of the Moravian Karst Protected Landscape Area

Abstract Host traits and phylogeny can determine infection risk by driving pathogen transmission and its ability to infect new hosts. Predicting such risks is critical when designing disease mitigation strategies, and especially as regards wildlife, where intensive management is often advocated or prevented by economic and/or practical reasons. We investigated Pseudogymnoascus [Geomyces] destructans infection, the cause of white-nose syndrome (WNS), in relation to chiropteran ecology, behaviour and phylogenetics. While this fungus has caused devastating declines in North American bat populations, there have been no apparent population changes attributable to the disease in Europe. We screened 276 bats of 15 species from hibernacula in the Czech Republic over 2012 and 2013, and provided histopathological evidence for 11 European species positive for WNS. With the exception of Myotis myotis, the other ten species are all new reports for WNS in Europe. Of these, M. emarginatus, Eptesicus nilssonii, Rhinolophus hipposideros, Barbastella barbastellus and Plecotus auritus are new to the list of P. destructans-infected bat species. While the infected species are all statistically phylogenetically related, WNS affects bats from two suborders. These are ecologically diverse and adopt a wide range of hibernating strategies. Occurrence of WNS in distantly related bat species with diverse ecology suggests that the pathogen may be a generalist and that all bats hibernating within the distribution range of P. destructans may be at risk of infection

Low seasonal variation in greater mouse-eared bat (Myotis myotis) blood parameters.

The greater mouse-eared bat (Myotis myotis) is a flagship species for the protection of hibernation and summer maternity roosts in the Western Palearctic region. A range of pathogenic agents is known to put pressure on populations, including the white-nose syndrome fungus, for which the species shows the highest prevalence and infection intensity of all European bat species. Here, we perform analysis of blood parameters characteristic for the species during its natural annual life cycle in order to establish reference values. Despite sexual dimorphism and some univariate differences, the overall multivariate pattern suggests low seasonal variation with homeostatic mechanisms effectively regulating haematology and blood biochemistry ranges. Overall, the species displayed a high haematocrit and haemoglobin content and high concentration of urea, while blood glucose levels in swarming and hibernating bats ranged from hypo- to normoglycaemic. Unlike blood pH, concentrations of electrolytes were wide ranging. To conclude, baseline data for blood physiology are a useful tool for providing suitable medical care in rescue centres, for studying population health in bats adapting to environmental change, and for understanding bat responses to stressors of conservation and/or zoonotic importance

White-nose syndrome pathology grading in Nearctic and Palearctic bats.

While white-nose syndrome (WNS) has decimated hibernating bat populations in the Nearctic, species from the Palearctic appear to cope better with the fungal skin infection causing WNS. This has encouraged multiple hypotheses on the mechanisms leading to differential survival of species exposed to the same pathogen. To facilitate intercontinental comparisons, we proposed a novel pathogenesis-based grading scheme consistent with WNS diagnosis histopathology criteria. UV light-guided collection was used to obtain single biopsies from Nearctic and Palearctic bat wing membranes non-lethally. The proposed scheme scores eleven grades associated with WNS on histopathology. Given weights reflective of grade severity, the sum of findings from an individual results in weighted cumulative WNS pathology score. The probability of finding fungal skin colonisation and single, multiple or confluent cupping erosions increased with increase in Pseudogymnoascus destructans load. Increasing fungal load mimicked progression of skin infection from epidermal surface colonisation to deep dermal invasion. Similarly, the number of UV-fluorescent lesions increased with increasing weighted cumulative WNS pathology score, demonstrating congruence between WNS-associated tissue damage and extent of UV fluorescence. In a case report, we demonstrated that UV-fluorescence disappears within two weeks of euthermy. Change in fluorescence was coupled with a reduction in weighted cumulative WNS pathology score, whereby both methods lost diagnostic utility. While weighted cumulative WNS pathology scores were greater in the Nearctic than Palearctic, values for Nearctic bats were within the range of those for Palearctic species. Accumulation of wing damage probably influences mortality in affected bats, as demonstrated by a fatal case of Myotis daubentonii with natural WNS infection and healing in Myotis myotis. The proposed semi-quantitative pathology score provided good agreement between experienced raters, showing it to be a powerful and widely applicable tool for defining WNS severity

White-Nose Syndrome Fungus: A Generalist Pathogen of Hibernating Bats

<div><p>Host traits and phylogeny can determine infection risk by driving pathogen transmission and its ability to infect new hosts. Predicting such risks is critical when designing disease mitigation strategies, and especially as regards wildlife, where intensive management is often advocated or prevented by economic and/or practical reasons. We investigated <i>Pseudogymnoascus [Geomyces] destructans</i> infection, the cause of white-nose syndrome (WNS), in relation to chiropteran ecology, behaviour and phylogenetics. While this fungus has caused devastating declines in North American bat populations, there have been no apparent population changes attributable to the disease in Europe. We screened 276 bats of 15 species from hibernacula in the Czech Republic over 2012 and 2013, and provided histopathological evidence for 11 European species positive for WNS. With the exception of <i>Myotis myotis</i>, the other ten species are all new reports for WNS in Europe. Of these, <i>M. emarginatus, Eptesicus nilssonii, Rhinolophus hipposideros, Barbastella barbastellus</i> and <i>Plecotus auritus</i> are new to the list of <i>P. destructans</i>-infected bat species. While the infected species are all statistically phylogenetically related, WNS affects bats from two suborders. These are ecologically diverse and adopt a wide range of hibernating strategies. Occurrence of WNS in distantly related bat species with diverse ecology suggests that the pathogen may be a generalist and that all bats hibernating within the distribution range of <i>P. destructans</i> may be at risk of infection.</p></div

<i>Pseudogymnoascus destructans</i> infection in relation to chiropteran phylogeny and ecological similarity.

a<p> =  species using both types of shelters are also included in the previous categories.</p><p>Phylogenetic signal of explanatory variables on a phylogeny of bats from Europe and North America and of <i>P. destructans</i> infection on both phylogeny and a neighbour-joining tree based on squared Euclidean distances of ecological and behavioural traits of hibernating bat species. Values in bold indicate significant clustering or over-dispersion of <i>P. destructans</i> infection on the tree. Note that all categories of explanatory variables were tested here, but <i>n</i> - 1 dummy variables were included in the PGLS model. <i>N</i>  =  number of species scored positive for the given variable, MPD  =  mean phylogenetic distance, NRI  =  net relatedness index, MNTD  =  mean nearest taxon phylogenetic distance, NTI  =  nearest taxon index.</p

Neighbour-joining tree based on ecological and behavioural traits of bats from Europe and North America (rooted at midpoint).

<p>See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0097224#pone-0097224-g002" target="_blank">Figure 2</a> for a description of the colour scheme.</p

Histopathological skin lesions consistent with white-nose syndrome in ten European bat species.

<p>(A) <i>Myotis emarginatus</i>, (B) <i>Eptesicus nilssonii</i>, (C) <i>Rhinolophus hipposideros</i>, (D) <i>Plecotus auritus</i>, (E) <i>Barbastella barbastellus</i>, (F) <i>M. dasycneme</i>, (G) <i>M. nattereri</i>, (H) <i>M. daubentonii</i>, (I) <i>M. bechsteinii</i>, (J) <i>M. brandtii</i>. The photographs illustrate i) extensive infection of the wing membrane and cup-shaped epidermal erosions (A, E, H, J; long black arrow); ii) cup-like epidermal erosions in the pinna (B; long black arrow), iii) <i>Pseudogymnoascus destructans</i> hyphae obscuring the basement membrane and invading the dermis (A, B, C, E, H; black arrow); iv) a single cupping erosion packed with fungal hyphae in the wing membrane (C, D, G, I; long black arrow); v) colonisation of a hair follicle by <i>P. destructans</i>, fungal hyphae present in the associated sebaceous gland and regional connective tissue (F; black arrow); vi) marked signs of inflammation (B, F, J); and vii) a cellular inflammatory crust that sequesters fungal hyphae (A, J). White arrows within each photograph indicate the interface between epidermis and dermis. Periodic acid-Schiff stain; scale bar  =  50 µm. <i>M. myotis</i> not shown because WNS lesions in this species have already been documented elsewhere <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0097224#pone.0097224-Pikula1" target="_blank">[18]</a>.</p

Boxplot of fitted values from the phylogenetic generalised least squares model of <i>P. destructans</i> infection.

<p>Predictions for infected and non-infected species overlap.</p

Species-specific weighted cumulative white-nose syndrome pathology score (histoSum).

<p>Average sum of weighted qualitative scoring for white-nose syndrome severity grades displayed in Figs <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0180435#pone.0180435.g001" target="_blank">1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0180435#pone.0180435.g002" target="_blank">2</a> (± std. error). Animals with histoSum = 1 not classified as positive for WNS on histopathology are included in the figure. Species sampled on multiple continents are presented separately. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0180435#pone.0180435.t001" target="_blank">Table 1</a> for sample sizes.</p