environmental cues. We then describe the ecological drivers of reproductive timing in species that vary in the degree to which they finance reproduction using capital. Capital can be used either as a mechanism to facilitate temporal synchrony between energy supply and demand or as a means of lessening the need for synchrony. Within many species, an individual's ability to cope with environmental change may be more tightly linked to plasticity in resource allocation than to absolute position on the capital-to-income breeder continuum.This article is part of the themed issue 'Wild clocks: integrating chronobiology and ecology to understand timekeeping in free-living animals'.

Quantifying capture stress in free ranging European roe deer (Capreolus capreolus).

BACKGROUND: To understand and reduce the concomitant effects of trapping and handling procedures in wildlife species, it is essential to measure their physiological impact. Here, we examined individual variation in stress levels in non-anesthetized European roe deer (Capreolus capreolus), which were captured in box traps and physically restrained for tagging, biometrics and bio-sampling. In winter 2013, we collected venous blood samples from 28 individuals during 28 capture events and evaluated standard measurements for stress (heart rate, body temperature, neutrophil to lymphocyte ratio, lactate and total cortisol). Additionally, we assessed stress using the immunological tool, Leukocyte Coping Capacity (LCC), a real-time proxy for stress measuring oxygen radical production by leukocytes. Finally, the behavioral response to handling was recorded using a scoring system. RESULTS: LCC and therefore stress levels were negatively influenced by the time animals spent in the box trap with human presence at the capture site prior to handling. In contrast, none of the classical stress measures, including total cortisol, nor the behavioral assessment, were correlated with the stressor tested (time of human presence prior to handling) and thus did not provide a clear depiction regarding the extent of the animals short-term stress response. CONCLUSIONS: Overall our study verifies the LCC as a strong method to quantify short-term stress reactions in wildlife. Moreover, our results clearly show that human presence at the trapping site prior to handling should be kept to an absolute minimum in order to reduce stress levels.