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Friday, 3 March 2017

DNA-based detection of the fungal pathogen Geomyces destructans in soils from bat hibernacula

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Year Published

2011

Source

Mycologia. 103(2): 241-246.

Abstract

White-nose syndrome (WNS) is an emerging disease causing unprecedented morbidity and mortality among bats in eastern North America. The disease is characterized by cutaneous infection of hibernating bats by the psychrophilic fungus Geomyces destructans. Detection of G. destructans in environments occupied by bats will be critical for WNS surveillance, management and characterization of the fungal lifecycle. We initiated an rRNA gene region-based molecular survey to characterize the distribution of G. destructans in soil samples collected from bat hibernacula in the eastern United States with an existing PCR test. Although this test did not specifically detect G. destructans in soil samples based on a presence/absence metric, it did favor amplification of DNA from putative Geomyces species. Cloning and sequencing of PCR products amplified from 24 soil samples revealed 74 unique sequence variants representing 12 clades. Clones with exact sequence matches to G. destructans were identified in three of 19 soil samples from hibernacula in states where WNS is known to occur. Geomyces destructans was not identified in an additional five samples collected outside the region where WNS has been documented. This study highlights the diversity of putative Geomyces spp. in soil from bat hibernacula and indicates that further research is needed to better define the taxonomy of this genus and to develop enhanced diagnostic tests for rapid and specific detection of G. destructans in environmental samples.

Keywords

Citation

Lindner, Daniel L.; Gargas, Andrea; Lorch, Jeffrey M.; Banik, Mark T.; Glaeser, Jessie; Kunz, Thomas H.; Blehert, David S. 2011. DNA-based detection of the fungal pathogen Geomyces destructans in soils from bat hibernacula. Mycologia. 103(2): 241-246.

Last updated on: October 20, 2011

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https://www.nrs.fs.fed.us/pubs/39451
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Phylogenetic evaluation of Geomyces and allies reveals no close relatives of Pseudogymnoascus destructans, comb nov, in bat hibernacula of eastern North America

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Year Published

2013

Source

Fungal Biology. 117: 638-649.

Abstract

White-nose syndrome (WNS) of bats, caused by the fungus previously known as Geomyces destructans, has decimated populations of insectivorous bats in eastern North America. Recent work on fungi associated with bat hibernacula uncovered a large number of species of Geomyces and allies, far exceeding the number of described species. Communication about these species has been hindered by the lack of a modern taxonomic evaluation, and a phylogenetic framework of the group is needed to understand the origin of G. destructans and to target closely related species and their genomes for the purposes of understanding mechanisms of pathogenicity. We addressed these issues by generating DNA sequence data for the internal transcribed spacer (ITS) region, nuclear large subunit (LSU) rDNA, MCM7, RPB2, and TEF1 from a diverse array of Geomyces and allies that included isolates recovered from bat hibernacula as well as those that represent important type species. Phylogenetic analyses indicate Geomyces and allies should be classified in the family Pseudeurotiaceae, and the genera Geomyces, Gymnostellatospora, and Pseudogymnoascusshould be recognized as distinct. True Geomyces are restricted to a basal lineage based on phylogenetic placement of the type species, Geomyces auratus. Thus, G. destructans is placed in genus Pseudogymnoascus. The closest relatives of Pseudogymnoascus destructans are members of the Pseudogymnoascus roseus species complex,however, the isolated and long branch of P. destructansindicates that none of the species included in this study are closely related, thus providing further support to the hypothesis that this pathogen is non-native and invasive in eastern North America. Several conidia-producing isolates from bat hibernacula previously identified as members of Pseudeurotium are determined to belong to the genus Leuconeurospora, which is widespread, especially in colder regions. Teberdinia hygrophila is transferred to Pseudeurotium as Pseudeurotium hygrophilum, comb. nov., in accordance with the one name per fungus system of classification, and two additional combinations are made in Pseudogymnoascus including Pseudogymnoascus carnis and Pseudogymnoascus pannorum. Additional sampling from other regions of the world is needed to better understand the evolution and biogeography of this important and diverse group of fungi.

Keywords

Citation

Minnis, Andrew M.; Lindner, Daniel L. 2013. Phylogenetic evaluation of Geomyces and allies reveals no close relatives of Pseudogymnoascus destructans, comb. nov., in bat hibernacula of eastern North America. Fungal Biology. 117: 638-649.

Last updated on: September 13, 2013

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https://www.nrs.fs.fed.us/pubs/44379
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Nightly and yearly bat activity before and after white-nose syndrome on the Fernow Experimental Forest in West Virginia

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Year Published

2013

Publication

Res. Pap. NRS-24. Newtown Square, PA: U.S. Department of Agriculture, Forest Service, Northern Research Station. 17 p.

Abstract

In the central Appalachians, conservation concern about bat communities and their population status has become increasingly more significant with the advent and spread of white-nose syndrome (WNS). However, managers often are hampered in their response to WNS by the lack of information on pre-WNS local distribution, abundance, or activity patterns for most bat species. At the Fernow Experimental Forest (FEF), Tucker County, WV, where bat research has been conducted since the mid-1990s, we acoustically monitored bat activity a total of 20 nights each at four sites for 4 years—3 years before and 1 year after WNS was detected—to better assess those local patterns. Within sampling nights, activity of northern myotis (Myotis septentrionalis) and big brown bats (Eptesicus fuscus) peaked directly after sunset and declined throughout the night, whereas activity of little brown myotis (Myotis lucifugus) and Indiana myotis (Myotis sodalis) had a unimodal distribution that peaked in the middle of the night. Activity of many bat species differed among sample sites and was highest at a small, artificial pond located on a dry ridgetop. Activity of little brown myotis, northern myotis, and Indiana myotis was lower post-WNS than pre-WNS, consistent with the species' precipitous declines previously reported in WNS-affected areas in the Northeast and upper portions of the Mid-Atlantic.

Keywords

Citation

Johnson, Joshua B.; Rodrigue, Jane L.; Ford, W. Mark. 2013. Nightly and yearly bat activity before and after white-nose syndrome on the Fernow Experimental Forest in West Virginia. Res. Pap. NRS-24. Newtown Square, PA: U.S. Department of Agriculture, Forest Service, Northern Research Station. 17 p.

Last updated on: September 30, 2013

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https://www.nrs.fs.fed.us/pubs/44877
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Distribution and environmental persistence of the causative agent of white-nose syndrome, Geomyces destructans, in bat hibernacula of the eastern United States

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  • Lorch, Jeffrey M.
  • Muller, Laura K.
  • Russell, Robin E.
  • O'Connor, Michael
  • Lindner, Daniel L.
  • Blehert, David S.

Year Published

2013

Source

Applied and Environmental Microbiology. 79(4): 1293-1301

Abstract

>White-nose syndrome (WNS) is an emerging disease of hibernating bats caused by the recently described fungus Geomyces destructans. First isolated in 2008, the origins of this fungus in North America and its ability to persist in the environment remain undefined. To investigate the correlation between manifestation of WNS and distribution of G. destructans in the United States, we analyzed sediment samples collected from 55 bat hibernacula (caves and mines) both within and outside the known range of WNS using a newly developed real-time PCR assay. Geomyces destructans was detected in 17 of 21 sites within the known range of WNS at the time when the samples were collected; the fungus was not found in 28 sites beyond the known range of the disease at the time when environmental samples were collected. These data indicate that the distribution of G. destructans is correlated with disease in hibernating bats and support the hypothesis that the fungus is likely an exotic species in North America. Additionally, we examined whether G. destructans persists in infested bat hibernacula when bats are absent. Sediment samples were collected from 14 WNS-positive hibernacula, and the samples were screened for viable fungus by using a culture technique. Viable  G. destructans was cultivated from 7 of the 14 sites sampled during late summer, when bats were no longer in hibernation, suggesting that the fungus can persist in the environment in the absence of bat hosts for long periods of time.

Keywords

Citation

Lorch, Jeffrey M.; Muller, Laura K.; Russell, Robin E.; O’Connor, Michael; Lindner, Daniel L.; Blehert, David S. 2013. Distribution and environmental persistence of the causative agent of white-nose syndrome, Geomyces destructans, in bat hibernacula of the eastern United States. Applied and Environmental Microbiology. 79(4): 1293-1301. doi: 10.1128/​AEM.02939-12

Last updated on: April 18, 2013

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https://www.nrs.fs.fed.us/pubs/43280
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Bat white-nose syndrome: A real-time TaqMan polymerase chain reaction test targeting the intergenic spacer region of Geomyces destructans

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  • Muller, Laura K
  • Lorch, Jeffrey M.
  • Lindner, Daniel L.
  • O'Connor, Michael
  • Gargas, Andrea
  • Blehert, David S.

Year Published

2013

Source

Mycologia. 105(2): 253-259.

Abstract

The fungus Geomyces destructans is the causative agent of white-nose syndrome (WNS), a disease that has killed millions of North American hibernating bats. We describe a real-time TaqMan PCR test that detects DNA from G. destructans by targeting a portion of the multicopy intergenic spacer region of the rRNA gene complex. The test is highly sensitive, consistently detecting as little as 3.3 fg of genomic DNA from G. destructans. The real-time PCR test specifically amplified genomic DNA from G. destructans but did not amplify target sequence from 54 closely related fungal isolates (including 43 Geomyces spp. isolates) associated with bats. The test was further qualified by analyzing DNA extracted from 91 bat wing skin samples, and PCR results matched histopathology findings. These data indicate the real-time TaqMan PCR method described herein is a sensitive, specific, and rapid test to detect DNA from G. destructans and provides a valuable tool for WNS diagnostics and research.

Keywords

Citation

Muller, Laura K; Lorch, Jeffrey M.; Lindner, Daniel L.; O'Connor, Michael; Gargas, Andrea; Blehert, David S. 2013. Bat white-nose syndrome: A real-time TaqMan polymerase chain reaction test targeting the intergenic spacer region of Geomyces destructans. Mycologia. 105(2): 253-259.

Last updated on: April 12, 2013

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https://www.nrs.fs.fed.us/pubs/42974
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A culture-based survey of fungi in soil from bat hibernacula in the eastern United States and its implications for detection of Geomyces destructans, the causal agent of bat white-nose syndrome

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Year Published

2013

Source

Mycologia. 105(2): 237-252.

Abstract

The recent emergence of white-nose syndrome (WNS), a fungal disease causing unprecedented mortality among hibernating bats of eastern North America, has revealed a knowledge gap regarding fungal communities associated with bats and their hibernacula. We used culture-based techniques to investigate the diversity of fungi in soil samples collected from 24 bat hibernacula in the eastern United States. Ribosomal RNA regions (internal transcribed spacer and partial intergenic spacer) were sequenced to preliminarily characterize isolates. Geomyces species were one of the most abundant and diverse groups cultured, representing approximately 33% of all isolates. Geomyces destructans was isolated from soil samples from three hibernacula in states where WNS is known to occur, and many of the other cultured Geomyces isolates likely represent undescribed taxa. Further characterization of the diversity of fungi that occur in hibernacula will both facilitate an improved understanding of the ecology of G. destructans within this complex fungal community and provide an opportunity to identify characteristics that differentiate G. destructans from non-pathogenic relatives.

Keywords

Citation

Lorch, Jeffrey M.; Lindner, Daniel L.; Gargas, Andrea; Muller, Laura K; Minnis, Andrew M.; Blehert, David S. 2013. A culture-based survey of fungi in soil from bat hibernacula in the eastern United States and its implications for detection of Geomyces destructans, the causal agent of bat white-nose syndrome. Mycologia. 105(2): 237-252.

Last updated on: April 12, 2013

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https://www.nrs.fs.fed.us/pubs/42973
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Molecular characterization of a heterothallic mating system in Pseudogymnoascus destructans, the fungus causing white-nose syndrome of bats

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Year Published

2014

Source

Genes|Genomes|Genetics. 4(9): 1755-1763.

Abstract

White-nose syndrome (WNS) of bats has devastated bat populations in eastern North America since its discovery in 2006. WNS, caused by the fungus Pseudogymnoascus destructans, has spread quickly in North America and has become one of the most severe wildlife epidemics of our time. While P. destructans is spreading rapidly in North America, nothing is known about the sexual capacity of this fungus. To gain insight into the genes involved in sexual reproduction, we characterized the mating-type locus (MAT) of two Pseudogymnoascus spp. that are closely related to P. destructans and homothallic (self-fertile). As with other homothallic Ascomycota, the MAT locus of these two species encodes a conserved a-box protein (MAT1-1-1) as well as two high-mobility group (HMG) box proteins (MAT1-1-3 and MAT1-2-1). Comparisons with the MATlocus of the North American isolate of P. destructans (the ex-type isolate) revealed that this isolate of P. destructans was missing a clear homolog of the conserved HMG box protein (MAT1-2-1). These data prompted the discovery and molecular characterization of a heterothallic mating system in isolates of P. destructans from the Czech Republic. Both mating types of P. destructans were found to coexist within hibernacula, suggesting the presence of mating populations in Europe. Although populations of P. destructansin North America are thought to be clonal and of one mating type, the potential for sexual recombination indicates that continued vigilance is needed regarding introductions of additional isolates of this pathogen.

Keywords

Citation

Palmer, Jonathan M.; Kubatova, Alena; Novakova, Alena.; Minnis, Andrew M.; Kolarik, Miroslav; Lindner, Daniel L. 2014. Molecular characterization of a heterothallic mating system in Pseudogymnoascus destructans, the fungus causing white-nose syndrome of bats. Genes|Genomes|Genetics. 4(9): 1755-1763.

Last updated on: September 26, 2014

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https://www.nrs.fs.fed.us/pubs/46717
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Bat Activity in Harvested and Intact Forest Stands in the Allegheny Mountains

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  • Owen, Sheldon F.
  • Menzel, Michael A.
  • Edwards, John W.
  • Ford, W. Mark
  • Menzel, Jennifer M.
  • Chapman, Brian R.
  • Wood, Petra Bohall
  • Miller, Karl V.

Year Published

2004

Publication

Northern Journal of Applied Forestry. 21(3): 154-159.

Abstract

Keywords

Citation

Owen, Sheldon F.; Menzel, Michael A.; Edwards, John W.; Ford, W. Mark; Menzel, Jennifer M.; Chapman, Brian R.; Wood, Petra Bohall; Miller, Karl V. 2004. Bat Activity in Harvested and Intact Forest Stands in the Allegheny Mountains. Northern Journal of Applied Forestry. 21(3): 154-159.

Last updated on: August 22, 2006

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https://www.nrs.fs.fed.us/pubs/6921
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Relating bat species presence to simple habitat measures in a central Appalachian forest

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Year Published

2005

Publication

Biological Conservation. 126: 528-539.

Abstract

We actively sampled the bat community at 63 sites using detection and non- detection metrics on the Fernow Experimental Forest (FEF) in the central Appalachians of West Virginia using Anabat acoustical equipment May-June 2001-2003 to relate species presence to simple habitat measures such as proximity to riparian areas, forest canopy cover, forest canopy gap width, and forest canopy height. We acoustically detected eight species on the FEF, including the endangered Myotis sodalis. The presence of Lasiurus cinereus, M. lucifugus, M. sodalis, and Pipistrellus subflaus was associated more with riparian areas than upland areas. Both univariate comparisons and multiple logistic regression modeling showed that the probability that clutter-adapted foraging species such as M. septentrionalis and M. sodalis would be detected was greater as forest canopy cover increased or forest canopy gap size decreased, whereas the opposite was true for open-adapted foraging species such as Eptesicus fuscus and L. cinereus The overall proportion of unidentifiable bat echolocation sequences to those identified to species was related to upland sites with increasing forest canopy cover indicating some sampling bias between cluttered and uncluttered habitats. However, given sufficient sample points, bat community surveys using acoustical detection show the ability to quickly develop generalized habitat associations for rugged areas such as the central Appalachians where traditional mist-net survey efforts often are logistically difficult and are lacking in scope. Moreover, these acoustical surveys also could lend themselves to species-specific predictive mapping of foraging habitat as well as allowing researchers to formulate testable hypotheses about detailed bat habitat relationships to be definitively tested with radio-telemetry techniques.

Keywords

Citation

Ford, W. Mark; Menzel, Michael A.; Rodrigue, Jane L.; Menzel, Jennifer M.; Johnson, Joshua B. 2005. Relating bat species presence to simple habitat measures in a central Appalachian forest. Biological Conservation. 126: 528-539.

Last updated on: October 18, 2007

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https://www.nrs.fs.fed.us/pubs/7746
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Bat Activity in a Forest Landscape of central Massachusetts

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Year Published

2005

Source

Northeastern Naturalist 12(4):447-462

Abstract

Nine species of bat are known to occur across the six New England a states, but most aspects of their natural history, such as foraging habitat use, are poorly understood. Recent published research has documented the importance of still-water habitats as foci of bat flight activity. To better understand and document habitat use in southern New England, we used the AnaBat II acoustical monitoring system to assess species composition and relative levels of summer flight activity. Active acoustic surveys were conducted in six habitat types on the Quabbin Reservation in central Massachusetts in 2003 and 2004. Bat flight activity, as measured by numbers of echolocation call sequences, was high, with an average of 24 search-phase and 4 feeding-buzz calls per 20-minute survey period. Myotis lucifugus (little brown bat) was the most commonly recorded species. Bat flight activity was high over all still-water habitats, but greatest over large ponds. Large-bodied bats, such as Eptesicus fuscus (big brown bat), were recorded more often in open, structurally uncluttered habitats. Of the small-bodied bats, little brown bats were ubiquitous, whereas Myotis septentrionalis (northern myotis) was most common in structurally cluttered habitats of seasonal forest (vernal) pools and along forest streams. Generalized habitat associations among the bat species we recorded are similar to those reported for other New England forest sites. The Quabbin Reservation is an excellent site to continue examining bat-habitat relationships because of the abundance and diversity of aquatic habitats, in both cluttered closed-canopy and uncluttered open-canopy settings.

Citation

Brooks, Robert T.; Ford, W. Mark 2005. Bat Activity in a Forest Landscape of central Massachusetts. Northeastern Naturalist 12(4):447-462
Last updated on: June 14, 2006

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https://www.nrs.fs.fed.us/pubs/3801
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