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Wildlife in the Anthropocene is increasingly spatially and temporally constrained by lethal and non-lethal human disturbance. For large carnivores with extensive space requirements, like wolves and Eurasian lynx, avoiding human disturbance in European landscapes is challenging when sufficient space with low disturbance is rarely available. Consequently, investigating behavioural adjustments to human presence is critical to understanding the capacity to adapt to human disturbance. We hypothesised that under low human disturbance conditions, large carnivores would adjust their temporal behaviours to make use of daytime, and when daytime human disturbance is high, they would opt for nocturnality. Using camera trap data from nine European study sites along a gradient in human disturbance, we analysed wolf and Eurasian lynx activity patterns. Our data spanned multiple years, 2014 – 2022, and we focused our analysis on September until April, when most large carnivore monitoring takes place. For wolves, our analysis revealed i) increased nocturnal behaviour, ii) decreased diurnal overlap with increasing human activity, and iii) a significant association between a higher probability of nocturnal activity and increasing human disturbance. For Eurasian lynx, we found iv) consistently nocturnal behaviours across all study sites, regardless of human disturbance, and v) no association between human disturbance and increased probability of being active during the night. Our results show that wolves can adjust to diurnal or cathemeral behaviours under low human disturbance, but shift to nocturnality when human disturbance increases. Eurasian lynx, however, consistently maintain their nocturnal behaviour, which we attribute to their principal hunting strategy of stalk and ambush. If human disturbance constrains large carnivore activity to nighttime, it could influence their interactions with prey, leading to cascading effects in the ecosystem. On the other hand, maintaining nocturnal behaviours in human-dominated landscapes may benefit large carnivore conservation, by decreasing negative interactions with humans thereby contributing to a landscape of coexistence.

Abstract The Eurasian lynx is a large carnivore widely distributed across Eurasia. However, our understanding of population status is heterogeneous across their range, with some populations isolated that are at risk of reduced genetic variation and a complete lack of information about others. In many European countries, Eurasian lynx are monitored through demographic studies crucial for their conservation and management. Even so, there are only rough and fragmented population assessments from Ukraine and Belarus, despite strict protection in both countries and their importance for lynx connectivity across Europe. We monitored lynx from October 2020 to March 2021 and used camera trapping in combination with spatial capture–recapture (SCR) methods in a Bayesian framework to provide the first SCR density estimation of three lynx populations across Ukraine and Belarus, including the Ukrainian Chornobyl Exclusion Zone, southern Belarus and the Ukrainian Carpathians. Our density estimates varied within our study areas ranging from 0.45 to 1.54 individuals/100 km2. This work provides a substantial scientific component to the overall understanding of lynx conservation for a region where only broad information is available and opens the doors for further large‐scale monitoring and trend assessments. The crucial information we provide can greatly enhance the range‐wide assessments of the status of this protected species. We also discuss the implications for Eurasian lynx conservation, despite the geopolitical realities impacting species monitoring in the region. Our work serves as a baseline, not only for future conservation interventions but also to evaluate the effects of disturbance and threats to these protected populations.

Alkaloids of the Lycopodiaceae family are of great interest to researchers due to their numerous properties and wide applications in medicine. They play a very important role mainly due to their potent antioxidant, antidepressant effects and a reversible ability to inhibit acetylcholinesterase (AChE) enzyme activity. This property is of immense importance due to the growing problem of an increasing number of patients with neurodegenerative diseases in developed countries and a lack of effective and efficient treatment for them. Numerous studies have shown that Lycopodiaceae alkaloids are a rich source of AChE inhibitors. In the obtaining of new therapeutic phytochemicals from plant material, the extraction process and its efficiency is crucial. Therefore, the aim of this work was to optimize the conditions of modern PLE to obtain bioactive alkaloids from two Lycopodium species: L. clavatum L. and L. annotinum L. Five different solvents of different polarity were used for prepared plant extracts in order to compare the alkaloid content in and thereby effectiveness of the entire extraction. PLE parameters were used based on multiple studies conducted that gave the highest alkaloids recovery. Crude extracts were purified using solid-phase extraction (SPE) on Oasis HLB cartridge and examined by HPLC/ESI-QTOF–MS of the highly abundant alkaloids. To the best of our knowledge, this is the first time such high recoveries have been obtained for known Lycopodiaceae alkaloids. The best extraction results of alkaloid-lycopodine were detected in the dichloromethane extract from L. clavatum, where the yield exceeded 45%. The high recovery of annotinine above 40% presented in L. annotinum was noticed in dichloromethane and ethyl acetate extracts. Moreover, chromatograms were obtained with all isolated alkaloids and the best separation and quality of the bands in methanolic extracts. Interestingly, no alkaloid amounts were detected in cyclohexane extracts belonging to the non-polar solvent. These results could be helpful for understanding and optimizing the best conditions for isolating potent AChE inhibitors.