Insight into Body Condition Variability in Small Mammals.

The body condition index (BCI) is an indicator of both reproductive success and health in small mammals and might help to understand ecological roles of species. We analyzed BCI data from 28,567 individuals trapped in Lithuania between 1980 and 2023. We compared BCIs between species and examined differences in age groups, gender, and reproductive statuses within each species. Seven out of eighteen species had sample sizes with N < 100. In terms of species, we found that seven of the eight species with the highest average BCIs are granivores or omnivores, which can consume animal-based food at least seasonally. The two contrasting (decreasing or increasing) BCI patterns observed during ontogeny can be related to diet differences among juveniles, subadults, and adult animals. Our results demonstrate that reproductive stress has a negative impact on the BCI of adult females in all analyzed species and nearly all adult males. Although the animals with extremely low BCI consisted mostly of shrews, for the first time we found 23 common and pygmy shrews exhibiting the Chitty effect, i.e., a very high body mass resulting in a BCI > 5.0. This is the first multi-species approach of body condition at middle latitudes. The results increase our understanding of how changing environmental conditions are affecting small mammals.

Trends and Characteristics of Human Casualties in Wildlife-Vehicle Accidents in Lithuania, 2002-2022.

We analyzed 474 human casualties in wildlife-vehicle accidents (WVAs) that occurred between 2002 and 2022 in Lithuania, which is a small northern European country. The study revealed the escalating trend of WVAs, since 2018 surpassing other transport accidents, although the number of casualties per WVA was ca. 100 times lower compared to other transport accidents. Moose was the primary contributor, responsible for 66.7% of fatalities and 47.2% of injuries, despite much lower species abundance compared to roe deer, which is the main species involved in WVAs without human casualties. Temporal patterns highlighted seasonal, daily, and hourly variations, with the majority of casualties occurring during dusk or dawn in May and September, on weekends, and between 20:00 and 22:00. Spatially, main roads with high traffic density exhibited the highest casualties per unit length. Most casualties occurred after hitting an animal directly with cars and motorcycles being most vulnerable vehicles. The effectiveness of WVA prevention measures was inconclusive: 9.5% of fatalities and 1.4% of injuries were registered in the area of the warning sign, and 10.4% of all casualties occurred on fenced road segments. These findings suggest the need for a critical evaluation of the current prevention strategies in reducing human casualties associated with WVAs.

First Observations of Buzzards (Buteo) as Definitive Hosts of Sarcocystis Parasites Forming Cysts in the Brain Tissues of Rodents in Lithuania.

Representatives of the genus Sarcocystis are worldwide distributed apicomplexan parasites characterised by two-host prey-predator relationships. Sarcocystis spp. produce sarcocysts in the muscles and brains of intermediate hosts and develop sporocysts in the intestines of definitive hosts. Two species, Sarcocystis glareoli and Sarcocystis microti, previously assigned to the genus Frenkelia, form cysts in the brains of rodents and are transmitted through the common buzzard (Buteo buteo). In our study, brain samples of 694 small mammals caught in different regions of Lithuania were examined for Sarcocystis spp. Additionally, 10 B. buteo and two rough-legged buzzards (Buteo lagopus) were tested for sporocysts of the analysed parasites. Sarcocystis species were identified based on 28S rRNA sequence comparison. Of the eleven species of small mammals tested, Sarcocystis parasites were observed only in the bank vole (Clethrionomys glareolus). Cysts of S. glareoli were detected in 34 out of 374 C. glareolus (9.1%, 95% CI = 6.4-12.5%). Molecular investigation showed the presence of only S. glareoli in the intestines of 50% of B. buteo. Furthermore, two species, Sarcocystis sp. Rod3 and Sarcocystis sp. Rod4, were confirmed in B. lagopus. Our results demonstrate the need for further studies on Sarcocystis cycling between rodents and birds.

Low Genetic Variability of the Tundra Vole in Lithuania.

The distribution and spread of the tundra vole (Alexandromys oeconomus) in Lithuania have been documented over the last 70 years, but the genetic diversity of the species has not been studied. In this study, we examined A. oeconomus trapped in three sites in northern and western Lithuania using mtDNA sequence analysis of the cytb and control region. The western and northern sites are separated by anthropogenic landscape barriers. The western site is subject to regular spring flooding. Phylogenetic analyses of the studied individuals placed them in the Central European phylogroup, suggesting that Lithuanian A. oeconomus originated from northeastern Poland. In Lithuania, the genetic diversity of A. oeconomus at both mtDNA loci was relatively low (Hd < 0.6, π < 0.002) compared to that found in other European samples (Hd = 0.833-0.958; π = 0.00402-0.01552). Individuals analyzed in Lithuania were genetically different from samples collected in Poland and Northern Europe (ΦST > 0.15, p < 0.05). The genetic divergence between the western and northern samples of A. oeconomus in Lithuania, together with the low genetic variability among the voles studied, provides new insights into the phylogeography of the species and the influence of barriers on the colonization of the country.

Mammal Roadkills in Lithuanian Urban Areas: A 15-Year Study.

We investigated roadkills in urban areas in Lithuania from 2007 to 2022, including two periods with COVID-19 restrictions on people's movement. We analyzed the proportions of wild and domestic animals in roadkill, annual trends, the predominant species involved, and monthly changes during the restrictions. Urban roads were characterized by a low species diversity of roadkilled mammals, with roe deer (Capreolus capreolus) dominating. Total numbers increased exponentially during the study period. The proportion of domestic animals, 12.2%, significantly exceeded that on non-urban roads in the country. The proportion of domestic animals decreased from over 40% in 2007-2009 to 3.7-5.4% in 2020-2022, while the proportion of wild mammals increased from 36.1-39.6% to 89.9-90.6%, respectively. During the periods of COVID-19 restrictions, the number of roadkills in urban areas was significantly higher than expected based on long-term trends. Compared to 2019, the number of roadkilled roe deer in 2020-2021 almost doubled from 700 to 1281-1325 individuals. These anthropause effects were, however, temporary. The imbalance between the roadkill number and transport intensity might require new mitigation strategies to sustain mammal populations in urban areas, at least through improving driver awareness on the issue.

Lockdown's Silver Lining? Different Levels of Roadkill during the COVID-19 Times in Lithuania.

The impact of COVID-19 restrictions on human and vehicular mobility on the number of roadkills of wild mammals, and roe deer in particular, was assessed in Lithuania. We statistically analyzed the distribution of all mammals and roe deer (Capreolus capreolus) as the most abundant species annually from 2002 to 2022 and monthly from 2020 to 2021, including during the two restriction periods. Notably, there was a marked decrease in roadkill incidents on main, national and regional roads, particularly in April-May 2020 (the beginning of lockdown period I) and November-December 2020 (the beginning of lockdown period II), 5.1-20.8% and 33.6-54.4%, respectively. During these months, collisions with mammals on the A14 main road were reduced by 75-90%. However, there was a significant increase in the number of roadkills in urban areas, resulting in the total number of mammal-vehicle and roe deer-vehicle collisions in 2020 and 2021 being higher than expected based on long-term trends. However, after restrictions were eased, collision numbers with wildlife on the main, national and regional roads increased again and became higher than expected.

The extinct Sicilian wolf shows a complex history of isolation and admixture with ancient dogs.

The Sicilian wolf remained isolated in Sicily from the end of the Pleistocene until its extermination in the 1930s-1960s. Given its long-term isolation on the island and distinctive morphology, the genetic origin of the Sicilian wolf remains debated. We sequenced four nuclear genomes and five mitogenomes from the seven existing museum specimens to investigate the Sicilian wolf ancestry, relationships with extant and extinct wolves and dogs, and diversity. Our results show that the Sicilian wolf is most closely related to the Italian wolf but carries ancestry from a lineage related to European Eneolithic and Bronze Age dogs. The average nucleotide diversity of the Sicilian wolf was half of the Italian wolf, with 37-50% of its genome contained in runs of homozygosity. Overall, we show that, by the time it went extinct, the Sicilian wolf had high inbreeding and low-genetic diversity, consistent with a population in an insular environment.

Protozoan Parasites of Sarcocystis spp. in Rodents from Commercial Orchards.

Small mammals are an important group of wildlife that can transmit pathogens to humans and animals. There is a lack of comprehensive studies on the protozoan parasites of the genus Sarcocystis in agricultural areas. The aim of the current research was to evaluate the prevalence of Sarcocystis spp., and to identify the parasite species found in the skeletal muscles of rodents and insectivores from commercial orchards. A total of 679 muscle samples from small mammals, mainly rodents (n = 674), belonging to eight species were examined. Muscle samples were pooled into groups, then digested, and the presence of the Sarcocystis species was confirmed by molecular methods. The examined parasites were determined in five rodent species, Apodemus agrarius, A. flavicollis, Clethrionomys glareolus, Microtus arvalis, and M. oeconomus. The prevalence of Sarcocystis spp. was low: 2.23% in voles and 0.79% in mice. Based on a sequence comparison of cox1 and 28S rDNA, four species were identified: S. myodes, Sarcocystis cf. strixi, Sarcocystis sp. Rod1, and Sarcocystis sp. Rod2. This is the first report of S. myodes in A. agrarius, A. flavicollis, and M. arvalis. The identified species were most closely related to Sarcocystis spp., and were transmitted by predatory mammals and birds. Future studies are needed to describe the species morphologically, as well as to define the host spectrum and to evaluate their possible pathogenicity.

Shrews Under-Represented in Fruit Farms and Homesteads.

Shrews are a less studied group of small mammals than rodents. Between 2018 and 2022, we surveyed 23 sites in Lithuania, including natural and anthropogenic habitats, with the aim to assess the proportion of Soricidae in small mammal communities and their diet based on stable isotope analysis. The average representation of Soricidae was 3.1%, about half the long-term average in other habitats in the country. The highest proportions were in meadows and farmsteads, at 4.9% and 5.0% respectively. Shrews were not trapped on farms or in young orchards, and their relative abundance was very low in intensively managed orchards (0.006 individuals per 100 trap days). Neomys fodiens and N. anomalus were unexpectedly found in homesteads, including in outbuildings. Sorex araneus and S. minutus had similar diets. The trophic carbon/nitrogen discrimination factor between invertebrates and shrew hair was 2.74‱/3.98‱ for S. araneus, 1.90‱/3.78‱ for S. minutus in the orchards. The diet of N. fodiens and N. anomalus at the homesteads requires further investigation. We propose that the under-abundance of shrews may be due to contamination by plant protection products and a lack of invertebrates under intensive agricultural practices.

Abundance and Population Structure of Small Rodents in Fruit and Berry Farms.

Fruit and berry farms are anthropogenic habitats still inhabited by small mammals, though their presence is constantly affected by agricultural activities. Based on trapping data from 2018-2022, we analyzed the abundance and population structure of the dominant rodent species to assess changes in gender and age ratios by year and habitat, the annual and seasonal dynamics of relative abundance, and the relationship between breeding parameters and abundance. The relative abundance of the dominant species, common vole, yellow-necked mouse, striped field mouse, and bank vole, and their proportion in the investigated community varied according to year, season, and habitat. No outbreaks were recorded during the study period. The abundance of the striped field mouse exhibited a downward trend independently of habitat, while the abundance and proportions of the other three species were habitat-dependent. There was no consistent pattern between litter size and relative abundance in the same or following years. Given the ongoing conflict between biodiversity conservation in Europe and agriculture, the results contribute to a better understanding of the functioning and viability of rodent populations in fruit farms and may be used in agroecology and sustainable farming.

The Abundance and Dynamics of Small Mammals and Their Predators: An Editorial.

Small mammals (rodents and insectivores) represent an eclectic group of numerous species of different phylogenetic origins that share similar biological/ecological characteristics derived from their small size [...].

Small Mammal Diversity Changes in a Baltic Country, 1975-2021: A Review.

The structure and diversity of small mammal (SM) communities over the long term may show the influences of climate change, landscape changes and local disturbances. We review published data regarding SM trapping and owl pellet analysis from Lithuania (the most southerly of the three Baltic States, Northern Europe), covering the period 1975-2021. Over decades, we analysed trends in the diversity of SM communities and the proportions of species and proportions of trophic groups. The large increase in granivores, from 6.9% in 1975-1980 to 45.4% in 2011-2020 and 54.7% in 2021, coincided with a decrease in omnivores and insectivores. The proportion of herbivores increased less notably. At the species level, significant decreases in the proportions of M. arvalis, C. glareolus and S. araneus were accompanied by notable increases in the proportions of A. flavicollis and A. agrarius, the latter from 1.0% in 1975-1980 to 25.3% in 2021. Concluding, two periods were identified, specifically before the 1990s and subsequently. In the second period, in the aftermath of land-use changes within the country in 1990, diversity increased, and dominance decreased, a situation that has not subsequently changed. Not excluding the concomitant effects of climate change, we relate these patterns to the alterations in habitat and anthropogenic impact.

Bartonella spp. detection in laelapid (Mesostigmata: Laelapidae) mites collected from small rodents in Lithuania.

The genus Bartonella contains facultative Gram-negative intracellular bacteria from the family Bartonellaceae that can cause diseases in humans and animals. Various Bartonella species have been detected in rodents' ectoparasites, such as fleas, ticks, mites, and lice. However, the role of laelapid mites (Mesostigmata: Laelapidae) as carriers of Bartonella spp. needs to be confirmed. We aimed to investigate the presence of Bartonella spp. in laelapid mites collected from small rodents in Lithuania using real-time PCR targeting the transfer-messenger RNA/tmRNA (ssrA) gene and to characterize Bartonella strains using nested PCR and sequence analysis of the 16S-23S rRNA intergenic transcribed spacer region (ITS). A total of 271 laelapid mites of five species (Laelaps agilis, Haemogamasus nidi, Eulaelaps stabularis, Myonyssus gigas, and Hyperlaelaps microti) were collected from five rodent species (Apodemus flavicollis, Apodemus agrarius, Clethrionomys glareolus, Micromys minutus, and Microtus oeconomus) during 2015-2016. Bartonella DNA was detected in three mite species L. agilis, M. gigas, and Hg. nidi with an overall prevalence of 11.4%. Sequence analysis of the 16S-23S rRNA ITS region revealed the presence of Bartonella taylorii in L. agilis, Hg. nidi, and M. gigas, and Bartonella grahamii in L. agilis. Our results suggest that laelapid mites are involved in the maintenance of rodent-associated Bartonella spp. in nature. To the best of the authors' knowledge, this is the first study to demonstrate the presence of Bartonella spp. DNA in laelapid mites from small rodents.

Factors Affecting Roadkills of Semi-Aquatic Mammals.

We previously showed that registration efforts and traffic intensity explain 90% of variation in mammal roadkill numbers, 70% of variation in the numbers of recorded species and 40% of diversity variation. Here we analyze semiaquatic mammal roadkill in Lithuania in 2002-2021, relating these to the monitoring effort. From 39,936 analyzed roadkill, the European beaver (Castor fiber) was registered 60 times, American mink (Neovison vison) 26 times, otter (Lutra lutra) 22 times and muskrat (Ondatra zibethica) 3 times. The average roadkill indexes were 0.000065, 0.00076, 0.00061 and 0.00010 ind./km/day, and the extrapolated annual roadkill for the country was 44-357, 36-456, 49-464 and 89-144 individuals, respectively. Beaver roadkill numbers correlated with the registration effort and traffic intensity, otter roadkill with registration effort only and mink with hunting bag (number of hunted individuals per year). Roadkill was not always related to proximity to water, with 38-54% of roadkill occurring over 200 m from the nearest water source. With American mink and muskrat being invasive species in the EU and otter protected in many countries, it is valuable to enhance the registrations of their roadkill (using targeted efforts by drivers, hunters or other citizen scientists) to obtain the extrapolated amount of roadkill and to use this knowledge in species management.

Morphological and Molecular Description of Sarcocystis myodes n. sp. from the Bank Vole (Clethrionomys glareolus) in Lithuania

Numerous rodent species have been broadly examined for Sarcocystis parasites. Nevertheless, recent investigations on Sarcocystis spp. in voles are lacking. As many as 45 bank voles (Clethrionomys glareolus) captured in several locations in Lithuania were examined in the present study. Based on morphological, genetic, and phylogenetic results, sarcocysts detected in one bank vole were described as Sarcocystis myodes n. sp. Using light microscopy analysis, the observed sarcocysts were ribbon-shaped, 6000−3000 × 70−220 µm in size. Sarcocysts were characterized by a relatively thin (about 1 µm) and apparently smooth cyst wall. The lancet-shaped bradyzoites were 9.6−12.0 × 3.1−4.6 µm in size. By transmission electron microscopy, the sarcocyst wall was up to 1 µm thick, parasitophorous vacuolar membrane had small knob-like blebs. Based on 18S rDNA, 28S rDNA, cox1, rpoB, and ITS1 loci, S. myodes showed highest similarity with S. ratti from the black rat (Rattus rattus). According to phylogenetic placement, S. myodes was most closely related to Sarcocystis spp. that employ predatory mammals as their definitive hosts. Morphologically, sarcocysts of S. myodes have similar features to those of S. cernae, S. dirumpens, and S. montanaensis described in voles, however, they use birds of prey or snakes as their definitive hosts.

The elemental composition of small mammals in a commercial orchard-meadow system.

We analyze concentrations of various elements in small mammals from commercial orchards and berry plantations and test differences between them depending on species, individual body mass, age, gender, type and age of crop and intensity of applied agricultural measures. Skinned front legs (muscle and bones) were used to register the presence and concentration of Ca, Cd, Mo, Ni, Se, Ag, As, Be, Bi, Co, Cr, Cu, Fe, Ga, Mg, Mn, Pb, Sr, U, V and Zn. The most abundant species were common vole (Microtus arvalis), yellow-necked (Apodemus flavicollis) and striped field (A. agrarius) mice. The maximum recorded concentrations exceeded the minimums by 1.7-7.7 times in Mg, Zn, Cu, Ga, Ni and Ca, and 11.0-23.2 times in Mn, Be, Mo, Co, Sr, V, Pb and As. The hypothesis that the use of fertilization and pesticides in commercial orchards should induce differences in the elemental concentrations between crop areas and control habitats was confirmed by the higher concentrations of Cu, Mn, Bi, Co, Cr, Fe, Ni, Sr and Pb in rodents from the crop areas. Spatially controlled generalized linear mixed model confirmed the cumulative influence of species and crop, explaining 30-80% of the distribution of Ca, Ni, Co, Cu, Ga, Mn, Pb and U. The effect of species and the age of the individual was significant for Ni, Co, Cu, Fe, Ga, Mn and Pb, while effect of gender was not expressed. Depletion of Cu in older individuals was found in all three species. With species as a grouping factor, the effect of crop type and the intensity of agricultural practices were significant factors in the accumulation of Ca, Ni, Co, Cu, Ga, Mn and Pb, while effect of crop age was not expressed. The obtained elemental concentrations in rodents indicated orchards to be cleaner than heavy polluted areas.

Cocirculation of Leptospira spp. and multiple orthohantaviruses in rodents, Lithuania, Northern Europe.

In Europe, zoonotic Leptospira spp. and orthohantaviruses are mainly associated with specific rodent hosts. These pathogens cause febrile human diseases with similar symptoms and disease progression. In Lithuania, the presence of Dobrava-Belgrade orthohantavirus (DOBV), Tula orthohantavirus (TULV) and Leptospira spp. in rodent reservoirs is still unknown, and Puumala orthohantavirus (PUUV) was detected in bank voles (Clethrionomys glareolus) at only one site. Therefore, we collected and screened 1617 rodents and insectivores from Lithuania for zoonotic (re-)emerging Leptospira and orthohantaviruses. We detected Leptospira DNA in six rodent species, namely striped field mouse (Apodemus agrarius), yellow-necked mouse (Apodemus flavicollis), bank vole, common vole (Microtus arvalis), field vole (Microtus agrestis) and root vole (Microtus oeconomus). Leptospira DNA was detected with an overall mean prevalence of 4.4% (range 3.7%-7.9% per rodent species). We detected DOBV RNA in 5.6% of the striped field mice, PUUV RNA in 1% of bank voles and TULV RNA in 4.6% of common voles, but no Leptospira DNA in shrews and no hantavirus-Leptospira coinfections in rodents. Based on the complete coding sequences of the three genome segments, two distant DOBV phylogenetic lineages in striped field mice, one PUUV strain in bank voles and two TULV strains in common voles were identified. The Leptospira prevalence for striped field mice and yellow-necked mice indicated a significant negative effect of the distance to water points. The detection of (re-)emerging human pathogenic Leptospira and three orthohantaviruses in rodent reservoirs in Lithuania calls for increased awareness of public health institutions and allows the improvement of molecular diagnostics for pathogen identification.

Long-term changes in a small mammal community in a temperate zone meadow subject to seasonal floods and habitat transformation.

We tested small mammal (SM) community response to the influence of seasonal floods and habitat transformation in a temporal zone meadow (west Lithuania). The habitat transitioned from a natural flooded meadow in 2008-2012 to a mowed and pastured area in 2020. SM were trapped in late summer to autumn using lines of 25 traps (145 in total), the number of trapping sessions being one or several per year. We tested the hypotheses that: (1) any reaction to the multiple disturbances is species-specific, that is, the relative abundance of SM species and their proportion in the community differs with respect to the flood magnitude and habitat transformation, and (2) at the guild level, disturbances favor habitat generalists, thereby changing the characteristics of SM community. The average relative density of SM was 20.25 individuals per 100 trap nights (range 7.56-40.67), with 4-year-long cyclical changes of density observed. In separate years, we recorded from 4 to 9 species; Shannon's diversity index varied from 0.46 to 2.19, with expressed change of the dominant species. Habitat generalist species were favored, while habitat specialists were disadvantaged. As the meadow transformation progressed, unfavored states in the SM community prevailed, with excessive numbers of granivore and herbivore species present. Our study suggests that multiple disturbances may lead to an increase in relative abundance, species richness, and diversity within the SM assemblages.

Roadkills as a Method to Monitor Raccoon Dog Populations.

The raccoon dog (Nyctereutes procyonoides) is one of the most frequently killed species on Lithuanian roads. As an invasive species, up-to-date knowledge of population size, trends and spatial distribution is critically important both for species assessment and for the planning of control measures. In Lithuania, however, raccoon dog surveys have not been carried out since 1997. We investigated, therefore, whether roadkill counts on predefined routes could be used as a proxy for a survey. Our dataset includes survey numbers for the period 1956-1997, hunting bag sizes for 1965-2020 (including the spatial distribution of the hunting bag in 2018-2020) and roadkill data relating to 1551 individuals between 2002-2020. At the most local scale, that of the hunting areas of hunting clubs, correlations between the numbers of hunted and roadkilled individuals were negative and insignificant or absent. At the country scale, however, we found significant correlation both between the numbers surveyed and hunted in 1965-1997 (r = 0.88), and between those hunted and the number of roadkills in 2002-2020 (r = 0.56-0.69). Therefore, we consider that roadkill counts on predefined and stable routes may be used as a proxy for a survey at the country scale. Practical implementation of the method is proposed.

Isotopic Niche of Syntopic Granivores in Commercial Orchards and Meadows.

In agricultural habitats, diets and trophic positions of syntopic granivorous small mammals are not known sufficiently. Agroecosystems may be quite complex isotopically and the most complex situation concerns the nitrogen-15 isotope as δ15N values are influenced by many internal and external fluxes. We analysed the isotopic niches of striped field (Apodemus agrarius), yellow-necked (Apodemus flavicollis), and harvest (Micromys minutus) mice living sympatrically and syntopically in apple and plum orchards, raspberry and currant plantations, and nearby meadows that were used as control habitats. Carbon (δ13C) and nitrogen (δ15N) stable isotope ratios from hair samples were used as a proxy for their diet. As the main hypothesis, we expected differences in the isotopic niches of these three species, minimising interspecific competition. All species were fully separated according to δ15N values, presuming different amounts of food of animal origin in their diet. The separation of species according to δ13C was not expressed in all habitats. The core dietary niches of these species were fully separated in the apple orchards and raspberry plantations. Intraspecific differences of the isotopic niche were not present in any of the three species: that is, resources were equally used by males and females, adults, subadults, and juveniles.