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Understanding the spatial and temporal patterns of animal road mortality is important for planning protective measures and raising driver awareness. In our study of Lithuania, spanning from 2002 to 2022, we examined these patterns based on road types and categorized them by working days, weekends, and long weekends, predicting the potential impact of transitioning to a four-day working week. We found that road type explains 22–50% of the variability in roadkill numbers for moose, red deer, wild boar, and roe deer. The highest occurrences were on main roads, while regional roads had the fewest incidents. The overall number of roadkills, especially those involving ungulates, was highest on weekends, followed by workdays, with the least on long weekends. However, these variations lacked statistical significance, and their effect size was small. We also observed a trend of increased roadkill numbers on the day before, the first day, notably on All Saints Day, or the last day of long weekends. In this context, with the introduction of a four-day work week, we are only expecting the highest roadkill numbers to move from Friday to Thursday.

We report two new records of occurrence of Six-banded Armadillo, Euphractus sexcinctus (Linnaeus, 1758), in the state of Rondônia, Brazil, based on roadkills. These new records extend the geographic distribution of this species by 70 km. The records are from the region with a high deforestation rate called the “Arc of Deforestation”, which includes the state of Rondônia. Euphractus sexcinctus was recorded in an area with a high loss of vegetation cover, where the forest is being converted to pasture.

‘Animals under wheels’ is a citizen science driven project that has collected almost 90,000 roadkill records from Flanders, Belgium, mainly between 2008 and 2020. However, until now, the platform and results have never been presented comprehensively to the scientific community and we highlight strengths and challenges of this system. Data collection occurred using the subsite www.dierenonderdewielen.be (‘animals under wheels’) or the multi-purpose biodiversity platform observation.org and the apps, allowing the registration of roadkill and living organisms alike. We recorded 4,314 citizen scientists who contributed with at least a single roadkill record (207-1,314 active users per year). Non-roadkill records were registered by 85% of these users and the median time between registration of the first and last record was over 6 years, indicating a very high volunteer retention. Based on photographs presented with the roadkill records (n = 7,687), volunteer users correctly identified 98.2% of the species. Vertebrates represent 99% of all roadkill records. Over 145,000 km of transects were monitored, resulting in 1,726 mammal and 2,041 bird victims. Carcass encounter rates and composition of the top 10 detected species list was dependent on monitoring speed. Roadkill data collected during transects only represented 6% of all roadkill data available in the dataset. The remaining 60,478 bird and mammal roadkill records were opportunistically collected. The top species list, based on the opportunistically collected roadkill data, is clearly biased towards larger, enigmatic species. Although indirect evidence showed an increase in search effort for roadkill from 2010-2020, the number of roadkill records did not increase, indicating that roadkills are diminishing. Mitigation measures preventing roadkill could have had an effect on this, but decrease in population densities was likely to (partially) influence this result. As a case study, the mammal roadkill data were explored. We used linear regressions for the 17 most registered mammal species, determining per species if the relative proportion per year changed significantly between 2010 and 2020 (1 significant decrease, 7 significant increases). We investigated the seasonal patterns in roadkill for the 17 mammal species, and patterns per species were consistent over the years, although restrictions on human movement, due to COVID-19, influenced the seasonal pattern for some species in 2020. In conclusion, citizen scientists are a very valuable asset in investigating wildlife roadkill. While we present the results from Flanders, the platform and apps are freely available for projects anywhere in the world.

The Andean Fox, Lycalopex culpaeus (Molina, 1782), is one of the largest endemic canids in South America. The northernmost part of its distribution lies in the Andes of southwestern Colombia, where it is known only from few localities. We present noteworthy records in southwestern Colombia, including one individual from the south-ern buffer area of Puracé National Park, a second individual found as roadkill on the Pan-American Highway, both in the Department of Cauca, and two additional records from the Colombian Massif. The Puracé record extents the distribution of this species by 74 km northward from previously verified localities.

Background Species of Plasmodium (Haemosporida, Plasmodiidae) are remarkably diverse haemoparasites. Information on genetic diversity of avian malaria pathogens has been accumulating rapidly, however exo-erythrocytic development of these organisms remains insufficiently addressed. This is unfortunate because, contrary to Plasmodium species parasitizing mammals, the avian malaria parasites undergo several cycles of exo-erythrocytic development, often resulting in damage of various organs. Insufficient knowledge on the exo-erythrocytic development in most described Plasmodium species precludes the understanding of mechanisms of virulence during avian malaria. This study extends information on the exo-erythrocytic development of bird malaria parasites. Methods A roadkill fieldfare (Turdus pilaris) was sampled in Switzerland and examined using pathologic, cytologic, histologic, molecular and microbiologic methods. Avian malaria was diagnosed, and erythrocytic and exo-erythrocytic stages of the parasite were identified using morphologic characteristics and barcode DNA sequences of the cytochrome b gene. The species-specific characteristics were described, illustrated, and pathologic changes were reported. Results An infection with Plasmodium matutinum lineage pLINN1 was detected. Parasitaemia was relatively low (0.3%), with all erythrocytic stages (trophozoites, meronts and gametocytes) present in blood films. Most growing erythrocytic meronts were markedly vacuolated, which is a species-specific feature of this parasite's development. Phanerozoites at different stages of maturation were seen in leukocytes, macrophages, and capillary endothelial cells in most organs examined; they were particularly numerous in the brain. Like the erythrocytic meronts, growing phanerozoites were markedly vacuolated. Conspicuous exo-erythrocytic development and maturation in leucocytes suggests that this fieldfare was not adapted to the infection and the parasite was capable to escape from cellular immunity. Conclusions This is the first report of exo-erythrocytic development of the malaria parasite lineage pLINN1 during single infection and the first report of this lineage in the fieldfare. The findings of multiple phanerozoites in brain, skeletal muscle, and eye tissue in combination with signs of vascular blockage and thrombus formation strongly suggest an impaired vision and neuromuscular responsiveness as cause of the unexpected collision with a slowly moving car. Further studies on exo-erythrocytic stages of haemosporidian parasites are pivotal to understand the true level of populational damage of avian malaria in wild birds.

Cervid roadkill, including moose, red deer and roe deer, can pose a risk to drivers and are frequently registered. However, the roadkill influence on overall cervid populations is not fully known, especially by roadkill that are not officially registered. The aim of this study was to assess the impact of cervid roadkill on population abundance, evaluating (i) the proportion of ungulate-vehicle collisions not registered by official bodies, (ii) the number of roadkill in relation to hunted animals and (iii) the proportion of roadkill that occurs in forest habitat. The number of unreported roadkill was calculated based on a roadkill index assessed during 3815 registration sessions on main and national roads in Lithuania from 2002-2022. During this period, 373 moose, 712 red deer and 9179 roe deer roadkill were unreported, correlating to 13.8%, 95.8% and 31.1% of those registered by the Traffic Supervision Service. In conclusion, 39.5%, 17.5% and 20.1% of roadkill were registered on roads through forests. Moose roadkill amounted to a figure corresponding to 10% of those hunted, with the same figure for red deer being 1.8%, neither adding much mortality. At 16.5%-16.6%, the figure for roe deer might be important.

Wildlife-vehicle collisions are an important wildlife conservation challenge, especially for carnivores. In Iran, such vehicle collisions pose a major threat to carnivores. The jungle cat ( Felis chaus ) is a small carnivore species facing multiple threats, including habitat destruction, land use changes, and particularly vehicle collisions. We collected data on jungle cat vehicle collisions in the Hyrcanian forests of northern Iran to model the jungle cat vehicle collision risk and identified high vehicle collision risk areas within the 1 km and 5 km buffers. To check validity of Maxent results, we additionally carried out a binomial Generalized Linear Model (GLM). Results showed that western Golestan province, eastern Mazandaran province, and central Gilan province as the highest vehicle collision risk for the jungle cat in the Hyrcanian forests. According to the Maxent model, human footprint and slope and based on the GLM model, NDVI and human footprint were the most important predictor of jungle cat vehicle collision risk in the Hyrcanian forests. We recommend to implement roadside vegetation management and wildlife crossings in the high collision-risk areas in the Hyrcanian forests to mitigate road mortality and support conservation planning for jungle cats in Iran.

The global dependency on railways as an economical and environmentally‐friendly option for transportation is steadily increasing. Despite their numerous benefits, railways and train traffic can have negative impacts on wildlife, particularly through the risk of mortality due to collisions with trains, entrapment in rails, or electrocution at overhead powerlines. In most cases, these impacts are under‐investigated, thus remain unmitigated. In this study, we describe patterns of rail‐mortality of the local fauna in Balule Nature Reserve, South Africa. In addition to exploring which species are most vulnerable to rail‐mortality, we explore the role that seasonal variation, the daily activity patterns of the species, and the surrounding habitat type have on the occurrence of wildlife‐rail‐mortality. From 25 May 2020–29 March 2021, we conducted carcass surveys over three 5 km railway segments, corresponding to the three dominant habitat types in the nature reserve: open grassland, open woodland and mixed shrubland. Each 5 km segment of railway was surveyed during the wet season (November–March) and the dry season (May–September) to account for seasonal variation. We recorded 99 rail‐kills, of which 29% were mammals, 26% were birds, 26% were reptiles, and 18% were amphibians. Mammal carcasses were found most frequently in the dry season, while amphibians were only detected in the wet season. Amphibian carcasses were all nocturnal species, while diurnal species dominated the bird carcasses found. Finally, most rail‐kill carcasses were found in mixed shrublands, while open woodlands had the lowest frequency of rail‐kill. The detrimental impacts of railway‐mortality on wildlife may be more pronounced in areas of high conservation value, such as nature reserves, and it is essential to study and mitigate these impacts, in order to foster successful co‐existence of wildlife and humans in the landscape.

Trophic dynamics can be altered in complex ways as a result of urbanization. Understanding predator diets in these contexts may not only provide insight into these changes, but also in sources of mortality for vulnerable prey species like the New England cottontail ( Sylvilagus transitionalis ). However, studying the diets of mammalian predators such as bobcats ( Lynx rufus ) can be challenging because of their elusive behavior. DNA metabarcoding of stomach contents from roadkill is a method which provides a new opportunity to study predator diets when mortality events occur. We used this technique to examine variation in bobcat diet across a range of urbanized environments in Connecticut, USA, as well as determine whether bobcats consume the declining New England cottontail. DNA metabarcoding identified between two and five species in the majority of bobcat stomachs. Cottontail ( Sylvilagus spp. ) and eastern gray squirrel ( Sciurus carolinensis ) were each found in over 80% of samples, and most remaining taxa were other small mammals. Nearly a third of the bobcats had consumed white-tailed deer ( Odocoileus virginianus ). Stomach contents containing cottontail remains were sequenced at an additional species-specific marker, but no samples containing the New England cottontail were identified. Bobcats in Connecticut consumed a wide variety of natural prey species including a relatively high proportion of semi-aquatic mammals, and we found no evidence of domestic dog or cat consumption. DNA metabarcoding of stomach contents is an effective approach for opportunistically examining predator diet, and our use of this tool may provide a more complete picture of bobcat diet where other techniques have failed to do so.

Road mortality can be a serious threat to different animals, including snakes. However, mortality patterns can vary between species, intraspecific groups, locations and time. We compared the number of road-killed individuals (carcasses) of two semiaquatic water snakes ( Natrix natrix and N. tessellata ) on 58 km of road sections bordered by an active floodplain and a flood-protected former floodplain on one side and mountainous areas on the other in NE Hungary based on surveys conducted once every two weeks in three non-consecutive years. The results showed high road mortality of snakes, with a spring and an autumn peak corresponding to the times when snakes emerge from and return to hibernating sites. The results show that small-scale spatial differences in road mortality were mediated by landscape structure along the road, while the effects of traffic volume, flood regime and the age and sex of the individuals were negligible. For conservation, the study suggests that establishing culvert passages under the road and/or artificial hibernating sites on the floodplain-side of the roads in critical sections can be promising in reducing road-related mortality.