Behavioral Coding of Captive African Elephants (Loxodonta africana): Utilizing DeepLabCut and Create ML for Nocturnal Activity Tracking.

This study investigates the possibility of using machine learning models created in DeepLabCut and Create ML to automate aspects of behavioral coding and aid in behavioral analysis. Two models with different capabilities and complexities were constructed and compared to a manually observed control period. The accuracy of the models was assessed by comparison with manually scoring, before being applied to seven nights of footage of the nocturnal behavior of two African elephants (Loxodonta africana). The resulting data were used to draw conclusions regarding behavioral differences between the two elephants and between individually observed nights, thus proving that such models can aid researchers in behavioral analysis. The models were capable of tracking simple behaviors with high accuracy, but had certain limitations regarding detection of complex behaviors, such as the stereotyped behavior sway, and displayed confusion when deciding between visually similar behaviors. Further expansion of such models may be desired to create a more capable aid with the possibility of automating behavioral coding.

The Effects of Egg- and Substrate-Associated Microbiota on the Larval Performance of the Housefly, Musca domestica.

Increasing human population size and income growth are causing an increasing demand for food and feed. Insects are a more sustainable alternative to conventional animal source proteins, as they can convert waste and by-products from the agricultural industry into biomass for commercial feed for livestock and, potentially, serve as a food source for human consumption. Moreover, insects together with their microorganisms have been shown to play a pivotal role in the development of insects and in the breakdown of complex growth substrates, and are, therefore, closely tied to insect production. This study aims to determine if the removal of egg- and substrate-associated microorganisms impacts larval performance (growth, final biomass, and the survival rate) of M. domestica Linnaeus. Four treatments are tested: disinfected eggs and non-autoclaved substrate, non-disinfected eggs and autoclaved substrate, disinfected eggs and autoclaved substrate, and a control without any removal of microbiota. The results show a significant decrease in the final biomass of larvae subjected to the treatments with only disinfected eggs, only autoclaved substrate, and both compared to the control, and a significant decrease in survival rate for non-disinfected eggs and autoclaved substrate and disinfected eggs and autoclaved substrate compared to the control group. Moreover, larval growth shows a significant difference across days within all treatments. Together, this suggests that the microorganisms of housefly eggs and the growth substrate play an important role in biomass, which is critical in commercial insect production. Together this suggest, that more studies are needed to examine these parameters with respect to more commercially relevant substrates.

Pesticides in the population of European hedgehogs (Erinaceus europaeus) in Denmark.

European hedgehogs (Erinaceus europaeus) inhabit most of Denmark, except for a few smaller islands. Research from other European countries has shown that the hedgehog populations are in decline. The exposure to chemicals might contribute to this development, although their role is currently unknown. Our research studied the occurrence of 19 selected pesticides in the Danish hedgehog population as well as factors potentially explaining the levels of chemicals detected. We analysed 115 liver samples obtained from dead hedgehogs in 2016 for seven rodenticides, four insecticides and eight herbicides commonly used in Denmark at the time of sampling, applying a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method. Detection frequencies varied between 0.9% for fluroxypyr and trans-permethrin and 79% for bromadiolone. Rodenticides, insecticides and herbicides were detected in 84, 43, and 50% of the samples, respectively. The compounds most frequently detected included the insecticide imidacloprid (35%), the herbicide metamitron (29%) and the rodenticide bromadiolone (79%). Individual concentrations varied between non-detected to >2 µg/g. A total of 79% of the 115 hedgehogs contained more than one detectable pesticide, with up to nine of the 19 compounds detected in one individual. The detection frequencies were found to differ significantly between the Eastern and Western part of Denmark for difenacoum, difethialone and imidacloprid. However, no associations were found with sex, age, habitat type or the prevalence of mecC-MRSA and endoparasites in the hedgehogs tested. Whether or not the pesticide levels detected carry a health risk for the hedgehogs remains unknown as no adverse effect levels have yet been established for European hedgehogs for single compounds or pesticide mixtures.

Spatial Distribution and Hierarchical Behaviour of Cattle Using a Virtual Fence System.

Interest in virtual fencing has increased due to its flexibility for agriculture and rewilding. However, systems like Nofence© require large financial investments, and the need for individual collars complicates large-scale use. If cattle herds maintain cohesive groups around leading individuals, fewer collars could be used, thereby enhancing cost efficiency. This study investigates the pattern in spatial distribution in a herd of 17 Angus cows on Fanø in Denmark with GPS locations, using a Nofence© system. The aim of this paper is to determine how individuals position themselves in a herd, spatially, and identify a pattern in ranks. The method used in this study examines the distances between an individual to the rest of the herdmates using nearest neighbour and descriptive statistics. Contrary to expectations, this study did not reveal a distinct pattern in herd distribution. While some tendencies in spatial distribution patterns were observed, only a low concordance could be found (W=0.15,p<0.001), indicating great variability in the cattle's ranks. A cumulative curve of the ranks estimated over the entire periods, however, allowed a rough estimation of the hierarchy and allowed identification of the highest-ranked cows, making the use of a cumulative curve a possible solution to finding the high-ranked cows. This research underscores the complexity of cattle social structures and highlights the need for extended observation periods and alternative methodologies to enhance the cost-effectiveness and scalability of virtual fencing in agricultural and rewilding contexts.

Application of Machine Learning for Automating Behavioral Tracking of Captive Bornean Orangutans (Pongo Pygmaeus).

This article applies object detection to CCTV video material to investigate the potential of using machine learning to automate behavior tracking. This study includes video tapings of two captive Bornean orangutans and their behavior. From a 2 min training video containing the selected behaviors, 334 images were extracted and labeled using Rectlabel. The labeled training material was used to construct an object detection model using Create ML. The use of object detection was shown to have potential for automating tracking, especially of locomotion, whilst filtering out false positives. Potential improvements regarding this tool are addressed, and future implementation should take these into consideration. These improvements include using adequately diverse training material and limiting iterations to avoid overfitting the model.

Strong acclimation effect of temperature and humidity on heat tolerance of the Arctic collembolan Megaphorura arctica.

The Arctic is a highly variable environment in which extreme daily and seasonal temperature fluctuations can occur. With climate change, an increase in the occurrence of extreme high temperatures and drought events is expected. While the effects of cold and dehydration stress on polar arthropods are well studied in combination, little is known about how these species respond to the combined effects of heat and dehydration stress. In this paper, we investigated how the heat tolerance of the Arctic collembola Megaphorura arctica is affected by combinations of different temperature and humidity acclimation regimes under controlled laboratory conditions. The effect of acclimation temperature was complex and highly dependent on both acclimation time and temperature, and was found to have a positive, negative or no effect depending on experimental conditions. Further, we found marked effects of the interaction between temperature and humidity on heat tolerance, with lower humidity severely decreasing heat tolerance when the acclimation temperature was increased. This effect was more pronounced with increasing acclimation time. Lastly, the effect of acclimation on heat tolerance under a fluctuating temperature regime was dependent on acclimation temperature and time, as well as humidity levels. Together, these results show that thermal acclimation alone has moderate or no effect on heat tolerance, but that drought events, likely to be more frequent in the future, in combination with high temperature stress can have large negative impacts on heat tolerance of some Arctic arthropods.

Effects of Social Facilitation and Introduction Methods for Cattle on Virtual Fence Adaptation.

Agricultural industries rely on physical fences to manage livestock. However, these present practical, financial, and ecological challenges, which may be solved using virtual fencing. This study aimed to identify how experienced cattle through social facilitation and the introduction method influence inexperienced cattle. Based on three stocks held in Fanø, Denmark, containing 12, 17 and 13 Angus (Bos taurus), we examined the virtual fence learning in three case studies using one gradual introduction with zero experienced cattle (A) and two different instant introductions with one (B) and ten (C) experienced cattle. Gradual introduction had the virtual fence moved 20 m every other day for eleven days, and in the two instant introductions, the physical fence was removed in one day. Warnings and impulses were recorded during an 11-day learning period and a 26-day post-learning period, using the impulses per warning to quantify if the cattle adapted. Case studies A and B showed a significant reduction in the warnings and impulses, but only A showed a significant reduction in the impulses per warning when comparing the learning period to the post-learning period. Due to the non-standardised experiments, it was not possible to conclude if the number of experienced cattle or the introduction method had an effect on the results.

Using Activity Measures and GNSS Data from a Virtual Fencing System to Assess Habitat Preference and Habitat Utilisation Patterns in Cattle.

There has been an increased focus on new technologies to monitor habitat use and behaviour of cattle to develop a more sustainable livestock grazing system without compromising animal welfare. One of the currently used methods for monitoring cattle behaviour is tri-axial accelerometer data from systems such as virtual fencing technology or bespoke monitoring technology. Collection and transmission of high-frequency accelerometer and GNSS data is a major energy cost, and quickly drains the battery in contemporary virtual fencing systems, making it unsuitable for long-term monitoring. In this paper, we explore the possibility of determining habitat preference and habitat utilisation patterns in cattle using low-frequency activity and location data. We achieve this by (1) calculating habitat selection ratios, (2) determining daily activity patterns, and (3) based on those, inferring grazing and resting sites in a group of cattle wearing virtual fencing collars in a coastal setting with grey, wooded, and decalcified dunes, humid dune slacks, and salt meadows. We found that GNSS data, and a measure of activity, combined with accurate mapping of habitats can be an effective tool in assessing habitat preference. The animals preferred salt meadows over the other habitats, with wooded dunes and humid dune slacks being the least preferred. We were able to identify daily patterns in activity. By comparing general trends in activity levels to the existing literature, and using a Gaussian mixture model, it was possible to infer resting and grazing behaviour in the different habitats. According to our inference of behaviour the herd predominantly used the salt meadows for resting and ruminating. The approach used in this study allowed us to use GNSS location data and activity data and combine it with accurate habitat mapping to assess habitat preference and habitat utilisation patterns, which can be an important tool for guiding management decisions.

A Review of the Occurrence of Metals and Xenobiotics in European Hedgehogs (Erinaceus europaeus).

Monitoring data from several European countries indicate that European hedgehog (Erinaceus europaeus) populations are declining, and research exploring the causes of the decline, including exposure to potentially harmful xenobiotics and metals, may inform conservation initiatives to protect this species in the wild. Hedgehogs are ground-dwelling mammals, feeding on a range of insects, slugs, snails, and earthworms, as well as eggs, live vertebrates, and carrion, including carcasses of apex predator species representing higher levels of the food chain. Consequently, hedgehogs come into close contact with contaminants present in their habitats and prey. This review investigated the studies available on the subject of the occurrence of metals and organic xenobiotics in hedgehogs. This study found that a vast range of different pesticides; persistent organic pollutants (POPs), including organochlorine compounds and brominated flame retardants (BFRs); as well as toxic heavy metals could be detected. Some compounds occurred in lethal concentrations, and some were associated with a potential adverse effect on hedgehog health and survival. Due to their ecology, combined with the opportunity to apply non-invasive sampling techniques using spines as sampling material, we suggest that the European hedgehog is a relevant bioindicator species for monitoring the exposure of terrestrial wildlife to potential toxicants in urban and rural environments.

The possible impact of escaped captive American mink (Neogale vison) on the population of feral mink in Denmark.

The Danish feral mink (Neogale vison) population can be divided into wild-born and captive-born mink newly escaped from a farm. The impact of the captive-born mink on the population of feral population is unknown. Captive-born mink has lately been estimated to make up 25-30% of the feral mink population. In December 2020 Danish mink farms were decommissioned until January 2023. The aim of this study was to: 1) Examine whether a supplementation of farmed mink into the feral mink population influence the population growth and extinction rate of the feral mink, 2) Investigate which parameters affect the viability in feral mink populations the most. The age and mortality of 351 mink and the reproduction of 85 adult females culled by hunters from 2019-2022 were determined in three distinct geographic areas of Denmark (Jutland, Zealand, and Bornholm). VORTEX simulations were used to model the population viability and to simulate the impact of a supplementation of captive-born mink into the wild population. Simulations show that changes by 30% in the parameters: fecundity, percentages of breeding females and mortality influenced population size influenced the final population size significantly. The initial population size and inbreeding did not affect the population growth significantly. The simulations showed that the mink population in the regions of Jutland and Zealand could go extinct within 15 to 30 years without any supplementation of captive-born mink to the feral population. The population on Bornholm will however remain stable with current levels of mortality and reproduction even without supplementation of captive mink.

Computed Tomography as a Method for Age Determination of Carnivora and Odontocetes with Validation from Individuals with Known Age.

Traditional methods for age determination of wildlife include either slicing thin sections off or grinding a tooth, both of which are laborious and invasive. Especially when it comes to ancient and valuable museum samples of rare or extinct species, non-invasive methods are preferable. In this study, X-ray micro-computed tomography (µ-CT) was verified as an alternative non-invasive method for age determination of three species within the order of Carnivora and suborders Odontoceti. Teeth from 13 red foxes (Vulpes vulpes), 2 American mink (Neogale vison), and 2 harbor porpoises (Phocoena phocoena) of known age were studied using µ-CT. The number of visible dental growth layers in the µ-CT were highly correlated with true age for all three species (R2 = 96%, p < 0.001). In addition, the Bland-Altman plot showed high agreement between the age of individuals and visible dental layers represented in 2D slices of the 3D µ-CT images. The true age of individuals was on average 0.3 (±0.6 SD) years higher than the age interpreted by the µ-CT image, and there was a 95% agreement between the true age and the age interpreted from visible dental layers in the µ-CT.

eDNA Metabarcoding- and Microscopic Analysis for Diet Determination in Waterfowl, a Comparative Study in Vejlerne, Denmark.

Understanding diets and structural food webs are keys to the apprehension of ecological communities, upon which conservation and management biology are based. The understanding of grazing and habitat choice for waterfowl is one of the most important topics for avian ecologists today and can, to some degree, be answered by dietary analysis. Droppings collected from four waterfowl, the Eurasian wigeon (Anas penelope), Greylag goose (Anser anser), pink-footed goose (Anser brachyrhynchus) and Barnacle goose (Branta leucopsis) in Vejlerne (Denmark), were analysed microscopically and through eDNA metabarcoding with the use of next generation sequencing (NGS) to accumulate knowledge about the diet of these waterfowl. In total, 120 dropping samples were microscopically analysed, of which the eDNA metabarcoding analysis was done on 79 samples. The prey items were identified according to the taxonomic level of species, and a qualitative method, frequency of occurrence (FO) and FO calculated as a percentage, was used in order to compare the results from the two methods. As neither of the methods was able to encompass all species discovered when combining the two methods, it was concluded in this study that the two methods can support each other in a dietary analysis of waterfowl, but not replace one another.

Revealing the secrets of a 2900-year-old clay brick, discovering a time capsule of ancient DNA.

The recent development of techniques to sequence ancient DNA has provided valuable insights into the civilisations that came before us. However, the full potential of these methods has yet to be realised. We extracted ancient DNA from a recently exposed fracture surface of a clay brick deriving from the palace of king Ashurnasirpal II (883-859 BCE) in Nimrud, Iraq. We detected 34 unique taxonomic groups of plants. With this research we have made the pioneering discovery that ancient DNA, effectively protected from contamination inside a mass of clay, can successfully be extracted from a 2900-year-old clay brick. We encourage future research into this subject, as the scientific prospects for this approach are substantial, potentially leading to a deeper understanding of ancient and lost civilisations.

Effect of Translocation on Host Diet and Parasite Egg Burden: A Study of the European Bison (Bison bonasus).

For the purpose of nature management and species conservation, European bison (Bison bonasus) are being increasingly reintroduced into nature reserves across Europe. The aim of this study was to investigate European bison's adaptability to new areas through the study of their parasite-EPG (eggs per gram feces) and dietary diversity during twelve months after translocation. We compared the parasite-EPG from introduced European bison in Lille Vildmose, Denmark, with the parasite-EPG from populations from Bornholm, Denmark, and Bialowieza Forest, Poland. From March 2021 to February 2022, fecal samples were collected from three populations. Samples from Lille Vildmose were examined through flotation, sedimentation, the Baermann technique, and nanopore sequencing. Fecal samples from Bornholm and Bialowieza were examined through flotation and sedimentation. Nanopore sequencing of DNA from 63 European bison's fecal samples collected during March-September in Lille Vildmose identified 8 species of nematodes within the digestive tract of the European bison, with Haemonchus contortus being the most frequently observed. In Lille Vildmose, a significantly higher excretion of nematode-EPG was observed during the summer period than in the spring, autumn, and winter. In addition, monthly differences in the excretion of nematode eggs were found, with this being significantly higher in June than in the months during autumn and winter (October-February). Significant differences in the nematode-EPG were only found between the excretion of nematode eggs in Bialowieza Forest when compared to that of Lille Vildmose, with significantly higher excretion in Lille Vildmose (October-November). The results indicate that the development rates for nematodes may be affected by changes in temperature, with increasing temperatures speeding up their development time. Independent of this study design, wildlife vets together with the gamekeepers managing the herd found it necessary to treat the herd with antiparasitics for practical and animal welfare reasons in relation to translocation. Furthermore, 79 plant taxa were identified in the diet of the European bison. The broadest diet was observed in March suggesting that the European bison quickly adapted to their new habitat. The results suggest a seasonal shift in their diet, with this being most apparent from March to April.

The Effectiveness of Virtual Fencing of Bull Calves in a Holistic Grazing System.

Large grazers are essential for nature conservation. In order to prevent grazers from moving to unintended areas, it may be necessary to keep them inside enclosures. Physical fences present a number of problems, such as fragmenting the landscape. Virtual fencing, however, is a possible replacement for physical fencing, making it possible to enclose grazers without physical boundaries. Virtual fencing systems utilise collars with GPS technology to track animals and deliver auditory warnings and electric impulses to keep animals within predefined boundaries. This study examines how effective the virtual fencing system Nofence© is at enclosing calves in a holistically managed setting. Holistic management is a rotational grazing technique where an enclosure is grazed in small strips at a time. It is investigated whether the calves become habituated to the virtual fence and whether there is a correlation between the number of warnings received by every two calves in order to explore potential herd behaviour. Finally, this study examines which calves interact the most with the virtual fence by investigating the relationship between physical activity and number of interactions. Seventeen calves were fitted with a GPS collar from the company Nofence© and placed in a holistically managed enclosure. Data were gathered from 4 July to 30 September 2022. The study found that virtual fence was able to contain calves inside the designated enclosure, and over time the calves received notably fewer electrical impulses compared to auditory warnings. The results of Pearson's correlation between auditory warnings received by two random calves were inconclusive, but the use of a sliding window analysis should be further explored. Lastly, the most physically active animals were the ones who received the most auditory warnings, but they did not receive more impulses. No significant correlation was found between the number of electric impulses received and the physical activity of the animals.

Can reintroduction of beavers improve insect biodiversity?

Ecosystem engineering species, such as beavers, may help the restoration of biodiversity. Through the building of dams and lodges and altering the natural hydrology, beavers change the habitat structure and create multiple habitats that facilitate a wide variety of other organisms including terrestrial invertebrate communities. Here we study the effect of beaver reintroduction in Klosterheden in Denmark on biomass of flying invertebrates and diversity of moths. Further, aerial photos were used to assess riparian structure and productivity using the normalized difference vegetation index (NDVI). Our findings show that the presence of beavers affected flying invertebrate biomass, but that this was dependent on time of the year. Further, a strong effect of presence of beavers was found on diversity of moths. The results also show an increase in vegetation productivity and structural heterogeneity at sites with presence of beavers. Overall, our results demonstrate the importance of beavers as important ecosystem engineers that affect invertebrate species composition and abundance, as well as riparian structure and productivity.

Diet of Free Ranging American Mink (Neovison vison) in Denmark.

Non-native American mink (Neovison vison) is a widely spread predator in Denmark. The feral population in mainland Denmark consists of captive-born mink that escaped from fur farms and wild mink born in nature, whereas the population on Bornholm is almost all wild-born mink. In this study, the diets of feral mink in mainland Denmark and on the island of Bornholm are analyzed. The aim of this study was to examine (1) whether the diet of the larger captive-born mink differs from that of the smaller wild-born mink, (2) assess the regional variations between the diets of mink in mainland Denmark and on Bornholm, and (3) investigate the seasonal variation in the diet composition of mink. The stomach contents of 364 mink (243 wild-born and 114 captive-born) culled in the years 2019-2022 were analyzed. Of these, 203 mink were from mainland Denmark, and 154 were from Bornholm. No significant differences were found between the diets of captive-born mink and wild-born mink or the mink found in mainland Denmark and on the island of Bornholm. Significantly more empty stomachs were found during spring than during the summer and autumn, suggesting a bottleneck in the diet during spring.

Selection of Nest Material and Summer Nest Location by the Hazel Dormouse (Muscardinus avellanarius) in the Bidstrup Forests, Denmark.

Hazel dormice (Muscardinus avellanarius) construct summer nests for resting and breeding. The nests are built directly in the vegetation, in tree hollows, or in nest boxes. The availability of nest materials and vegetation coverage may affect the likelihood of finding hazel dormice at a location. The aim of the study is: (1) To investigate the preferences of hazel dormice for nesting materials today compared to four decades ago. (2) To investigate hazel dormice preferences for vegetation coverage at nest sites. In total, 148 hazel dormouse summer nests from the Bidstrup forests in Zealand (Denmark), were analysed. Of these, 82 were collected in the period A: 1980−1985 and 66 were collected in B: 2019−2020. In total 26 different nest materials were found. Beech was the major nest material in both periods, and Jacob's selectivity index indicates that beech is selected for as nesting material and that hazel dormice may travel to collect beech leaves. Nests from period A contained more beech (W = 1521, p < 0.05) and less oak (W = 1304, p < 0.01) compared to nests from period B. Vegetation analysis showed that coverage of shrubs higher than 2 m above ground (W = 1.5, p = 0.07) may be of great importance for hazel dormice.

Testing the Impact of Robotic Lawn Mowers on European Hedgehogs (Erinaceus europaeus) and Designing a Safety Test.

Previous research has established that some models of robotic lawn mowers are potentially harmful to hedgehogs. As the market for robotic lawn mowers is expanding rapidly and the populations of European hedgehogs (Erinaceus europaeus) are in decline, it is important to investigate this risk further to understand the potential threat which some robotic lawn mowers may pose to hedgehogs. We tested 19 models of robotic lawn mowers in collision with hedgehog cadavers to measure their effect on hedgehogs. Our results showed that some models of robotic lawn mowers may injure hedgehogs, whereas others are not harmful to them. Apart from one single incidence, all robotic lawn mowers had to physically touch the hedgehog carcasses to detect them. Larger hedgehog cadavers were less likely to be "injured", with height being the most influential measure of size. The firmness of the tested hedgehog cadavers (frozen or thawed) did not influence the outcome of the collision tests. Neither the position of the hedgehog cadavers nor the selected technical features of the lawn mowers affected the probability of injury. Based on the results, we designed a standardised safety test to measure the effect of a specific model of robotic lawn mower on hedgehogs.

Facing Danger: Exploring Personality and Reactions of European Hedgehogs (Erinaceus europaeus) towards Robotic Lawn Mowers.

The populations of European hedgehog (Erinaceus europaeus) are in decline, and it is essential that research identifies and mitigates the factors causing this. Hedgehogs are increasingly sharing habitats with humans, being exposed to a range of dangers in our backyards. Previous research has documented that some models of robotic lawn mowers can cause harm to hedgehogs. This study explored the personality and behaviour of 50 live hedgehogs when facing an approaching, disarmed robotic lawn mower. By combining a novel arena and novel object test, we found that 27 hedgehogs could be categorised as "shy" and 23 as "bold", independently of sex and age. The encounter tests with a robotic lawn mower showed that the hedgehogs positioned themselves in seven different ways. Personality did not affect their reactions. Adult hedgehogs tended to react in a shyer manner, and the hedgehogs, generally, acted less boldly during their second encounter with the robotic lawn mower. Additionally, our results show that bold individuals reacted in a more unpredictable way, being more behaviourally unstable compared to the shy individuals. This knowledge will be applied in the design of a standardised hedgehog safety test, eventually serving to produce and approve hedgehog-friendly robotic lawn mowers.