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.

CEITEC Central European Institute of Technology University of Veterinary and Pharmaceutical Sciences Brno Brno Czech Republic

Department of Botany and Zoology Masaryk University Brno Czech Republic

Department of Ecology and Diseases of Game Fish and Bees University of Veterinary and Pharmaceutical Sciences Brno Brno Czech Republic

Department of Veterinary Pathobiology University of Missouri Columbia Missouri United States of America

Institute of Biology Department of Vertebrate Ecology and Palaeontology Wrocław University of Environmental and Life Sciences Wrocław Poland

Institute of Biostatistics and Analyses Masaryk University Brno Czech Republic

Institute of Immunology Friedrich Loeffler Institute Federal Research Institute for Animal Health Greifswald Insel Riems Germany

Institute of Vertebrate Biology Czech Academy of Sciences Brno Czech Republic

Laboratory of Fungal Genetics and Metabolism Institute of Microbiology Czech Academy of Sciences Prague Czech Republic

Pennsylvania Game Commission Harrisburg Pennsylvania United States of America

United States Department of Agriculture Forest Service Northern Research Station Columbia Missouri United States of America

Zobrazit více v PubMed

Deem SL, Karesh WB, Weisman W. Putting theory into practice: Wildlife health in conservation. Conserv Biol. 2001;15: 1224–1233.

Fey SB, Siepielski AM, Nusslé S, Cervantes-Yoshida K, Hwan JL, et al. Recent shifts in the occurrence, cause, and magnitude of animal mass mortality events. Proc Natl Acad Sci USA. 2015;112: 1083–1088. doi: 10.1073/pnas.1414894112 PubMed DOI PMC

Blehert DS, Hicks AC, Behr M, Meteyer CU, Berlowski-Zier BM, et al. Bat white-nose syndrome: An emerging fungal pathogen? Science. 2009;323: 227 doi: 10.1126/science.1163874 PubMed DOI

Gargas A, Trest MT, Christensen M, Volk TJ, Blehert DS. Geomyces destructans sp. nov. associated with bat white-nose syndrome. Mycotaxon. 2009;108: 147–154.

Lorch JM, Meteyer CU, Behr MJ, Boyles JG, Cryan PM, et al. Experimental infection of bats with Geomyces destructans causes white-nose syndrome. Nature. 2011;480: 376–378. doi: 10.1038/nature10590 PubMed DOI

Minnis AM, Lindner DL. Phylogenetic evaluation of Geomyces and allies reveals no close relatives of Pseudogymnoascus destructans, comb. nov., in bat hibernacula of eastern North America. Fungal Biol. 2013;117: 638–649. doi: 10.1016/j.funbio.2013.07.001 PubMed DOI

Zukal J, Bandouchova H, Bartonicka T, Berkova H, Brack V, et al. White-nose syndrome fungus: A generalist pathogen of hibernating bats. PLoS ONE. 2014;9: e97224 doi: 10.1371/journal.pone.0097224 PubMed DOI PMC

Frick WF, Pollock JF, Hicks AC, Langwig KE, Reynolds DS, et al. An emerging disease causes regional population collapse of a common North American bat species. Science. 2010;329: 679–682. doi: 10.1126/science.1188594 PubMed DOI

Turner GG, Reeder DM, Coleman JTH. A five-year assessment of mortality and geographic spread of white-nose syndrome in North American bats and a look to the future. Bat Research News. 2011;52: 13–27.

Coleman JTH, Reichard JD. Bat white-nose syndrome in 2014: A brief assessment seven years after discovery of a virulent fungal pathogen in North America. Outlooks on Pest Management. 2014;25: 374–377.

Zukal J, Bandouchova H, Brichta J, Cmokova A, Jaron KS, et al. White-nose syndrome without borders: Pseudogymnoascus destructans infection tolerated in Europe and Palearctic Asia but not in North America. Sci Reports. 2016;6: 19829. PubMed PMC

Bandouchova H, Bartonicka T, Berkova H, Brichta J, Cerny J, et al. Pseudogymnoascus destructans: Evidence of virulent skin invasion for bats under natural conditions, Europe. Transbound Emerg Dis. 2015;62: 1–5. doi: 10.1111/tbed.12282 PubMed DOI

Meteyer CU, Buckles EL, Blehert DS, Hicks AC, Green DE, et al. Histopathologic criteria to confirm white-nose syndrome in bats. J Vet Diagn Invest. 2009;21: 411–414. doi: 10.1177/104063870902100401 PubMed DOI

Reeder DM, Frank CL, Turner GG, Meteyer CU, Kurta A, et al. Frequent arousal from hibernation linked to severity of infection and mortality in bats with white-nose syndrome. PLoS ONE. 2012;7: e38920 doi: 10.1371/journal.pone.0038920 PubMed DOI PMC

Warnecke L, Turner JM, Bollinger TK, Lorch JM, Misra V, et al. Inoculation of bats with European Geomyces destructans supports the novel pathogen hypothesis for the origin of white-nose syndrome. Proc Natl Acad Sci USA. 2012;109: 6999–7003. doi: 10.1073/pnas.1200374109 PubMed DOI PMC

Cryan P, Meteyer C, Boyles J, Blehert D. Wing pathology of white-nose syndrome in bats suggests life-threatening disruption of physiology. BMC Biol. 2010;8: 135 doi: 10.1186/1741-7007-8-135 PubMed DOI PMC

Cryan P, Meteyer C, Boyles J, Blehert D. White-nose syndrome in bats: illuminating the darkness. BMC Biol. 2013;11: 47 doi: 10.1186/1741-7007-11-47 PubMed DOI PMC

Cryan PM, Meteyer CU, Blehert DS, Lorch JM, Reeder DM, et al. Electrolyte depletion in white-nose syndrome bats. J Wildlife Dis. 2013;49: 398–402. PubMed

Verant ML, Carol MU, Speakman JR, Cryan PM, Lorch JM, et al. White-nose syndrome initiates a cascade of physiologic disturbances in the hibernating bat host. BMC Physiol. 2014;14: 10 doi: 10.1186/s12899-014-0010-4 PubMed DOI PMC

Warnecke L, Turner JM, Bollinger TK, Misra V, Cryan PM, et al. Pathophysiology of white-nose syndrome in bats: A mechanistic model linking wing damage to mortality. Biol Letters. 2013;9: 20130177. PubMed PMC

Hoyt JR, Sun K, Parise KL, Lu G, Langwig KE, et al. Widespread bat white-nose syndrome fungus, Northeastern China. Emerg Inf Dis. 2016;22: 140. PubMed PMC

Turner GG, Meteyer CU, Barton H, Gumbs JF, Reeder DM, et al. Nonlethal screening of bat-wing skin with the use of ultraviolet fluorescence to detect lesions indicative of white-nose syndrome. J Wildlife Dis. 2014;50: 566–573. PubMed

Flieger M, Bandouchova H, Cerny J, Chudíčková M, Kolarik M, et al. Vitamin B2 as a virulence factor in Pseudogymnoascus destructans skin infection. Sci Reports. 2016;6: 33200. PubMed PMC

Court MH, Robbins AH, Whitford AM, Beck EV, Tseng FS, et al. Pharmacokinetics of terbinafine in little brown myotis (Myotis lucifugus) infected with Pseudogymnoascus destructans. Am J Vet Res. 2017;78: 90–99. doi: 10.2460/ajvr.78.1.90 PubMed DOI

Shuey MM, Drees KP, Lindner DL, Keim P, Foster JT. Highly sensitive quantitative PCR for the detection and differentiation of Pseudogymnoascus destructans and other Pseudogymnoascus species. Appl Environ Microb. 2014;80: 1726–1731. PubMed PMC

Bates D, Maechler M, Bolker B, Walker S. lme4: Linear mixed-effects models using Eigen and S4. R package version 1.1–9. 2015. Available from: https://CRAN.R-project.org/package=lme4.

R Core Team. R: A language and environment for statistical computing. 2016. Available from: http://www.R-project.org/.

Rowlingson B, Diggle P. splancs: Spatial and space-time point pattern analysis. R package version 2.01–38. 2015. Available from: https://CRAN.R-project.org/package=splancs.

Urbanek S. jpeg: Read and write JPEG images. R package version 0.1–8. 2014. Available from: https://CRAN.R-project.org/package=jpeg.

Ives AR, Midford PE, Garland T Jr. Within-species variation and measurement error in phylogenetic comparative methods. Syst Biol. 2007;56: 252–270. doi: 10.1080/10635150701313830 PubMed DOI

Revell LJ. phytools: An R package for phylogenetic comparative biology (and other things). Methods Ecol Evol. 2012;3: 217–223.

Cohen J. A coefficient of agreement for nominal scales. Educ Psychol Meas. 1960;20: 37–46.

McHugh ML. Interrater reliability: the kappa statistic. Biochem Medica. 2012;22: 276–282. PubMed PMC

Fleiss JL. Measuring nominal scale agreement among many raters. Psychol Bull. 1971;76: 378–382.

Gamer M, Lemon J, Singh IFP. irr: Various coefficients of interrater reliability and agreement. R package version 0.84. 2012. Available from: https://CRAN.R-project.org/package=irr.

Martínková N, Bačkor P, Bartonička T, Blažková P, Červený J, et al. Increasing incidence of Geomyces destructans fungus in bats from the Czech Republic and Slovakia. PLoS ONE. 2010;5: e13853 doi: 10.1371/journal.pone.0013853 PubMed DOI PMC

Puechmaille SJ, Wibbelt G, Korn V, Fuller H, Forget F, et al. Pan-European distribution of white-nose syndrome fungus (Geomyces destructans) not associated with mass mortality. PLoS ONE. 2011;6: e19167 doi: 10.1371/journal.pone.0019167 PubMed DOI PMC

Field KA, Johnson JS, Lilley TM, Reeder SM, Rogers EJ, et al. The white-nose syndrome transcriptome: Activation of anti-fungal host responses in wing tissue of hibernating little brown Myotis. PLoS Pathog. 2015;11: e1005168 doi: 10.1371/journal.ppat.1005168 PubMed DOI PMC

Pikula J, Bandouchova H, Novotný L, Meteyer CU, Zukal J, et al. Histopathology confirms white-nose syndrome in bats in Europe. J Wildlife Dis. 2012;48: 207–211. PubMed

Lucan RK, Bandouchova H, Bartonicka T, Pikula J, Zahradnikova A Jr., et al. Ectoparasites may serve as vectors for the white-nose syndrome fungus. Parasit Vectors 2016;9: 16 doi: 10.1186/s13071-016-1302-2 PubMed DOI PMC

Cross SS, Benes K, Stephenson TJ, Harrison RF. Grading in histopathology. Diagnostic Histopathology. 2011;17: 263–267.

McGuire LP, Turner JM, Warnecke L, McGregor G, Bollinger TK, et al. White-nose syndrome disease severity and a comparison of diagnostic methods. EcoHealth. 2016;13: 60–71. doi: 10.1007/s10393-016-1107-y PubMed DOI

Meteyer CU, Barber D, Mandl JN. Pathology in euthermic bats with white nose syndrome suggests a natural manifestation of immune reconstitution inflammatory syndrome. Virulence. 2012;3: 583–588. doi: 10.4161/viru.22330 PubMed DOI PMC

Meteyer CU, Valent M, Kashmer J, Buckles EL, Lorch JM, et al. Recovery of little brown bats (Myotis lucifugus) from natural infection with Geomyces destructans, white-nose syndrome. J Wildl Dis. 2011;47: 618–626. doi: 10.7589/0090-3558-47.3.618 PubMed DOI

Fuller NW, Reichard JD, Nabhan ML, Fellows SR, Pepin LC, et al. Free-ranging little brown myotis (Myotis lucifugus) heal from wing damage associated with white-nose syndrome. Ecohealth. 2011;8: 154–162. doi: 10.1007/s10393-011-0705-y PubMed DOI

Reichard JD, Kunz TH. White-nose syndrome inflicts lasting injuries to the wings of little brown myotis (Myotis lucifugus). Acta Chiropterol. 2009;11: 457–464.

Voigt CC. Bat flight with bad wings: is flight metabolism affected by damaged wings? J Exp Biol. 2013;216: 1516–1521. doi: 10.1242/jeb.079509 PubMed DOI

Lochmiller RL, Deerenberg C. Trade-offs in evolutionary immunology: just what is the cost of immunity? Oikos. 2000;88: 87–98.

Higher antibody titres against Pseudogymnoascus destructans are associated with less white-nose syndrome skin lesions in Palearctic bats

Higher white-nose syndrome fungal isolate yields from UV-guided wing biopsies compared with skin swabs and optimal culture media

Performance of bat-derived macrophages at different temperatures

Active surveillance for antibodies confirms circulation of lyssaviruses in Palearctic bats

Transcriptional host-pathogen responses of Pseudogymnoascus destructans and three species of bats with white-nose syndrome

Comparative eco-physiology revealed extensive enzymatic curtailment, lipases production and strong conidial resilience of the bat pathogenic fungus Pseudogymnoascus destructans

Bat population recoveries give insight into clustering strategies during hibernation

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

Phagocyte activity reflects mammalian homeo- and hetero-thermic physiological states

Resistance is futile: RNA-sequencing reveals differing responses to bat fungal pathogen in Nearctic Myotis lucifugus and Palearctic Myotis myotis

Hibernation temperature-dependent Pseudogymnoascus destructans infection intensity in Palearctic bats

White-nose syndrome detected in bats over an extensive area of Russia

Alterations in the health of hibernating bats under pathogen pressure