Occurrence and molecular characterization of Escherichia coli strains isolated from black grouse (Lyrurus tetrix) from the Karkonosze National Park in Poland.

The purpose of this study was to characterize Escherichia coli (E. coli) strains isolated from wild black grouse (Lyrurus tetrix), carried out due to the crossing of hiking trails with wild bird habitats from the Karkonosze National Park. Twenty-seven E. coli isolates were obtained from fecal samples collected during the winter months of 2017 and 2018. The strains were assigned to their relevant phylo-groups and the prevalence of virulence genes characteristic of APEC strains (irp2, astA, iss, iucD, papC, tsh, vat, cva/cvi, stx2f) was checked using PCR analysis. In addition, the phenotypic and genotypic resistance to antibiotics was determined. The entire study provided a better understanding of the potential bacteriological threat to wild birds of the Karkonosze National Park. The results showed that 55.6% of the strains belonged to phylo-group B1 (15/27), 33.3% to group B2 (9/27) and 11.1% to group D (3/27). Among the virulence genes tested, irp2 was detected in 25.9% of isolates (7/27), vat in 22.2% (6/27) and iucD in 3.7% (1/27). The tested E. coli strains showed susceptibility to most antimicrobials, only 14 (51.9%) of them were intermediate resistant or resistant to sulfamethoxazole. The presence of none of the tested genes responsible for resistance to selected antibiotics was identified. Our research indicates a low level of transfer of antimicrobial substances to the natural environment and confirms the effectiveness of the Karkonosze National Park's activities to protect and restore black grouse habitats.

Temperature increase and frost decrease driving upslope elevational range shifts in Alpine grouse and hares.

Global climate change has led to range shifts in plants and animals, thus threatening biodiversity. Latitudinal shifts have been shown to be more pronounced than elevational shifts, implying that northern range edge margins may be more capable to keeping pace with warming than upper elevational limits. Additionally, global climate change is expected to disadvantage habitat specialists. In the Alps, climatic variation along the elevation gradient allows the coexistence of habitat specialists and generalists. Alpine species are anticipated to adapt their elevational ranges to the change of various climate variables caused by global climate change. Regional differences might buffer elevational shifts. Furthermore, distinct climate variables might differently affect the shifts of habitat specialists and generalists. To study the effect of climate change on Alpine species, we analysed hunting bag, climate and biogeographical data of two grouse species (Tetrao tetrix and Lagopus muta) and two hare species (Lepus timidus varronis and L. europaeus) in Grisons, Switzerland, over a period of 30 years. Our results based on 84,630 harvested specimens were as follows: (1) only three out of seven climate variables changed significantly within the study period. (2) The grouse species significantly shifted towards higher elevations, whereas the hare species only shifted in their minimum/maximum elevations. (3) Hunting elevation of habitat generalists increased more than in habitat specialists. (4) The elevational shifts were mostly related to the number of frost days. (5) Hunting elevation increased especially in the southern biogeographical region. To conclude, all four taxa respond to climate change but habitat generalists more rapidly than habitat specialists. The range shift to higher elevations due to global climate change will lead to a reduction in habitat availability for snow-adapted species. Climate change is thus a serious threat to alpine biodiversity. Regions rich in alpine habitats will have an increased responsibility to conserve these species.

Assessing conflicts between winter recreational activities and grouse species.

Mountain environments provide habitats for many wild animal species and are of great importance to recreational activities. In the European Alps, winter recreation activities such as ski mountaineering are becoming increasingly popular, which may lead more often to disturbance of Alpine wildlife. Assessment of ski mountaineering activities and related potential conflict zones is needed to implement protection measures and to guide skiers towards nature-friendly behaviour in sensitive locations. Yet basic data regarding frequencies of ski mountaineers is still missing at regional scale. For the Region of the Tyrol (Austria), this study therefore aimed to advance methods of assessing ski mountaineering activities for use in the spatial analysis of conflict zones with grouse species. We overlaid high-resolution information on the spatial and temporal distribution of ski mountaineers from crowd-sourced data (Strava) with distribution maps of capercaillie (Tetrao urogallus L.) and black grouse (Tetrao tetrix L.). Our results indicate that ski mountaineering activities affected 10.3% of the distribution area of black grouse and 8.6% of the distribution area of capercaillie. The raw crowd-sourced data was pre-processed with local information on skiing resorts, cross-country trails and infrastructure and use of the tested method was validated, with good spatial resolution (0.076 km2 per minimum mapping unit) for large study areas. We identified four intensity classes of ski mountaineering activity, which were randomly distributed in the study area. The spatial distribution of ski mountaineering activities showed an accumulation of activities in the south of the Inn valley. Our results could provide a valuable basis for developing mitigation measures and strategies in order to reduce the disturbance of Alpine wildlife by ski mountaineering.

Molecular analysis of cox-1 and 18S rRNA gene fragments of Eimeria species isolated from endangered grouse: capercaillie (Tetrao urogallus) and black grouse (Tetrao tetrix).

This paper is the first record describing the molecular analysis of Eimeria species occurring in capercaillie (Tetrao urogallus) and black grouse (Tetrao tetrix) which inhabit northern Eurasia and are species critically endangered of extinction. Actions undertaken to protect endangered species, such as breeding individuals in closed aviaries, could allow saving those birds, but they also pose risk of accidental healing of invasive diseases, like coccidiosis. Therefore, an investigation was conducted on fecal samples collected from the capercaillies and black grouse originating from the Kirov region (Russia) and breeding centers located in Poland. Results indicate that the average prevalence of Eimeria revealed 72% (average OPG = 3548) and 80% (average OPG = 5220) in capercaillies and black grouse respectively. Most of the Eimeria spp. oocysts were non-sporulated; however, two different morphological types were observed. The phylogenetic analysis of cox-1 and 18S rRNA genes revealed the analyzed Eimeria sequences to belong to two species. In addition, it showed some similarities between both analyzed genes. Most of the sequences obtained from both grouse species coccidia belonged to one species partially homologous to the Eimeria spp. isolated from ring-necked pheasant (approx. 94 and 96% for cox-1 and 18S rRNA genes, respectively). Two strains isolated from capercaillies imported from Russia were related to turkey coccidia: E. innocua and E. dispersa (97-99% homology) in the cox-1 gene analysis and only one of them was related to those Eimeria species in the 18S rRNA gene analysis (98-99% homology).

Climate change and bird reproduction: warmer springs benefit breeding success in boreal forest grouse.

Global warming is predicted to adversely affect the reproduction of birds, especially in northern latitudes. A recent study in Finland inferred that declining populations of black grouse, Tetrao tetrix, could be attributed to advancement of the time of mating and chicks hatching too early-supporting the mismatch hypothesis. Here, we examine the breeding success of sympatric capercaillie, T. urogallus, and black grouse over a 38-year period in southeast Norway. Breeding season temperatures increased, being most pronounced in April. Although the onset of spring advanced nearly three weeks, the peak of mating advanced only 4-5 days. In contrast to the result of the Finnish study, breeding success increased markedly in both species (capercaillie: 62%, black grouse: 38%). Both brood frequency and brood size increased during the study period, but significantly so only for brood frequency in capercaillie. Whereas the frequency of capercaillie broods was positively affected by rising temperatures, especially during the pre-hatching period, this was not the case in black grouse. Brood size, on the other hand, increased with increasing post-hatching temperatures in both species. Contrary to the prediction that global warming will adversely affect reproduction in boreal forest grouse, our study shows that breeding success was enhanced in warmer springs.

Research Note: Dynamics of black grouse (Tetrao tetrix) infestation with various Eimeria species.

The black grouse (Tetrao tetrix) is an endangered species facing challenges in breeding and reintroduction programs, including parasitic infestations. This study aimed to assess natural infestations by various Eimeria species and infestation dynamics in female, male, and young black grouse kept in a stationary aviary. Faecal samples were collected from adult grouses between April and the time of chicks' hatching and rearing (September). Faecal samples from young birds were collected from the hatching for a period of 1 year. The prevalence of Eimeria spp. was determined by a qualitative method (Fulleborn's flotation) and a quantitative method (McMaster's method with Raynaud's modification). The following Eimeria species were identified: E. lyruri, E. nadsoni and E. nonbrumpti. The average percentages of Eimeria spp. in the cock were 80.52%, 9.27%, and 10.21%, respectively; in the hen, they were 86.19%, 9.28%, and 4.53%, respectively; in the young black grouses they were 84.60%, 9.34% and 6.06%, respectively. The highest E. lyruri infestation was observed in the cock in June (144227 OPG) and July (129365 OPG). In the hen, the infestation intensity increased in May (304302 OPG) and then decreased in June (39583 OPG). Furthermore, an additional increase was observed in July (216533 OPG). Two increases in infestation intensity were also observed in young birds, with peaks in January (91387 OPG) and July (126178 OPG). A positive strong correlation was identified between Eimeria spp. in the cock and the young birds. A statistically significant positive correlation was identified in the hen between E. lyruri and E. nadsoni. No correlation was demonstrated between the infestation intensity and the age of the birds or season of the year in all the grouses under study. Despite some attempts, a comprehensive approach to the issue of coccidiosis in the black grouse as a disease that may affect the success of reintroduction has yet to be established. It seems crucial to monitor the level of Eimeria spp. invasion, and the proposed faecal sampling scheme is an important tool for achieving this goal.

Identifying and locating carotenoids in supra-orbital combs of male black grouse (Lyurus tetrix) using Raman and transmission electron microscopy: A histological study using rehydrated tissue samples.

Colourful signals have long been implicated as indicators of individual quality in animals. Bare-skin signals are an understudied aspect of avian colouration compared with plumage studies, despite displaying rapid changes in size and colour in response to different environmental or physiological stressors. Even fewer studies have focused on the underlying histology of these structures and the importance this plays in the resulting skin colour. Using the Black Grouse (Lyrurus tetrix), we identified the underlying structure of individual dermal spikes, which make up the red supra-orbital comb (a known integumentary signal of male quality), and highlight visual structural differences between combs of different sizes. In addition, we used Raman spectroscopy to indicate the presence of carotenoids within the tissue, something that had previously only been inferred through characteristic reflectance patterns. An increased understanding of the structural basis of colour of featherless parts of the skin opens up exciting new avenues for interpreting the information content of integumentary signals.

Effects of hunting on genetic diversity, inbreeding and dispersal in Finnish black grouse (Lyrurus tetrix).

Intensive hunting activities such as commercial fishing and trophy hunting can have profound influences on natural populations. However, less intensive recreational hunting can also have subtle effects on animal behaviour, habitat use and movement, with implications for population persistence. Lekking species such as the black grouse (Lyrurus tetrix) may be especially prone to hunting as leks are temporally and spatially predictable, making them easy targets. Furthermore, inbreeding in black grouse is mainly avoided through female-biased dispersal, so any disruptions to dispersal caused by hunting could lead to changes in gene flow, increasing the risk of inbreeding. We therefore investigated the impact of hunting on genetic diversity, inbreeding and dispersal on a metapopulation of black grouse in Central Finland. We genotyped 1065 adult males and 813 adult females from twelve lekking sites (six hunted, six unhunted) and 200 unrelated chicks from seven sites (two hunted, five unhunted) at up to thirteen microsatellite loci. Our initial confirmatory analysis of sex-specific fine-scale population structure revealed little genetic structure in the metapopulation. Levels of inbreeding did not differ significantly between hunted and unhunted sites in neither adults nor chicks. However, immigration rates into hunted sites were significantly higher among adults compared to immigration into unhunted sites. We conclude that the influx of migrants into hunted sites may compensate for the loss of harvested individuals, thereby increasing gene flow and mitigating inbreeding. Given the absence of any obvious barriers to gene flow in Central Finland, a spatially heterogeneous matrix of hunted and unhunted regions may be crucial to ensure sustainable harvests into the future.

ENDOPARASITES IN WESTERN CAPERCAILLIES (TETRAO UROGALLUS) AND BLACK GROUSE (TETRAO TETRIX) KEPT IN VARIOUS TYPES OF AVIARIES.

Gallinaceous birds, including Western Capercaillies (Tetrao urogallus) and Black Grouse (Tetrao tetrix), are kept in aviaries and reintroduced to natural habitats as a part of ongoing measures to protect these endangered species. Although parasitic loads can immensely affect reintroduction success of these species, little is known about how the infestation level varies in birds reared with different methods. The aim of this study was to determine the prevalence of endoparasites in fecal samples collected from Galliformes kept in various types of aviaries and transported to reintroduction sites. Most parasitic infections in the examined birds were caused by protozoa of the genus Eimeria, including Eimeria lyruri and Eimeria procera in Western Capercaillies, and E. lyruri, Eimeria nadsoni, and Eimeria tetricis in Black Grouse, which also hosted nematodes of the Capillaria spp. and Ascaridia galli. The prevalence of parasites varied across different types of aviaries. In permanent aviaries, Eimeria spp. was detected in 36% and 35% of the fecal samples collected from Western Capercaillies and Black Grouse, whereas Capillaria spp. were identified in 41% and 69% of the samples, respectively. The prevalence of the identified parasites increased in permanent aviaries on the second sampling date. In contrast, birds kept in seminatural free-flight aviaries in line with the "born to be free" rearing method had a lower prevalence of these parasites. Galliformes housed in seminatural aviaries may be better prepared for reintroduction to their natural habitats.

Gastrointestinal parasites of black grouse Lyrurus tetrix: A long-term study (1986-2019) in the French Alps.

This study investigates the gastroenteric parasites community of 336 black grouse Lyrurus tetrix, covering a wide part of the totality of the species range in the French Alps and characterized by a long-term sampling (1986-2019). Parasite community was described using common epidemiological descriptors. Additionally, for the most prevalent parasite, a species distribution model (SDM) was built to understand the main factors influencing parasite occurrence. Ascaridia compar, Capillaria caudinflata, cestodes (Raillietina urogalli, R.cesticillus, and Hymenolepis microps), Eimeria sp. (E. lyruri and E.nadsoni), Corrigia corrigia and Acanthocephalus sp. The most prevalent species was C.caudinflata (P: 33%,CI95%: 28-38), whereas the lowest prevalence was recorded for Acantocephalus sp. (P:0.3%,CI95%:-0.3-0.9).The selected SDM for C.caudinflata includes only 3 variables: BIO11 (Mean Temperature of Coldest Quarter), the latitude of sampled animals and the distance from ski tour way. The distribution of C.caudinflata in the French Alps presents sharply defined areas where there are optimal environmental conditions for maintaining the life cycle of this parasite. These are the colder areas, characterized by higher latitude values and close to the ski tour ways. This study provides a valuable contribution to the scarce literature on wild Galliformes, describing the composition of the gastrointestinal parasite community of the black grouse.

Changes in the corticosterone level in tooting male black grouse (Tetrao tetrix) infected with Eimeria spp.

The black grouse (Tetrao tetrix) is a forest bird species critically endangered of extinction. Enclosed aviary breeding is among the measures taken to protect the species. Complex factors can affect its reproduction success, including coccidiosis. In this article, corticosterone level (as a reliable biomarker of stress in birds) and prevalence of Eimeria spp. were determined in male black grouse kept in aviary breeding center before, during, and after the reproductive season (called tooting). The correlation between those parameters was also analyzed. The corticosterone level was measured with noninvasive method in fecal samples in an immunoassay. The prevalence of Eimeria spp. was determined by the qualitative (Fülleborn's flotation) and quantitative method (McMaster's method as modified by Raynaud). Results show the occurrence of 3 species of Eimeria spp. in males: Eimeria lyruri, Eimeria nadsoni, and Eimeria nonbrumpti. Infestation with E. lyruri was chronic in nature. A co-invasion of E. lyruri, E. nadsoni, and E. nonbrumpti was observed during (prevalence 60%) and after the tooting (prevalence 40%). The study showed no statistical changes in the oocysts per 1 g of feces (CORTl) and oocysts per 1 g of feces (OPG) in assays I-III and absence of correlation between CORTl and OPG. The results of this experiment do not support the hypothesis that chronic infestation with Eimeria spp. can induce chronic stress in grouse. But may suggest that males of black grouse are susceptible to infestation with other species of Eimeria spp. during mating season. The knowledge of the level of individual stress and parasitic infestation can be used to take protective actions for this bird species, especially to achieve higher survival rate and bird reproduction rate. Tangible effects will include an assessment of the relationship between individual susceptibility and chronic environmental stress caused by coccidia.