Green spaces contribute to structural resilience of the gut microbiota in urban mammals.

The gut microbiome of wild animals is subject to various environmental influences, including those associated with human-induced alterations to the environment. We investigated how the gut microbiota of a synurbic rodent species, the striped field mouse (Apodemus agrarius), change in cities of varying sizes, seeking the urban microbiota signature for this species. Fecal samples for analysis were collected from animals living in non-urbanized areas and green spaces of different-sized cities (Poland). Metagenomic 16S rRNA gene sequencing and further bioinformatics analyses were conducted. Significant differences in the composition of gut microbiomes among the studied populations were found. However, the observed changes were dependent on local habitat conditions, without strong evidence of a correlation with the size of the urbanized area. The results suggest that ecological detachment from a more natural, non-urban environment does not automatically lead to the development of an "urban microbiome" model in the studied rodent. The exposure to the natural environment in green spaces may serve as a catalyst for microbiome transformations, providing a previously underestimated contribution to the maintenance of native gut microbial communities in urban mammals.

Interspecies transmission of antimicrobial-resistant bacteria between wild birds and mammals in urban environment.

The transmission of antibiotic-resistant bacteria among wild animal species may hold significant epidemiological implications. However, this issue is seldom explored due to the perceived complexity of these systems, which discourages experimental investigation. To address this knowledge gap, we chose a configuration of birds and mammals coexisting in an urban green area as a research model: the rook Corvus frugilegus and the striped field mouse Apodemus agrarius. The indirect transmission of antimicrobial-resistant bacteria between these species is possible because rodents inhabiting rook colonies frequently come into contact with the birds' faeces and pellets. The study was conducted in two cities in eastern Poland (Central Europe) - Lublin and Chelm. Among 71 Escherichia (E.) coli isolates studied, 19.7% showed resistance to from one to six of the antibiotics tested, with much higher prevalence of antibiotic-resistant bacteria in the birds (32%) than in the rodents (7%). Whole genome sequencing was performed on 10 selected E. coli isolates representing similar resistance phenotypes. The following antimicrobial resistance genes were detected: blaTEM-1b, tet(A), tet(B), aph(6)-Id, aph(3'')-Ib, aadA1, aadA2, catA1, floR, cmlA, sul2, sul3, dfrA14, and dfrA2. Birds from the same city and also from both neighbouring cities shared E. coli bacteria with the same sequence types, whereas isolates detected in birds were not found to have been transferred to the mammalian population, despite close contact. This demonstrates that even intensive exposure to sources of these pathogens does not necessarily lead to effective transmission of antibiotic-resistant E. coli strains between birds and mammals. Further efforts should be dedicated to investigating actual transmission of antimicrobial-resistant bacteria in various ecological systems, including those that are crucial for public health, such as urban environments. This will facilitate the development of more accurate models for epidemiological threats and the formulation of well-balanced decisions regarding the coexistence of humans and urban wildlife.

Concentration of metals and metalloids in livers of birds of various foraging guilds collected during the autumn migration period in Poland.

During migration, birds explore various habitats at stopover sites that differ in food resources and contamination levels. In this study, hepatic concentrations of 21 elements (metals and metalloids) in 11 species of birds, representing various foraging habitats (such as aquatic, aquatic/terrestrial, and terrestrial) and migration modes (migratory and sedentary) representing various foraging guilds (omnivores, piscivores, and molluscivores), were analyzed. The samples (N = 84) were collected during the autumn migration period in Poland. The concentrations of elements determined in this study exhibited high inter-species variability, reflecting the diversity in contamination levels depending on food resources used by specific bird groups. Many of the investigated individuals from different species showed exceeded levels of subclinical toxicity and moderate clinical poisoning due to Cd and Hg. Higher concentrations of As, Hg, and Ba and lower V concentrations were found in migratory birds as compared to sedentary birds. Species foraging in terrestrial habitat had different concentrations of some elements compared to aquatic and aquatic/terrestrial species. Some specific inter-species differences in hepatic elemental concentrations were found. Differences in elemental concentrations among various groups can primarily be attributed to their foraging guilds, with certain elements, particularly As, V, and Hg, playing a significant role in the dissimilarity of elemental concentrations between foraging habitat groups and migratory mode groups. The data collected confirmed the limited ability of As to enter ecosystem pathways. The results of this study contribute to understanding the year-round exposure of migratory birds to environmental contamination, which can have carry-over effects on their performance in wintering and breeding grounds.

Lead, cadmium, and other trace elements in the liver of golden eagles and white-tailed eagles: recent data from Poland and a systematic review of previous studies.

The golden eagle (Aquila chrysaetos) and the white-tailed eagle (Haliaeetus albicilla), being apex predators and facultative scavengers, can bioaccumulate different environmental contaminants, including toxic elements that may adversely affect their health. We analyzed the levels of cadmium (Cd), lead (Pb), and other metals and metalloids, including arsenic (As), barium (Ba), beryllium (Be), cobalt (Co), chromium (Cr), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), molybdenum (Mo), selenium (Se), thorium (Th), thallium (Tl), uranium (U), vanadium (V), and zinc (Zn) in liver samples taken from three golden eagles and 36 white-tailed eagles that were found dead across Poland to verify their exposure. We also used a systematic review to summarize the available literature data on Cd, Pb, and other studied elements in the liver of both eagle species. Analyses of trace elements in the liver samples of the Polish eagles revealed interspecific differences in Cd, Cu, and Mn and differences in Co, Mn, Tl, and Zn among study regions. All elements tested except Pb were below the suggested thresholds linked with adverse health effects in birds. The hepatic Pb found in almost half of all the tested individuals suggests environmental exposure to this toxic element. One of the tested white-tailed eagles had hepatic Pb above the threshold of sublethal poisoning. Although our results seem optimistic, as previous Polish studies showed a higher prevalence of birds with hepatic Pb exceeding the toxicity threshold, they indicate that exposure to this toxic metal could still pose an additional threat to the health of Polish eagles.

Identification of Factors Affecting Environmental Contamination Represented by Post-Hatching Eggshells of a Common Colonial Waterbird with Usage of Artificial Neural Networks.

Artificial Neural Networks are used to find the influence of habitat types on the quality of the environment expressed by the concentrations of toxic and harmful elements in avian tissue. The main habitat types were described according to the Corine Land Cover CLC2012 model. Eggs of free-living species of a colonial waterbird, the grey heron Ardea cinerea, were used as a biological data storing media for biomonitoring. For modeling purposes, pollution indices expressing the sum of the concentration of harmful and toxic elements (multi-contamination rank index) and indices for single elements were created. In the case of all the examined indices apart from Cd, the generated topologies were a multi-layer perceptron (MLP) with 1 hidden layer. Interestingly, in the case of Cd, the generated optimal topology was a network with a radial basis function (RBF). The data analysis showed that the increase in environmental pollution was mainly influenced by human industrial activity. The increase in Hg, Cd, and Pb content correlated mainly with the increase in the areas characterized by human activity (industrial, commercial, and transport units) in the vicinity of a grey heron breeding colony. The decrease in the above elements was conditioned by relative areas of farmland and inland waters. Pollution with Fe, Mn, Zn, and As was associated mainly with areas affected by industrial activities. As the location variable did not affect the quality of the obtained networks, it was removed from the models making them more universal.

Dispersal of Aphanoascus keratinophilus by the rook Corvus frugilegus during breeding in East Poland.

The process of dispersal of the potentially disease-causing, geophilic and keratinolytic fungal strain Aphanoascus keratinophilus (the perfect, sexual stage of Chrysosporium keratinophilum) by the rook Corvus frugilegus was studied. The source of A. keratinophilus strains was pellets of the rook, thus far not considered a carrier of this particular opportunistic pathogen. Pellets collected from breeding colonies of rooks were analysed in terms of the occurrence of keratinolytic fungi with the application of the native keratin bait method. Among the 83 rook pellets analysed, 24 (29%) were infected by keratinophilic fungi. Pure cultures of the fungi were identified to species based on traditional morphological features. Traditional mycological identification was verified by the PCR-RFLP molecular identification method as well as DNA sequencing. The obtained results showed the presence of 90 Aphanoascus keratinophilus strains, 6 Chrysosporium tropicum strains, and 3 Chrysosporium pannicola strains. The PCR melting profile (PCR-MP) method was used to identify intraspecies variations of the 90 analysed A. keratinophilus strains. The dispersal of genotypes and possible pathways of A. keratinophilus dispersal and infection via rook pellets were analysed.

Changes in the Montagu's Harrier Circus pygargus diet in Eastern Poland across decades promote insects and reptilians, but not birds and rodents.

We investigated temporal changes in diet composition of the Montagu's Harrier Circus pygargus breeding in natural habitat (calcareous peat bog) in SE Poland. We characterized diet composition in a three-year period (2007-2009), based on pellet analyses. We investigated whether diet composition was affected by years or stage of breeding. We compared diet of the studied population between 2000s and 1990s and with other populations. We found that the food of the studied population was dominated by insects and mammals (by number) and mammals and birds (by biomass). Biomass and abundance of main prey items differed between studied years because of different air temperatures. We found some interannual differences in contribution of some prey items including higher number of thermophilic prey (insects and amphibians) in warmer years. Comparison of pellet composition in the 1990s and 2000s revealed significant increase in the abundance of thermophilic prey (insects and reptiles) and decrease of mammals including Microtus voles and birds. Those changes may be linked to habitat changes in areas neighboring peat bogs and climate change-induced changes in prey communities. The studied population was able to respond to changes in foraging habitats and prey composition by opportunistic foraging on easily available prey. The diet of the studied population is the most similar to the geographically closest populations foraging in similar habitats and characterized by high contribution of insects.

How Is Wildlife Affected by the COVID-19 Pandemic? Lockdown Effect on the Road Mortality of Hedgehogs.

Globally, wildlife is affected by unprecedented changes related to the COVID-19 pandemic. In this paper, the lockdown effect on the traffic-related mortality in hedgehogs in an urban area was studied. Comparing the pre-pandemic (2018 and 2019) and pandemic (2020) years, we showed that hedgehog roadkill levels during the lockdown period were over 50% lower (which means a decrease greater than the decrease in road traffic in the same period measured by the number of accidents or the average number of vehicles per day). Based on literature data, we showed that this may mean at least tens of thousands of hedgehogs have survived on a national scale. We report the need to start intensive research on the possible demographic and genetic effects of this unique phenomenon. We also ask how stable the effect of the COVID-19 pandemic will be on wildlife and whether the lockdown (which is an anthropause) may reverse the negative trends in the decline in the number of wild species, including hedgehogs.

Essential differences in the mineral status of free-ranging European bison Bison bonasus populations in Poland: The effect of the anthroposphere and lithosphere.

Tracking the concentration of elements in the organs of endangered animal species can provide significant knowledge about environmental hazards and allows more targeted protective measures to be taken. In this paper the differences in concentrations of 27 elements in the livers of free-ranging European bison were studied. The ICP-OES method was applied to study the four main Polish populations, which live in lowland (Bialowieska, Knyszynska and Borecka forests) and mountain (Bieszczady) habitats. The study was based on a unique and relatively large amount of comparative material. It was found that European bison populations that live in different locations differ from each other in terms of the hepatic concentrations of 17 of the elements studied in this work. The mountain population in Bieszczady was most distinctive due to the higher concentrations of Ba, Ca, Cd, Se, and the lower concentrations of Mo and V (compared to the other populations). Additionally, even geographically close lowland populations sometimes have different hepatic concentrations of particular elements. The sex and age of individuals have a weak effect on mineral status. Although the lithosphere is the primary factor that affects the availability of elements for the European bison, the soil parameters and biosphere can significantly affect the availability of elements, e.g. by accumulation. It was also shown that the anthroposphere plays a role, but this is mainly through agriculture as feeding on arable fields significantly changes the mineral status of European bison. The need for further research, especially on Cd, Se and Cu, was indicated. It was concluded that toxic elements or deficiencies in essential elements in the populations of the studied species should be considered locally as this would allow better understanding of the health condition of individual populations.

Arthroderma tuberculatum and Arthroderma multifidum Isolated from Soils in Rook (Corvus frugilegus) Colonies as Producers of Keratinolytic Enzymes and Mineral Forms of N and S.

Keratinolytic fungi representing the genus Arthroderma that were isolated from the soils of a rook (Corvus frugilegus) colony were used as biological agents for the disposal of waste feathers. The aim of this study was to assess the abilities of Arthroderma tuberculatum and Arthroderma multifidum fungi with a varied inflow of keratin matter to biodegrade waste feathers. The evaluation was based on the determination of feather mass loss, the activity of keratinolytic enzymes, and the content of mineral N and S forms. It was found that the activity of protease released by the fungi contributed to an increase in the level of soluble proteins and peptides and the concentration of ammonium ions, as well as alkalization of the culture medium. Keratinase activity was significantly correlated with sulfate release, especially in A. tuberculatum cultures. The strains of A. tuberculatum fungi isolated from the soil with the highest supply of organic matter, i.e., strains III, IV, and V, had the lowest enzymatic activity, compared to the A. multifidum strains, but they released mineral nitrogen and sulfur forms that are highly important for fertilization, as well as nutritionally important peptides and amino acids. A. tuberculatum strains can be used for the management of waste feathers that can be applied in agricultural practice.

Inter-species and inter-colony differences in elemental concentrations in eggshells of sympatrically nesting great cormorants Phalacrocorax carbo and grey herons Ardea cinerea.

We compared the concentrations of 17 heavy metals and essential elements in post-hatching eggshells of two waterbirds, the obligate piscivorous great cormorant Phalacrocorax carbo (GCM) and the more omnivorous grey heron Ardea cinerea (GHR), breeding sympatrically in eight mixed colonies in Poland. We found significant inter-species and inter-colony differences in the levels of most of the elements. GHR had significantly higher concentrations of Al, which can be explained by its very low stomach pH: an acidic environment favours the release of Al compounds. Differences in Mn, Ni, Cu, Se and Hg concentrations can be attributed to the various contributions of fish and other aquatic organisms to the diet, and to the exploration of different habitats (GCM exclusively aquatic, GHR a wider range) and microhabitats (GCM, in contrast to wading GHR, dive for food, exploring the whole depth range of water bodies), differently exposed to contamination by those elements from sediments. Inter-colony differences were related to the level of industrialisation. We recorded higher levels of some elements in the eggshells (Fe, Mn in both species and Cr, Ni and Zn in GCM) collected in industrialised areas, which may be associated with the negative environmental impact of industrial areas.

Nests of Marsh harrier (Circus aeruginosus L.) as refuges of potentially phytopathogenic and zoopathogenic fungi.

Birds' nests may be refuges for various species of fungi including that which are potentially phytopathogenic and zoopathogenic. Among the 2449 isolates of fungi obtained from nests of Marsh harriers 96.8% belonged to filamentous fungi. In total, 37 genera were identified from 63 fungi species. Within the mycobiotas of the examined nests populations of fungi which are potentially pathogenic for humans, homoiothermous animals and plants dominated. Among 63 species, 46 (72%) were potentially pathogenic fungi of which 18 species were potentially phytopathogenic and 32 species were pathogenic for homoiothermous animals. Inter alia species of fungi were found in the Marsh harriers nests: Aspergillus fumigatus, Aspergillus flavus, Scopulariopsis brevicaulis, Chrysosporium keratinophilum and Fusarium poae, Fusarium sporotrichioides. In terms of numbers, dominant in Marsh harrier nests were fungi pathogenic to birds, other homoiothermous animals and humans. On that basis it was concluded that Marsh harrier nests are both a source of fungal infections for that species and one of the links in the epidemiological cycle of opportunistic fungi for humans.

Factors Affecting Element Concentrations in Eggshells of Three Sympatrically Nesting Waterbirds in Northern Poland.

Avian eggshells are convenient samples in biomonitoring studies, because they are easily accessible, especially from colonially or semicolonially breeding birds. In the present study, concentrations of 17 elements, including heavy metals and essential elements in post-hatch eggshells, were compared among three species of waterbirds of differing strategies for gaining reserves for egg production and diet: mallard, Anas platyrhynchos (ML, a capital breeder, mainly herbivorous), common tern, Sterna hirundo (CT, an income breeder, piscivorous) and black-headed gull, Chroicocephalus ridibundus (BHG, mixed strategy, omnivorous) and breeding sympatrically in three sites in North Poland. Analyses revealed that Fe, Zn, and Cu levels differed the most in the studied species, which may be explained by various contributions of fish, aquatic plants, and soil invertebrates in their diets. Generally, the studied species' eggshells accumulated amounts of elements comparable to those reported for other waterbirds without putting the growth and development of the embryo at risk. The only exception was very high levels of Cr in ML and CT, which may be explained by their foraging on aquatic organisms in waterbodies polluted by this element. Intersite differences in eggshell concentrations of Ni, Sr, Hg and Cr in CT (an income breeder) may be explained by the influence of local pollution sources (small factories, polluted river).

Trace element concentrations in livers of Common Buzzards Buteo buteo from eastern Poland.

In this study, our aim was to determine the common sources of origin of 18 elements in the livers of Common Buzzards collected during the breeding season in an extensive agricultural landscape in south-east Poland with respect to age (adults and immatures) and sex (males and females). In all 34 specimens collected, the element concentrations followed the pattern of S > Na > Fe > Mg > Zn > Si > Cu > Mn > Ba > Se > B > Pb > Hg > Cd > Cr > Ni > Sr > V. Among the heavy metals examined, only the concentration of Pb was relatively high. Given the prevalence of farmland in the studied area (and the wide use of fertilizers), common use of lead-hunting ammunition and moderate concentration of Pb in fertilizers, the indirect influence of hunting ammunition ingested with food or as gastroliths was apparently responsible for the elevated levels of Pb in the livers of Common Buzzards. In our study, no significant sex-related differences were detected in the hepatic concentrations of any element. However, a significant age effect was observed for three elements, which had elevated levels in adults (Hg) and immature birds (B, Pb), and a significant age x sex interaction was found for S and Fe. These results might be explained by the importance of these elements in bone growth in immature birds (B), variable strategies of foraging between adults and immature birds (Pb), and possible intersex differences in the immature cohort in response to the presence of lead (S, Fe).

Concentrations of lead and other elements in the liver of the white-tailed eagle (Haliaeetus albicilla), a European flagship species, wintering in Eastern Poland.

As a top predator, the white-tailed eagle (Haliaeetus albicilla) may serve as a good indicator species, providing information about the bioavailability of contaminants and their transfer within the food chain. In this study, we aimed to determine the common sources of origin of 17 metals and other elements in the liver of white-tailed eagles, and to compare the variations in their hepatic concentrations by age (adults vs immatures) and sex (males vs females) in groups of white-tailed eagles wintering in Eastern Poland. The element concentrations followed the pattern of S > K > Na > Fe > Mg > Ca > Zn > Cu > Mn > Se > Pb > Hg > Cd > Cr > Sr > V > Sc. We found significant age-related differences in the hepatic concentrations of some of the elements. Adults showed higher concentrations of Pb, Cd, Ca, Fe, and Zn and lower concentrations of Cu and Se than immatures. These differences may be explained by age-related differences in wintering strategy (adults are sedentary, and immatures are migratory) and hunting skills (adults are more successful when hunting for agile prey). Our study indicates that ingested Pb ammunition poses a serious threat to the health and lives of white-tailed eagles in Poland (32% of the studied individuals had acute lead poisoning). Our study also indicates a serious need for banning the use of lead hunting ammunition in the parts of Europe (including Poland) where it is still allowed.

Intra-clutch and inter-colony variability in element concentrations in eggshells of the black-headed gull, Chroicocephalus ridibundus, in northern Poland.

Eggshells are good bioindicators of environmental contamination, and therefore, the concentrations of 17 trace elements in 87 eggshells of black-headed gulls, Chroicocephalus ridibundus, were determined in five breeding colonies in an area dominated by farmland in northern Poland. The intra-clutch variability in the eggshell concentrations of heavy metals and other elements was also investigated, and the concentrations of the elements showed the following pattern: Ca > Mg > Sr > Fe > Zn > Al > Cr > Se > Mn > Cu > Pb > As > Ni > Mo = V > Sc > Cd. The concentrations of Fe, Al, and Mn decreased with the order in which the eggs were laid, but Sr concentrations increased. In contrast, the concentration of Cu significantly increased with the laying date. The concentrations of all elements significantly differed among the studied colonies; the highest concentration of eight elements was found in the eggshells from the Kusowo colony, which may have resulted from the intensive use of fertilizers, manure, and slurry in the surrounding agricultural region. The concentrations of Mg, Sr, and Zn in the eggshells from Skoki Duze were higher than those of the other studied colonies, which may have occurred because the gulls were nesting in a functioning gravel pit; soil and the parent rock are natural reservoirs of these elements. The observed element levels indicate that the environment where the black-headed gull eggs were formed, i.e., primarily near the breeding colonies, remains in a relatively unpolluted state, which was reflected by the low levels of Cd, Ni, and Pb and the lack of measurable levels of Hg.

Influence of landscape features on the location of grey heron Ardea cinerea colonies in Poland.

We analysed hydrographic and habitat factors influencing the location of 207 colonies of the grey heron Ardea cinerea in Poland. We compared areas of particular habitats in three buffers around colonies (0-1, 0-10, 0-20km) among eight regions differing in their proportions of studied habitats. We found the highest inter-region dissimilarities in areas covered by water bodies (all scales) and pastures (0-1km). We recognized some indicator habitats characteristic of the majority (water bodies, pastures) or of some (seacoast, inland marshes, urbanized zone) regions. The habitat selectivity index showed that grey herons in buffer 0-1km preferred pastures and water bodies in seven regions and rivers in one subprovince. In buffer 0-10km, forests and urban zones were preferred in seven and five subprovinces, respectively. Our study revealed that both aquatic and non-aquatic habitats are important for the distribution of the grey heron colonies in Poland.