Mycobiome of the Bat White Nose Syndrome (WNS) Affected Caves and Mines reveals High Diversity of Fungi and Local Adaptation by the Fungal Pathogen Pseudogymnoascus (Geomyces) destructans

The investigations of the bat White Nose Syndrome (WNS) have yet to provide answers as to how the causative fungus Pseudogymnoascus (Geomyces) destructans (Pd) first appeared in the Northeast and how a single clone has spread rapidly in the US and Canada. We aimed to catalogue Pd and all other fungi (mycobiome) by the culture-dependent (CD) and culture-independent (CI) methods in four Mines and two Caves from the epicenter of WNS zoonotic. Six hundred sixty-five fungal isolates were obtained by CD method including the live recovery of Pd. Seven hundred three nucleotide sequences that met the definition of operational taxonomic units (OTUs) were recovered by CI methods. Most OTUs belonged to unidentified clones deposited in the databases as environmental nucleic acid sequences (ENAS). The core mycobiome of WNS affected sites comprised of 46 species of fungi from 31 genera recovered in culture, and 17 fungal genera and 31 ENAS identified from clone libraries. Fungi such as Arthroderma spp., Geomyces spp., Kernia spp., Mortierella spp., Penicillium spp., and Verticillium spp. were predominant in culture while Ganoderma spp., Geomyces spp., Mortierella spp., Penicillium spp. and Trichosporon spp. were abundant is clone libraries. Alpha diversity analyses from CI data revealed that fungal community structure was highly diverse. However, the true species diversity remains undetermined due to under sampling. The frequent recovery of Pd indicated that the pathogen has adapted to WNS-afflicted habitats. Further, this study supports the hypothesis that Pd is an introduced species. These findings underscore the need for integrated WNS control measures that target both bats and the fungal pathogen.Comment: 59 pages, 7figure

Clonal Genotype of Geomyces destructans among Bats with White Nose Syndrome, New York, USA

The dispersal mechanism of Geomyces destructans, which causes geomycosis (white nose syndrome) in hibernating bats, remains unknown. Multiple gene genealogic analyses were conducted on 16 fungal isolates from diverse sites in New York State during 2008–2010. The results are consistent with the clonal dispersal of a single G. destructans genotype

VecTest as Diagnostic and Surveillance Tool for West Nile Virus in Dead Birds

The VecTest WNV assay is adequate for diagnostic and surveillance purposes in American Crows, Blue Jays, and House Sparrows

Bat White-Nose Syndrome: An Emerging Fungal Pathogen?

White-nose syndrome (WNS) is a condition associated with an unprecedented bat mortality event in the northeastern United States. Since the winter of 2006*2007, bat declines exceeding 75% have been observed at surveyed hibernacula. Affected bats often present with visually striking white fungal growth on their muzzles, ears, and/or wing membranes. Direct microscopy and culture analyses demonstrated that the skin of WNS-affected bats is colonized by a psychro-philic fungus that is phylogenetically related to Geomyces spp. but with a conidial morphology distinct from characterized members of this genus. This report characterizes the cutaneous fungal infection associated with WNS

Phylogenetic relationships among ITS phylotypes for OTUs recovered by culture-independent investigation.

<p>Phylotypes recovered during this study are shown in bold type. Sequence code prefix denotes location. ACV, Aeolus Cave, Bennington County, VT; BME, Barton Mine, Essex County, NY; GMW, Graphite Mine, Warren County, NY; HME, Hitchcock Mine, Essex County, NY; WMU, Williams Preserve Mine, Ulster County, NY; HCA, Hailes Cave, Albany. Blue font denotes the recovered phylotypes; red font denotes the <i>Geomyces</i> spp. and <i>Pseudogymnoascus</i> spp., and black font denotes the ITS sequences of reference strains retrieved from GenBank database. Three topologies are supported by the program Mega 5.1 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108714#pone.0108714-Tamura1" target="_blank">[44]</a>. The numbers at node indicate the bootstrap percentages of 1,000 resamples.</p

Phyla distribution in culture-independent (CI) clones.

<p>A) Relative proportions of different phyla of all clones; B) Relative proportions of different phyla of LSU clones; C) Relative proportions of different phyla of ITS clones. The zero percent assigned in pie chart represents clones with less than 1% relative distribution.</p

Overlap in fungal community composition across bat WNS-afflicted environmental samples.

<p>Venn diagram demonstrates the degree of overlap of OTU similarity among niches. OTUs were defined using 97% genetic similarity cut-off. A) Venn diagram displays the overlapping OTUs among ACV, BME, WMU, and HCA; B) Venn diagram displays the overlapping OTUs among GMW, BME, WMU, and HME. ACV, Aeolus Cave, Bennington County, VT; BME, Barton Mine, Essex County, NY; GMW, Graphite Mine, Warren County, NY; HME, Hitchcock Mine, Essex County, NY; WMU, Williams Preserve Mine, Ulster County, NY; HCA, Hailes Cave, Albany.</p