ArticlePDF Available

White-nose syndrome decontamination procedures for backcountry subterranean projects

Abstract and Figures

White-nose syndrome (WNS), a disease caused by the fungal pathogen Pseudogymnoascus destructans, is responsible for the population decline of at least 10 subterranean hibernating bat species in eastern North America and has recently been confirmed in the northwestern United States. The US Fish and Wildlife Service (USFWS), in concert with other federal and state agencies and university personnel, has developed, and periodically updates, a WNS Decontamination Protocol (e.g., USFWS 2016) for working in the subterranean realm. The protocol is a combination of scientifically tested and untested steps that provide a foundational framework for protecting hibernating bats from inadvertent human-assisted transmission of WNS to uninfected hibernacula. However, it does not specifically address extended backcountry research needs. During four research trips to Grand Canyon–Parashant National Monument, Arizona, from 2011 through 2012, colleagues and I tested and refined backcountry WNS decontamination procedures. The procedures presented here are developed to complement the WNS Decontamination Protocol; provide a stepwise method for disinfecting equipment, clothing, and personnel; and proactively address WNS containment concerns in the backcountry.
Content may be subject to copyright.
in the mortalities of more than five million bats (US-
FWS 2012a) in 33 states and five Canadian provinces
(WDFW 2016). Bat species presently affected by this epizootic
include the cave myotis (Myotis velifer), Townsend’s big-eared bat
(Corynorhinus townsendii), tricolored bat (Perimyotis subflavus),
big brown bat (Eptesicus fuscus), little brown bat (Myotis luci-
fugus), eastern small-footed bat (Myotis leibii), the federally listed
(by the US Fish and Wildlife Service; USFWS) as threatened
northern long-eared bat (Myotis septentrionalis), the federally
listed as endangered Indiana bat (Myotis sodalis), the federally
listed as endangered gray bat (Myotis grisescens) (Meteyer et al.
2009; Turner et al. 2011; USFWS 2014; USFWS 2015b), and the
federally listed as threatened northern long-eared bat (Myotis
septentri onalis; USFWS 2015b). In Canada, the Canadian Coop-
erative Wildlife Health Centre emergency-listed the tricolored
(Perimyotis subflavus), little brown (Myotis lucifugus), and north-
ern long-eared (Myotis septentrionalis) bats as endangered due to
population declines associated with WNS (CCWHC 2014).
White-nose syndrome is caused by the cold-adapted fungus
Pseudogymnoascus destructans (Minnis et al. 2013). When
P. destructans is fully expressed, it presents as a white fungus that
attacks the epithelial layer and digests live skin cells of the rostral
muzzle (furless area around the nose), ears, wing membrane,
forearms, and uropatagium (tail membrane between the thighs) of
hibernating bats (Meteyer et al. 2009; Blehert et al. 2009; Gargas
et al. 2009; Cryan et al. 2010; Foley et al. 2011; fig. 1C and D). Be-
cause numerous dematophytes (pathogenic fungi) occur on bats,
histology is required to confirm the presence of WNS (Meteyer et
al. 2009). However, long-wave ultraviolet (UV) light (wavelength
366–385 nm) may now be used to detect WNS on hibernating bats
in the field. Bats with expressed effects of P. destructans present
with a distinct orange-yellow fluorescence in the affected areas
under UV light (Turner et al. 2014).
Spread of the pathogen
Since it was first documented in Howe Cave, New York, in 2007,
WNS has spread from upstate New York northwestward through
southern Ontario, Canada, northeastward to Nova Scotia, south-
ward to Missouri and Arkansas, and westward through northern
Texas (USFWS 2015a; TPWD 2017). Last year WNS was detected
in King County, Washington, resulting in a 1,300-mile (2,092
km) leap from its previous westernmost locality (WDFW 2016).
Figure 2 shows the current extent, including 2017 range expan-
sion into Minnesota, Nebraska, and Texas. Updated maps of the
spread are maintained at
The primary vector believed responsible for the westward ex-
pansion of WNS is bat-to-bat transmission (e.g., Frick et al. 2010;
Lorch et al. 2011; Puechmaille et al. 2011; Turner et al. 2011). Turner
et al. (2011) suggest transmission likely occurs during fall swarm-
ing and interhibernacula movements of infected bats. Therefore,
to manage for and develop mitigation strategies against WNS on a
landscape scale, we will need to understand movements between
fall swarming and winter hibernacula roosts as well as roost
switching during the hibernation period.
Evidence suggests white-nose syndrome (and the causative agent, P.
destructans) was introduced from Eurasia to North America by hu-
mans. WNS has been identified in 13 bat species from cave hibernac-
State of the Science
White-nose syndrome decontamination procedures for
backcountry subterranean projects
By J. Judson Wynne
White-nose syndrome (WNS), a disease caused by the fungal
Pseudogymnoascus destructans
, is responsible for the
population decline of at least six subterranean hibernating bat
species in eastern North America and has recently been confirmed
in the northwestern United States. The US Fish and Wildlife Service
(USFWS), in concert with other federal and state agencies and
university personnel, has developed, and periodically updates, a
WNS Decontamination Protocol (e.g., USFWS 2016) for working
in the subterranean realm. The protocol is a combination of
scientifically tested and untested steps that provide a foundational
framework for protecting hibernating bats from inadvertent human-
assisted transmission of WNS to uninfected hibernacula. However, it
does not specifically address extended backcountry research needs.
During four research trips to Grand Canyon–Parashant National
Monument, Arizona, from 2011 through 2012, colleagues and I
tested and refined backcountry WNS decontamination procedures.
The procedures presented here are developed to complement the
WNS Decontamination Protocol; provide a stepwise method for
disinfecting equipment, clothing, and personnel; and proactively
address WNS containment concerns in the backcountry.
Key words
bats, caves, mines,
Pseudogymnoascus destructans
0 100 200 400 Miles
White-nose Syndrome Spread
Confirmed = Black outlines
Suspected = Blue outlines
Figure 1. (A and B) Two clusters of healthy hibernating Townsend’s big-eared bats at Grand Canyon– Parashant National Monument, Arizona.
Examples of (C) a tricolored bat in a cave at Cloudland State Park, Georgia, 2013, and (D) a little brown bat at the Greeley Mine, Vermont,
2009, with fully expressed white-nose syndrome.
Figure 2. Known distribution of white-nose syndrome in the United States and Canada by county and district as of March 2017. On 31 March 2016,
WNS was confirmed in King County, Washington, resulting in a 1,300-mile (2,092 km) leap from its previous westernmost locality (map inset, top
left). Locations of counties in Texas and Minnesota are approximate. (Counties/municipalities are listed by name, year, and state/province in a table
available online.)
ula in several European countries (Wibbelt et al. 2010; Puechmaille
et al. 2011; Zukal et al. 2014) and 6 bat species in eastern China (Hoyt
et al. 2016) with no reported mass mortalities. Humans were the
most likely vector for the introduction of P. destructans from Europe
(Frick et al. 2010; Blehert et al. 2011; Foley et al. 2011) or temperate
regions of Asia to the northeastern United States. The New York
Department of Environmental Conservation, Wildlife Pathology
Unit, detected a fungal conidia (asexually reproducing spore) with a
morphology similar to P. destructans on caving gear tested immedi-
ately after exiting a WNS confirmed site (Okoniewski et al. 2010).
In addition to the initial introduction of P. destructans to North
America, humans likely contribute to the dispersal of this epizoot-
ic pathogen. Early on, Wolf and Wolf (1947) identified humans as a
vector for pathogenic fungi. On Hawaiʻi, Baker (1966) identified at
least 65 different species of fungi from the shoes of travelers (both
being worn and within luggage) arriving from outside debarkation
points. Of these, 15 species were unknown to Hawaiʻi. The most
recent range expansion of WNS to Washington State, involving a
1,300-mile (2,092 km) distance between the closest known WNS
affected area and the detection site, cannot be explained by natural
bat movements. It probably represents a human-assisted range
expansion event. WNS was likely introduced to Washington on
contaminated clothing or caving equipment originating from east-
ern North America, Europe, or northern Asia.
Disease containment
Given that direct management of bat-to-bat transmission is not
possible, scientists and land managers have focused on devel-
oping and implementing procedures to reduce the potential for
human-caused dispersal of this pathogen to uninfected areas.
Since winter 2008, a multiagency team led by the USFWS has pro-
vided a protocol for WNS decontamination (e.g., USFWS 2016)
for regions where the disease is confirmed, suspected, or uncon-
firmed. This protocol provides guidelines for laundering clothing
for 10 minutes or immersing in 131° F (55° C) water for 20 minutes,
and recommends decontamination of other washable gear and
equipment following the manufacturer’s cleaning guidelines. It
also requires all equipment be used in a site-specific manner (e.g.,
no equipment from the WNS confirmed or suspected area may be
used in an unaffected area; USFWS 2016).
In the WNS affected areas (presently eastern North America
and one locality in the Pacific Northwest), either underground
research on most state and federal lands has been restricted or
compliance with the current WNS Decontamination Protocol is
required. For example, Indiana bat winter survey protocols limit
researchers to inventorying hibernacula every other year (Hicks
et al. 2009). On US Forest Service lands in Arkansas, a five-year
moratorium was recently passed on the three national forests to
protect bat populations (USDAFS 2014). In general, the National
Park Service (NPS) requested that cave resource management
plans for all park units include provisions to reduce the threat
of human-assisted transmission of WNS; these provisions may
involve closure of some caves. Where the risk of spreading
P. destructans into or out of parks by visitors can be minimized
(e.g., through screening, decontamination, and the permitting
process), most NPS-managed caves remain open (NPS 2010).
While the most recent WNS Decontamination Protocol (USFWS
2016) has explicit language regarding decontamination procedures,
implementation remains at the discretion of the regulatory and
resource management agencies under which land management
jurisdiction resides. These entities may choose to develop addenda
and supplemental documentation to accompany the most recent
WNS Decontamination Protocol. Thus, regulatory and resource
managers have the flexibility to incorporate additional require-
ments or exemptions based upon the perceived threat level of
WNS in a given region, local conditions, logistical constraints with
implementation, and the best available scientific information.
Need for backcountry decontamination methods
In backcountry settings, cave researchers and resource managers
must plan for a variety of environmental concerns associated
with proper disposal of WNS decontaminated water-chemical
mixtures, as well as logistical constraints on both chemical and
water use and transportation. Dumping chemical products, such
as quaternary ammonium compounds, may have negative envi-
ronmental impacts. These activities are often illegal on state and
federally administered lands in the United States (e.g., NPS 2006).
Preparing solutions for gear submersion requires a significant
amount of water, and packing large amounts of water is often
difficult to impossible in remote backcountry settings. The de-
contamination protocol (USFWS 2016) is typically implemented
upon return from the field—in most cases, on a daily basis. Many
backcountry trips are up to two weeks in duration, and it is not
possible to wash clothing daily. Moreover, it is difficult to sub-
merse equipment in water-chemical mixtures on a regular basis
while in the field. Doing so is logistically challenging when a large
number of sites are visited during a specific research trip, a large
number of field personnel are participating in the field, and when
field personnel pack all equipment into and out of remote areas.
Procedure development and refinement
Using the earlier 2011 (USFWS 2011a, b, c) and later the 2012 de-
contamination protocols (USFWS 2012b), NPS resource manag-
ers, research technicians, and I applied these techniques to the
backcountry to devise methods for effectively decontaminating
gear in areas where logistics were challenging and resources limit-
ed. Ten different field personnel tested and refined these tech-
niques during four research trips (February, June, and September
2011 and February 2012) at Grand Canyon–Parashant National
Monument, northwestern Arizona—an area where WNS does not
occur. We applied incrementally improved versions of these pro-
cedures during the four different research trips, which totaled 100
individual applications in the field (table 1). Discussions were held
at the end of each field day and during a post-expedition debrief-
ing whereby problem areas with applying these procedures were
captured and improvements were made accordingly. Additionally,
we compared DuPont™ brand Tyvek® and ProShield® model dis-
posable coveralls specifically for durability in constricted passage-
ways over long hours of use underground.
Table 1. Number of research trips with related information
for testing and refining backcountry white-nose syndrome
decontamination procedures
Trip Date
Ma r c h 20 11 311 33
June 2011 7 5 35
September 2011 5 4 20
Ma rc h 20 12 4 3 12
Total 12123 10 0
Note: The p rocedure s were teste d and develo ped at Grand Ca nyon– Parashant N ational
Monument, Arizona.
1 Several of t he same team m embers p articip ated on mult iple trips; t herefore, th e total
number prov ided is for t he number of in dividual s who part icipated in t his work.
2 The numbe r of times eac h team appli ed the proce dures.
3 The total nu mber of tim es procedu res were app lied per tri p, calculate d by the numbe r
of perso nnel times t he number of p rocedures u sed in the fi eld.
Through rigorous field testing, we developed a set of stepwise
procedures for disinfecting field equipment and provide recom-
mendations for washing and cleaning exposed parts of the body, as
well as disinfecting and storing gear after daily field operations. We
present this information as four appendixes combining checklists
and protocols in a format that can easily be printed and laminated
for field use. Appendix I lists required supplies, equipment, and
explanations. Appendix II recommends fieldwork preparations.
Appendix III describes procedures prior to entry and after exiting
a study site (i.e., cave or mine), while Appendix IV provides proce-
dures for full decontamination (i.e., prior to moving from one study
site to another). The appendixes follow on pages 57–62.
To prevent the potential for contamination of clean gear that would
be used to facilitate our return to the vehicles and camp (e.g., hiking
boots, backpacks, and satellite phones), we employed a three-con-
tainment-zone approach (Appendix II, Section 3). The three con-
tainment areas are the (1) clean zone, an area to stage non-cave-re-
lated gear (e.g., backpacks, extra water bottles, satellite phone, and
other equipment), and to change into clean coveralls, boots, and
other equipment once the person has left the intermediate zone;
(2) decontamination zone, the location for staging disinfecting
equipment and supplies, and using them to clean exposed parts of
the body, stripping off and isolating coveralls, and changing into
clean clothing; and (3) intermediate zone, the area for staging
clean boots and a clean change of clothes (for the hike back to vehi-
cles/camp) isolated in a ziplock bag, as well as cleaned gear that can
be moved into the clean zone once decontamination procedures
are completed. When used correctly, this approach should enable
workers to stage and isolate contaminated gear and maintain clean
equipment in different areas at a safe distance apart.
When the performance of Tyvek® and ProShield® coveralls was
compared, we found both suit types sustained breaches by abrad-
ing and tearing when navigating constricted passageways. Although
breaches in suits were repaired as detected using duct tape, this
resulted in the introduction of pieces of coverall fabric into the cave
environment. Thus, the use of both suits resulted in physical “litter”
and a chemical contamination concern for the subterranean envi-
ronment. During all field trials, team members attempted to collect
and remove all coverall debris as encountered.
We also encountered problems when using inexpensive duct tape.
Short-term placement (<5 minutes) in direct sunlight on 81°F
(27°C) clear days resulted in the adhesive melting and the tape be-
coming useless until it cooled. We did not experience any problems
with short-term placement of Gorilla® duct tape in direct sunlight.
The 2012 WNS Decontamination Protocol suggests covering
electronic equipment with plastic wrap such as clear plastic bags
(USFWS 2012b). We attempted to cover our digital single-lens
camera in plastic wrap; however, the plastic wrap made it difficult
to use the buttons and view the LCD display. Additionally, without
the use of duct tape (which further restricts one’s ability to use the
camera), the plastic wrap does not adhere to the camera. Though
it was not tested, clear packaging tape used with plastic wrap may
help. For photographing hibernating bats during our February
2012 trip, we chose to use the camera without any barrier, wiping
it down with isopropyl alcohol (70%) wipes after use and placing
it out of the camera box so that it was completely dry before being
stored. The 2016 WNS Decontamination Protocol recommends
site-specific use for this type of equipment (USFWS 2016). Given
that WNS has not been identified in northwestern Arizona, our
approach was compliant with the new recommendations.
The backcountry techniques proposed here were developed to
complement the most recent WNS Decontamination Protocol
(USFWS 2016). This addendum provides stepwise procedures
and eliminates much of the guesswork for first-time users decon-
taminating clothing and equipment. Although they were devel-
oped in response to backcountry subterranean research needs in
the southwestern United States, these methods are applicable for
all backcountry research projects.
These procedures are dynamic, and should be reviewed and
modified as disinfectants and disinfection techniques are im-
proved, or when additional information prompts further revision.
One method for improving these techniques may be through
working with professionals outside the disciplines of microbiolo-
gy and wildlife science such as hazardous materials professionals
and military personnel. Both have long histories of dealing with
biological threats and developing techniques to isolate pathogens
from human populations. Through such a collaboration, we may
be able to further advance our ability to more effectively decon-
taminate equipment and personnel and thus better protect bat
To reduce the likelihood of human-to-hibernacula transmission
of WNS, caves should not be entered unless either a research
question or administrative issue warrants such entry. If so, we
recommend adhering to the most recent WNS Decontamination
Protocol (e.g., USFWS 2016) and following the guidelines, adden-
da, and other supplemental documentation issued by state and
federal regulatory and resource management agencies that have
jurisdiction over the lands where the work will take place.
While the backcountry procedures presented in the four appen-
dixes provide a stepwise approach for decontaminating equip-
ment and personnel (in compliance with the WNS Decontam-
ination Protocol), there are limitations. For many cave research
projects, workers must use expensive, often irreplaceable elec-
tronic equipment (e.g., meteorological instruments, laser distance
finders, and hammer drills). We recommend users of this type
of equipment explore methods to best create a buffer between
the equipment and the cave environment. The WNS Decontam-
ination Protocol suggests site-specific dedication of equipment
(USFWS 2016). Though expensive, this certainly eliminates the
need to apply decontamination protocols for most gear and thus
may be the best approach.
When used in constricted passageways, both brands of coveralls
(Tyvek® and ProShield®) that we tested were subject to breach-
es, tears, and the resultant introduction of fabric into the cave
environment. Thus, both suit types are of limited use within caves
requiring belly crawling or walking through constricted passage-
ways. We should further acknowledge that neither suit type is
designed for the rigors of the cave environment.
In caves with constricted passageways, we do not recommend
using either type of disposable coverall. Instead, we recommend
the use of reusable ballistic nylon coveralls (which are designed
for use in caves), following USFWS (2016) site-specific desig-
nation procedures. However, in backcountry settings it may be
challenging to portage multiple pairs of nylon coveralls, and this
approach would require the same decontamination procedures
applied to other caving equipment when moving between study
sites (USFWS 2016).
Regarding vertical climbing equipment, experiments have been
conducted to test the strength of only Sterling® climbing ropes
and one-inch tubular webbing; Barton (2009) was able to demon-
strate that after numerous WNS decontamination treatments, the
strength of this equipment was not affected. There are more than
a dozen manufacturers that make rope and perhaps twice as many
companies that manufacture climbing harnesses, webbing, and
other such equipment. Conducting experiments similar to Barton
(2009) on all ropes, webbing, harnesses, and other gear made by
different companies has not been attempted. General care and
cleaning of ropes (e.g., Cox and Fulsaas 2003) and harnesses (e.g.,
Black Diamond Journal 2010) involves machine washing on the
gentle cycle or hand washing in a bathtub using mild soap with no
harsh chemicals.
The US Fish and Wildlife Service (2016) recommends either that
rope and webbing be dedicated to a single cave or the cave should
not be entered; ropes and harnesses should be cleaned follow-
ing the manufacturer’s specifications after use at each study site.
We suggest using ropes nearing retirement or those designated
for site-specific applications (USFWS 2016); subsequently, these
ropes may be retired after use or used site-specifically. In areas
where WNS is neither confirmed nor suspected, it may be possi-
ble to use ropes, webbing, harnesses, and other vertical gear and
rope rigging equipment with soft components at different sites af-
ter cleaning this equipment following the manufacturer’s recom-
mendations. However, the manner in which vertical equipment
is used and the frequency of cleaning will be at the discretion of
the jurisdictional regulatory or resource management agency (and
according to the manufacturer’s recommendations).
Management implications
The backcountry WNS decontamination procedures described
here follow the current decontamination protocol (USFWS
2016), as well as previous versions of the protocol (USFWS 2011a,
b, c; 2012b). The approach presented here is the first to outline
a stepwise procedure for implementing WNS decontamination
strategies in the backcountry. Although these procedures were
developed for areas outside the WNS confirmed and suspected
areas (i.e., the western United States), they are applicable in con-
firmed or suspected areas as well.
As time progresses and we learn more about the natural history
characteristics and habitat requirements of P. destructans, we will
be able to use this information to further improve decontami-
nation procedures. Additionally, as more information becomes
available regarding the fall and winter movements of bat species
that hibernate in caves and mines, we will continue to improve
our abilities to manage bats and their roost sites under a WNS
The author extends much gratitude to Jeff Bradybaugh (formally
of Grand Canyon–Parashant National Monument) for support-
ing this research, and Rosie Pepito and Eathan McIntyre, also at
the national monument, for permitting the author to explore and
develop these procedures. Field personnel involved in proce-
dure testing and refinement include Zach Fitzner, Greg Flores,
Nicholas Glover, Todd Heckman, Pete Kelsey, Mike Kotanian,
Bill Mason, Eathan McIntyre, Timothy Titus, Abigail Tobin, and
Shawn Thomas. Hazel Barton (University of Akron), Jeremy
Coleman and Richard Geboy (USFWS), Kevin Drees and Jeff
Foster (Department of Molecular, Cellular, and Biomedical
Sciences; University of New Hampshire–Durham), Jim Kennedy
(Kennedy Above/Under Ground LLC), Joe Merritt (University
of Illinois), Pat Ormsbee (USDA Forest Service), Abigail Tobin
(Northern Arizona University), Shawn Thomas (Bat Conserva-
tion International), and Paul Whitefield (National Park Service)
provided valuable comments and recommendations leading to
the improvement of these procedures. Two anonymous reviewers
provided additional comments to further strengthen this article.
This work was supported through a cooperative agreement be-
tween the National Park Service and Northern Arizona University
(CP-CESU Projects R8230100244 and R8230110012).
While these procedures were improved through communication
with employees of several state and federal agencies, this article
does not necessarily reflect the positions or viewpoints of any of
these state governments or of the United States government on
this important issue.
Literature cited
Baker, G. E. 1966. Inadvertent distribution of fungi. Canadian Journal of
Microbiology 12:109–112.
Barton, H. A. 2009. Safety of Sterling rope following white nose syndrome
decontamination protocol. Unpublished report submitted to National
Speleological Society.
Black Diamond Journal. 2010. QC Lab: Strength of a worn belay loop and
when to retire a harness.
Blehert, D. S., A. C. Hicks, M. Behr, C. U. Meteyer, B. Berlowski-Zier,
E. L. Buckles, J. T. H. Coleman, S. R. Darling, A. Gargas, R. Niver,
J. C. Okoniewski, R. J. Rudd, and W. B. Stone. 2009. Bat white-nose
syndrome: An emerging fungal pathogen? Science 323:227.
Blehert, D. S., J. M. Lorch, A. E. Ballmann, P. M. Cryan, and C. U. Meteyer.
2011. Bat white-nose syndrome in North America. Microbe 6:267–273.
CCWHC (Canadian Cooperative Wildlife Health Centre). 2014.
Little brown myotis, northern myotis and tri-colored bat.
CCWHC, Minister of Environment Canada. 4 pages. http://www
Cox, S. M., and K. Fulsaas. 2003. Mountaineering: The freedom of the hills.
Seventh edition. The Mountaineers Books, Seattle, Washington, USA.
Cryan, P. M., C. U. Meteyer, J. G. Boyles, and D. S. Blehert. 2010. Wing
pathology of white-nose syndrome in bats suggests life-threatening
disruption of physiology. BMC Biology 8:135. http://www
.biomedc ent
Foley, J., D. Clifford, K. Castle, P. Cryan, and R. S. Ostfield. 2011.
Investigating and managing the rapid emergence of white-nose
syndrome, a novel, fatal, infectious disease of hibernating bats.
Conservation Biology 25:223–231.
Frick, W. F., J. F. Pollock, A. C. Hicks, K. E. Langwig, D. S. Reynolds, G. G.
Turner, C. M. Butchkoski, and T. H. Kunz. 2010. An emerging disease
causes regional population collapse of a common North American bat
species. Science 329:679–682.
Gargas, A., M. T. Trest, M. Christensen, T. J. Volk, and D. S. Blehert. 2009.
Geomyces destructans
sp. nov. associated with white-nose syndrome.
Myotoxin 108:147–154.
Hicks, A., C. Herzog, A. Ballmann, and A. King. 2009. Protocol for 2008–
2009 hibernacula surveys of cave bats, Indiana bat protocol. US Fish
and Wildlife Service unpublished report, Bloomington, Indiana, USA. 45
Hoyt, J. R., K. Sun, K. L. Parise, G. Lu, K. E. Langwig, T. Jiang, S. Yang, F. W.
Frick, A. M. Kilpatrick, J. T. Foster, and J. Feng. 2016. Widespread bat
white-nose syndrome fungus in northeastern China [Letter]. Emerging
Infectious Disease. doi:10.3201/eid2201.151314.
Lorch, J. M., C. U. Meteyer, M. J. Behr, J. G.Boyles, P. M. Cryan, A. C.
Hicks, A. E. Ballmann, J. T. H. Coleman, D. N. Redell, D. M. Reeder,
and D. S. Blehert. 2011. Experimental infection of bats with
causes white-nose syndrome. Nature 480(7377):376–378.
Meteyer, C. U., E. L. Buckles, D. S. Blehert, A. C. Hicks, D. E. Green, V.
Shearn-Bochsler, N. J. Thomas, A. Gargas, and M. J. Behr. 2009.
Histopathologic criteria to confirm white-nose syndrome in bats. Journal
of Veterinary Diagnostic Investigations 21:411–414.
Minnis, A. M., and D. L. Lindner. 2013. Phylogenetic evaluation of
Geomyces and allies reveals no close relatives of
, comb. nov., in bat hibernacula of eastern North America.
Fungal Biology 117:638–649.
NPS (National Park Service). 2006. Management Policies 2006. US
Department of the Interior, National Park Service. 180 pages. ht tp://
—— —. 2010. Memorandum (N16:2300) to regional directors: Revised
guidance on white-nose syndrome in bats and impacts on cave use.
National Park Service, Washington, DC. 2 pages.
Okoniewski, J. C., J. Haines, A. C. Hicks, K. E. Langwig, R. I. von Linden, and
C. A. Dobony. 2010. Detection of the conidia of
Geomyces destructans
in northeast hibernacula, at maternity colonies, and on gear—Some
findings based on microscopy and culture. (May) 2010. White-nose
Syndrome Symposium, Pittsburgh, Pennsylvania, USA.
Puechmaille, S. J., G. Wibbelt, V. Korn, H. Fuller, F. Forget, K. Mühldorfer,
A. Kurth, W. Bogdanowicz, C. Borel, T. Bosch, T. Cherezy, M. Drebet, T.
Görföl, A. J. Haarsma, F. Herhaus, G. Hallart, M. Hammer, C. Jungmann,
Y. Le Bris, L. Lutsar, M. Masing, B. Mulkens, K. Passior, M. Starrach,
A. Wojtaszewski, U. Zöphel, and E. C. Teeling. 2011. Pan-European
distribution of white-nose syndrome fungus (
Geomyces destructans
) not
associated with mass mortality. PLoS One 6 (e19167). doi:10.1371
TPWD (Texas Parks and Wildlife Department). 2017. Fungus that causes
white-nose syndrome in bats detected in Texas. News release. 23
March. Texas Parks and Widllife Department.
Turner, G. G., C. U. Meteyer, H. Barton, J. F. Gumbs, D. M. Reeder,
B. Overton, H. Bandouchova, T. Bartonicka, N. Martínková, J. Pikula,
J. Zukal, and D. S. Blehert. 2014. Nonlethal screening of bat-wing skin
with the use of ultraviolet fluorescence to detect lesions indicative of
white-nose syndrome. Journal of Wildlife Diseases 50:566–573.
Turner, G. G., D. M. Reeder, and J. T. H. Coleman. 2011. 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
USDAFS (USDA Forest Service). 2014. Cave closure order extended for
five years; forest managers hopeful a cure can be found for deadly bat
disease. Press release. Ouachita, Ozark, and St. Francis National Forests,
Hot Springs, Arkansas, USA. 2 pages.
USFWS (US Fish and Wildlife Service). 2011a. White-nose syndrome
decontamination protocol, Version 01.25.2011. US Fish and Wildlife
—— —. 2011b. Supporting decontamination documentation for cavers
(WNS decontamination supplement 1 of 2), Version 01.25.2011. US Fish
and Wildlife Service.
—— —. 2011c. Supporting decontamination documentation for
researchers (WNS decontamination supplement 2 of 2), Version
01.25.2011. US Fish and Wildlife Service.
—— —. 2012a. North American bat death toll exceeds 5.5 million from
white-nose syndrome. News release. 17 January. US Fish and Wildlife
Service. 2 pages.
—— —. 2012b. White-nose syndrome decontamination protocol, Version
03.15.2012. US Fish and Wildlife Service. 3 pages.
—— —. 2014. White-nose syndrome: The devastating disease of
hibernating bats in North America, June. 2 pages. https://www
—— —. 2015a. Fungus that causes bat disease detected in Nebraska.
News release. 12 November. 1 page. https://www.whitenosesyndrome
—— —. 2015b. US Fish and Wildlife Service protects northern long-eared
bat as threatened under Endangered Species Act. News release. 1 April.
4 pages.
—— —. 2016. National White-Nose Syndrome Decontamination Protocol,
Version 04.12.2016.
WDFW (Washington Department of Fish and Wildlife). 2016. Bat with
white-nose syndrome confirmed in Washington State. News release. 31
March. Washington Department of Fish and Wildlife. 2 pages. http://
Wibbelt, G., A. Kurth, D. Hellmann, M. Weishaar, A. Barlow, M. Veith,
J. Prüger, T. Görföl, L. Grosche, F. Bontadina, U. Zöphel, H.-P. Seidl,
P. M. Cryan, and D. S. Blehert. 2010. White-nose syndrome fungus
Geomyces destructans
) in bats, Europe. Emerging Infectious Diseases
16:12 37–1242.
Wolf, F. A., and F. T. Wolf. 1947. The fungi, Volume II. John Wiley and Sons,
New York, New York, USA.
Zukal, J., H. Bandouchova, T. Bartonicka, H. Berkova, V. Brack, J. Brichta,
M. Dolinay, K. S. Jaron, V. Kovacova, M. Kovarik, N. Martínková, K.
Ondracek, Z. Rehak, G. G. Turner, and J. Pikula. 2014. White-nose
syndrome fungus: A generalist pathogen of hibernating bats. PLOS ONE
9.5(e97224). doi:10.1371/journal.pone.0097224.
About the author
J. Judson Wynne is an assistant research professor with the
Department of Biological Sciences, Merriam-Powell Center for
Environmental Research, Northern Arizona University, Flagstaff,
Arizona 86011; website:; email: jut
Appendix I. Required equipment and explanations on use
NO GEAR used in caves within a WNS confirmed or suspected area (i.e., state) may be used in areas where the disease has not been confirmed or sus-
pected (USFWS 2016).
1. Required equipment
The amount of disinfecting supplies required depends upon team size,
number of days in the field, and number of sites visited. Generally, the
supply list provided below will accommodate a six-person team for one
week. Isopropyl alcohol (70%) wipes are a more benign decontaminant
than the other disinfectants listed in the decontamination protocol. Thus,
it would be more appropriate for backcountry work. We used Lysol®
disinfectant wipes during protocol development. Further, if caves are
characterized by walkable passage, then one pair of disposable coveralls
per person per day will be sufficient. If not, at least two pairs of ballistic
nylon coveralls per person should be considered.
A. Supply checklist for procedures prior to entry and after
exiting a cave or mine
Disposable coveralls or ballistic nylon coveralls (1 pair per person,
per study site)
Duct tape (2 large rolls)
70% isopropyl alcohol (70%) wipes (2 canisters; 44-count)
Large (10–15 gallons) heavy-duty plastic garbage bags (40-count
Plastic zip ties (25-count)
Heavy-duty resealable ziplock freezer bags (2 boxes [10-count]
each of quart and gallon sizes)
Properly laundered bandanas or rags (number depends on number
of sites visited)
Biodegradable/all-natural hand sanitizer (2 bottles; 12 oz. container)
Compressed air (two 10-ounce cans)
Trauma shears (2 pairs)
Small nylon scrub brush (1 per person)
Nitrile gloves, powder-free (2 boxes; 100 count; at least 2 pairs per
person, per day, per study site required)
Permanent black markers (1 box; 12-count)
B. Full decontamination supply checklist
Tyvek® or ProShield® coveralls (1 pair)
Tyvek® or ProShield® slip-on shoe covers (1 pair)
Disinfectant cleaner (1 gallon; refer to decontamination protocol;
USF WS 2016)
5-gallon buckets (number of buckets required depends on number
of full decontaminations required; minimum is 2 buckets per full
decontamination with the rinse bucket being reused)
2 scrub brushes, nylon-bristled
Plastic zip ties (25-count)
2 pairs rubber cleaning gloves
Biodegradable soap (16 fluid oz.)
Biodegradable wipes (2 boxes; 30-count)
C. Personal gear checklist (WNS-related)
Clothing (1 set of clothing per site; at least 1 set for transit
between study site and camp/vehicles)
Knee and elbow pads (at least 2 pairs)
Caving gloves (synthetic leather/nylon; at least 2 pairs)
1 pair PVC boots or hiking boots (number of pairs of hiking boots
required will be determined based on number of sites visited and
time required for boots to completely dry following decontamina-
1 PVC caving backpack
2 pairs gaiters (only if hiking boots are used)
Dry bags (recommended)
2. Equipment explanations and disinfecting
We provide explanations on select items listed in checklists
We also provide recommendations for disinfecting personal and project
equipment not discussed in these checklists. The instructions provided in
the most recent version of the WNS Decontamination Protocol (e.g.,
USFWS 2016) must be used for decontaminating all personal gear, equip-
ment, and clothing.
Tyvek®, ProShield®, or ballistic nylon coveralls and duct tape
Coveralls should be large enough to fit over underclothing and knee/
elbow pads. Use of coveralls will further limit contact of underclothing
and knee/elbow pads with the cave environment. In caves with walkable
passage, Tyvek® or ProShield® coveralls are appropriate. Coveralls may be
purchased with elastic cuffs on wrists, or duct tape can be used to secure
the sleeves close to the wrists. Duct tape is used to affix the pant legs of
the coveralls to boots or gaiters. In caves with constricted passageways
requiring belly crawling or moving through narrow passageways, ballistic
nylon coveralls are preferred. Upon completing work at a study site, cov-
eralls are isolated and properly stored in a heavy-duty plastic garbage bag
or gallon-sized, resealable ziplock bag.
Quaternary ammonium compounds
We used Lysol® IC Quaternary Disinfectant Cleaner. However, the WNS
Decontamination Protocol (e.g., USFWS 2016) provides several alternatives
that have been confirmed to kill
P. destructans
Nitrile gloves
Applying and removing duct tape while wearing nitrile gloves ultimately
results in torn gloves. If gloves are torn during decontamination proce-
dures, immediately wash hands and put on a new pair of gloves.
Trauma shears
For cutting duct tape from wrist cuffs and around pant legs to detach
disposable coveralls from boots/gaiters.
Editor’s Note: The following four appendixes, by J. Judson Wynne, are
intended as a “field-friendly” supplement to the preceding article, “White-
nose syndrome decontamination procedures for backcountry subterranean
projects,” also by Wynne. They can be removed from the publication,
laminated, and used in the field.
Reusing decontaminated equipment among different study sites
For equipment that requires decontaminattion by submersion in one of
the known disinfectants/applications (e.g., gloves, knee/elbow pads,
hiking boots, and gaiters), the number of extra pairs required will depend
upon the number of different study sites visited in rapid succession and
the amount of time needed for recently decontaminated equipment to
completely dr y before reuse. For most xeric regions in the southwestern
United States, one extra pair of each item (gloves, knee/elbow pads,
hiking boots, and gaiters) will probably be sufficient; the second pair may
be worn at the next study site while the recently decontaminated pair is
drying. In more mesic regions (e.g., the Pacific Northwest), drying time
may require two or more days. Workers will need to determine if (1) the
equipment will dry adequately following decontamination so it may be
reused while in the field, or (2) a pair of each item is required per study
One set of clothing per person per study site is required. These sets of
clothing will be designated for underground use only. Upon completion of
each site, clothing is isolated in a gallon-sized, resealable ziplock bag and
is properly decontaminated in the field or once field personnel have
returned to their respective homes. Additionally, a “clean” set of clothing,
which never comes into contact with dirty caving equipment or the cave
environment, is required for use in transit between the study site and
At least two pairs of synthetic leather/nylon gloves per person are
recommended. Rubber or nitrile gloves easily tear when abraded on rock
surfaces. Synthetic leather/nylon gloves are more durable and are
invaluable to work safely underground.
Knee and elbow pads
At least two sets of pads per person are recommended.
PVC boots or other footwear/gaiters
One pair of Wellington or knee-length-style PVC boots per team member
is recommended. These boots are easy to clean and dry quickly. However,
these boots often do not fit as well as hiking boots; as a result, it may be
difficult to safely navigate cave passages and scramble over boulders and
rocks. Consequently, some workers may choose to use hiking boots and
gaiters rather than PVC boots.
Cave packs
One PVC cave pack per person is recommended. It is easy to clean and
dries quickly.
One helmet, two light sources on the helmet, and at least one additional light
source within the caving bag per person are recommended. Before entering
each study site, helmets will be decontaminated with isopropyl alcohol (70%)
wipes. The use of porous headlamp straps can be eliminated by mounting the
primary light source directly on the helmet.
Miscellaneous gear
When possible, additional gear should be stored in sealed ziplock bags or
PVC dry bags within cave packs and accessed as needed. This includes
food, urine bottles, solid human waste disposal bags, medical supplies,
and tool kits. Any items used underground are properly cleaned, isolated,
or disposed of as appropriate. Water should be stored in containers (hard
plastic bottles or stainless steel containers) that can be easily disinfected.
Water bladders are not recommended because they are difficult to
Electronic sampling equipment
We recommend disinfecting electronic equipment between study sites
using (1) compressed air to carefully clean the equipment and (2) isopropyl
alcohol (70%) wipes to wipe down those areas lacking movable parts,
buttons, or screens. Isopropyl alcohol wipes are one of the cleaning agents
recommended by the WNS Decontamination Protocol (USFWS 2016).
Compressed air may help remove fungal spores/conidia from keypads and
other components. Care should be taken when wiping down electronic
equipment with alcohol wipes because some surfaces may be damaged.
Headlamps and batteries
Headlamps are disinfected by (1) removing the elastic head straps and
submersing them in solution following steps identified in the WNS
Decontamination Protocol (USFWS 2016), and (2) wiping down the
electronic parts using isopropyl alcohol (70%) wipes. Backup headlamps
and batteries should be stored in sealed individual ziplock bags, kept in
cave packs, and accessed as needed.
Vertical caving equipment
All technical caving equipment must be designated for site-specific,
regional use within a WNS confirmed, suspected, or unaffected area
(USFWS 2016). When moving between study sites within one of those
designated areas, all equipment should be cleaned following the
manufacturer’s specifications (USFWS 2016).
Exposed skin
We recommend wiping all exposed body parts with all-natural/biodegrad-
able wipes or soapy water following decontamination procedures. This is
done not as part of the decontamination protocol per se, but rather to
reduce accidental chemical exposure to any areas of skin. For example, acci-
dental exposure may occur after handling decontamination equipment,
recently decontaminated equipment, or personal gear. Users of isopropyl
alcohol wipes should follow the Centers for Disease Control and
Prevention’s occupational health guidelines (CDC 1978).
NOTE: Once all equipment and personal gear are disinfected, we
recommend using a clean bandana dampened with water or
all-natural/biodegradable wipes to remove chemical residues from
surfaces. This will reduce chemical exposure of field personnel.
Appendix II. Fieldwork preparations
NO GEAR used in caves within a WNS confirmed or suspected area (i.e., state) may be used for subterranean research in those states where the disease
has not been confirmed or suspected (USFWS 2016).
1. Packing decontamination supplies to and from study
Options for portaging decontamination supplies:
There are two general
approaches that may be undertaken for mobilizing decontamination
equipment at a study site. (1) For study sites within 1 mile of vehicles/
camp, workers may choose to portage multiple containers and required
equipment itemized in Appendix I directly to the study site.
Decontamination equipment and disinfectant containers used during
decontamination procedures must be cleaned with isopropyl alcohol
(70%) wipes (USFWS 2016) before these materials are integrated with
other equipment and packed out. (2) Regarding single study sites
requiring a many-mile hike to access a particular backcountry site, it may
be easier for each person on the backcountry team to have a personal
decontamination kit with all of the necessary items to follow the WNS
Decontamination Protocol (USFWS 2016) in the field; they will also be
responsible for packing their own personal supplies into and out of the
remote study site. Personal decontamination kits should contain the
following: 16 isopropyl alcohol (70%) wipes, eight all natural/
biodegradable wipes, personal hand sanitizer, two pairs of nitrile gloves,
two garbage bags, and two zip ties. Supplies must be multiplied
appropriately if more than one site is to be visited in a given day. Two
pairs of trauma shears are adequate for a six-person team and may be
carried to and from the site by a designated person.
2. Personal equipment
Personal equipment should be assembled as follows: (1) One pair of
disposable or ballistic nylon coveralls per person per day. For visiting
multiple study sites in a given day, the number of coveralls will increase
accordingly. (2) Cache of personal duct tape per person to be used to tape
down wrist cuffs, secure boots/gaiters to coveralls, and repair disposable
coveralls as necessary. (3) Various-sized disposable ziplock freezer bags
(e.g., sandwich, quart, and gallon sizes). The number of bags per size
depends upon the equipment required for a specific research task and the
needs of each team member. We recommend double bagging all
3. Establishing staging areas at the study site
Three staging areas near the study site entrance should be established
and designated as “clean,” “decontamination,” and “intermediate”
zones. All zones should be at least 20 m (66 ft) apart, and clean and
intermediate zones must be located upwind from the decontamination
zone. (1) The clean zone is used to stage non-cave-related gear (e.g.,
backpacks, extra water bottles, satellite phone, and other equipment),
and to change into clean coveralls, boots, and other equipment. (2) The
decontamination zone is where chemical disinfectants, isopropyl
alcohol and all-natural/biodegradable wipes, hand sanitizer, trauma
shears, garbage bags, zip ties, and related supplies are staged. It is also
the location for disinfecting equipment, cleaning exposed parts of the
body, stripping off and isolating coveralls, and changing into clean
clothing. If logistics permit, a hand and body washing station may be
established where personnel may clean themselves prior to returning to
the clean zone. (3) The intermediate zone must be established in an
area that team members do not have to walk through to reach the clean
zone. This area is used to stage clean boots and a clean change of clothes
(isolated in a ziplock bag for the hike back to vehicles/camp). This zone is
also used for staging recently cleaned gear to be moved into the clean
zone once decontamination procedures are completed. Refer to the
following protocols for clarification on zones and their functions.
4. Establishing an area for full decontamination/
disinfecting of equipment prior to changing study sites
Designate a decontamination area at least 20 m (66 ft) downwind from
vehicles and camp. All decontamination and containment supplies, and
personal isolation bags (i.e., gear to be disinfected) should be placed
within this area. Full decontamination of equipment (e.g., caving bags,
PVC or hiking boots and gaiters, knee/elbow pads, gloves, and any other
gear that requires submersion in a chemical mixture) between individual
study sites should be determined at the discretion of the jurisdictional
regulatory or resource management agency.
5. General recommendations and notes
Proper disposal of camp refuse: Clearly label and segregate garbage
bags designated for “contaminated” items from daily camp/project
Proper storage of duct tape: Do not place inexpensive duct tape in
direct sunlight during warm spring and summer months or store in hot
areas (e.g., enclosed vehicles). Depending on the type of duct tape
used, the glue adhesive may melt. Gorilla® duct tape works well when
exposed to direct sunlight and heat.
After exiting a study site, decontaminate equipment (helmets, water
bottles, urine bottles, metal clipboards, cave survey equipment, and
the exterior ziplock and dry bags containing gear) using a three-step
procedure. (1) Physically remove dirt and mud from boots, coveralls,
and caving and other equipment using nylon brushes. (2) Carefully
clean surfaces with isopropyl alcohol (70%) wipes. (3) Remove
chemical residue by wiping down all surfaces with all-natural/
biodegradable wipes or a clean, damp bandana. This should be done
prior to removing gloves.
Team members should work together and watch each other to ensure
appropriate decontamination protocols are applied between study
sites. We recommend using the “buddy system” when possible. With
team members watching one another, this will reduce the likelihood of
overlooking gear, equipment, and clothing that require disinfecting.
When working in caves characterized by narrow passages or sections
requiring belly crawls, “buddies” are responsible for periodically
inspecting each other’s coveralls for breaches and work together
during decontamination to make sure all steps are being followed and
decontamination proceeds correctly.
If personnel are portaging equipment into and out of a remote area,
quart- and gallon-sized ziplock bags may be used to store used
coveralls, nitrile gloves, disinfecting wipes, and other gear. Garbage
bags are required to store PVC backpacks and boots. These items
should then be placed within a larger backpack for hiking to and from
remote study sites. However, compartmentalizing smaller items and
equipment in smaller resealable ziplock bags may make packing
equipment into a backpack much easier.
For large multiperson projects, individuals should label their personal
isolation bag containing their cave bag, boots, and other equipment
by writing their name on a strip of tape adhered to their bag.
When personnel spend multiple days at the same study site, it may be
easier to stage most of the personal and WNS disinfection equipment
in the appropriate zones—provided that the site is secure and the risk
of theft is low.
When staging caving equipment at a study site overnight, properly
secure all equipment to prevent entry of insects and rodents. Be
certain to remove any food items from gear that will be left overnight.
Staff with regulatory or resource management agencies may require
cave/mine personnel to bathe upon completion of operations at each
study site prior to returning to their vehicles/camp (e.g., P. Ormsbee,
2011, personal communication).
Print all field forms on weather-/waterproof paper for easier
decontamination upon return to the office.
Appendix III. Procedures prior to entering and after exiting a cave or mine
This approach discusses preparations for daily decontamination activities, including procedures prior to and after exiting a study site.
1. Prior to entering a cave or mine
Multiday or single visits to a study site:
(1) Upon arrival at a study site,
stage non-caving gear (e.g., backpacks, hats, and trekking poles) in the
clean zone. (2) Put on caving clothes and knee/elbow pads, then clean
coveralls. (3) If coveralls lack elastic wrist cuffs, use duct tape to secure
wrist cuffs to prevent the coverall arms from sliding up and exposing skin
or underclothing.1 (4) Put on boots. (5) Tape pant legs of coveralls to each
boot/gaiter.2 (6) For additional protection against a disposable coverall
breach, place duct tape on elbows and knees (and seat, if necessary). (7)
Stage clean clothing and hiking boots in the intermediate zone and place
disinfecting equipment in the decontamination zone.
Return to same study site for multiple days at a cave/mine:
(1) Stage non-
caving equipment in the clean zone. (2) Proceed to the decontamination
zone to obtain cave clothing, knee/elbow pads, boots, and related gear.3
(3) Carefully remove clean clothing and place it in a clean, resealable
ziplock bag. (4) Remove cave clothing from the sealed ziplock bag. (5)
Follow steps 2 through 7 above (“Multiday or single visits to a study
2. Procedures after exiting a cave or mine
(1) After exiting the study site, proceed directly to the decontamination
zone. (2) Put on nitrile gloves. (3) Isolate field forms and notes.4 (4) Brush
off boots with nylon brush and disinfect coveralls and boots using
isopropyl alcohol (70%) wipes. (5) Remove all necessary equipment from
caving bag and place caving bag in the personal isolation bag (i.e., a
garbage bag).5 (6) Disinfect all personal equipment (e.g., helmet, water
bottles, urine bottles, dry bags containing vertical gear and the exterior of
ziplock bags used for equipment and isolating field forms) and group gear
(e.g., metal clipboards, cave survey equipment, and electronics) using the
most appropriate decontamination procedures (e.g., the three-step
procedure for all equipment except vertical gear6). Disinfect vertical gear
1 In some cases it may be possible to tape down glove cuffs to the coverall cuffs. However,
from our experience, it is necessary to remove gloves to use survey and electronic equipment.
Additionally, when working several hours at a given study site, this may also be impractical for
2 Before taping, be sure to slide pant leg cuff up boot approximately 5 cm (2 in) to permit
knee-bending mobility. If using hiking boots and gaiters, tape the top of the gaiter to the
coveralls and the bottom of the gaiter to the boot.
3 All equipment will have been staged at the study site on the previous day. Change from clean
clothing into cave clothing and coveralls in the decontamination zone.
4 Access to data collected daily is important, so that it can be evaluated and logged each
evening. Place each page in its own resealable ziplock bag. Do not place multiple pages in one
bag, because it cannot be opened at camp without risk of contamination.
5 If operations at the study site are complete, dump contents of the caving bag onto the ground
and place the caving bag in a garbage bag. If returning to the study site the next day, remove
equipment from the caving bag that needs to be maintained for the next day (e.g., water bot-
tles for replenishing, batteries for recharging, urine bottles for cleaning). In either case, follow
the most recent WNS Decontamination Protocol (e.g., USFWS 2012).
6 Decontaminate using isopropyl alcohol (70%) wipes. For water bottles, rinse the exterior with
clean water prior to reuse.
in accordance with the most recent WNS Decontamination Protocol (e.g.,
USFWS 2016). (7) Clean gloved hands.6 (8) Move disinfected equipment to
the intermediate zone and retrieve clean clothing and hiking boots. (9)
Return to the decontamination zone and use trauma shears to cut the
duct tape wrapped around wrist cuffs and PVC boots/gaiters. (10) Remove
PVC boots/hiking boots and then coveralls by turning the suit inside out.
(11) Step into the inside of the coveralls. If there are no ruptures, the
inside of your coveralls should not be “contaminated.” Therefore, you can
safely stand on them, keeping your socks clean, until they are isolated in
the subsequent steps. (12) Place PVC boots/hiking boots and knee/elbow
pads in the personal isolation bag along with the previously isolated
caving bag. (13) Remove clothing worn under the coveralls, place in a
resealable ziplock bag, and deposit in the personal isolation bag. (14)
Clean gloved hands and personal isolation bag.6 (15) Wipe down the
inside of the bag up to where it is tied or zip-tied.7 (16) Wipe personal
isolation bag with isopropyl alcohol wipes followed by all-natural/
biodegradable wipes. (17) Using all-natural/biodegradable wipes, clean
exposed areas of skin (e.g., face, neck, and arms) and areas exposed to
isopropyl alcohol during decontamination procedures.7 (18) Change into
clean clothes. (19) Step out of coveralls when changing into clean boots.
(20) With gloved hands, carefully place used coveralls into the appropriate
isolation bag (e.g., personal or group) for either laundering/
decontamination or disposal. (21) Remove nitrile gloves using standard
medical glove removal procedures.8 (22) Place used nitrile gloves into the
appropriate isolation bag. (23) Clean hands. (24) Tie or zip-tie the group
and personal isolation bags.9 (25) Retrieve disinfected equipment from the
intermediate zone and proceed to the clean zone.
7 If logistics permit, biodegradable soap and water should be used to wash hands and body. P.
Ormsbee (2011, personal communication) used this approach in Washington State. By vigor-
ously washing hands and exposed parts of the body with soap and water, you can potentially
P. destructans
hyphae and spores/conidia mechanically. If this is not possible, use
antibacterial hand sanitizer and antibacterial wipes in an attempt to mechanically remove the
hyphae and conidia from skin.
8 Pinch glove of one hand carefully on the inside of the wrist and remove it by turning it inside
out, then remove the other glove using the clean interior of the glove previously removed.
9 The last person to complete the procedures after exiting a cave or mine is responsible for
closing the group isolation bag. Follow same procedures as for closing personal isolation bags.
Appendix IV. Procedures for full decontamination
Another option is to designate one person to be responsible for
disinfecting all personal equipment, as follows.
1. Put on a clean pair of disposable or ballistic nylon coveralls, shoe
covers, and rubber cleaning gloves.
2. Prepare chemical decontamination mixture (see USFWS 2016) in
5-gallon bucket1 and clean water for rinsing (in another 5-gallon
3. Dump all equipment out of personal isolation garbage bags and place
all empty personal isolation bags in a clean garbage bag.
4. Use a nylon brush to remove any dirt and mud from boots, caving
bags, knee/elbow pads, gloves, and other equipment (USFWS 2016).
5. Decontaminate gear following the most recent WNS Decontamination
Protocol (e.g., USFWS 2016). Submerge recently disinfected gear in
rinse water following decontamination treatment.
6. Once all gear is disinfected, nylon brushes and any other equipment
used in the decontamination process are decontaminated following
step 5.
7. Remove rubber gloves and put on a pair of nitrile gloves.
8. Decontaminate rubber gloves.2
9. Carefully roll up pant legs of coveralls so they do not touch shoe
10. Remove shoe covers and place in a garbage bag (same bag used to
dispose of personal isolation bags).
11. Remove shoes (the ones worn underneath the shoe covers) and place
on ground in front of you.
12. Remove coveralls by peeling them off and turning the suit inside out.
13. Put on shoes.
14. Place coveralls in garbage bag.
15. Wipe exterior of garbage bag. Wipe inside of bag to area below where
zip tie will be secured.
16. Remove nitrile gloves following standard medical procedure.3
17. Place nitrile gloves in garbage bag.
18. Close and seal the garbage bag with a zip tie.
19. Double bag the garbage bags containing contaminated or presumed
contaminated gear/garbage and zip tie shut.
20. Wash hands with all-natural/biodegradable wipes or soap and water.
1 One mixture of decontamination solution is prepared in a 5-gallon bucket and used on a per
study site basis. Used solution remains in the bucket, covered with a tightly fastened lid, and
is then placed in a garbage bag and zip-tied. The bucket is stored securely in the vehicle to
prevent it from tipping and spilling while driving on rough roads. All chemical-water mixtures
are properly disposed of following the manufacturer’s recommendations at the nearest suitable
facilities exist.
2 Decontaminate using isopropyl alcohol (70%) wipes.
3 Pinch glove of one hand carefully on the inside of the wrist and remove it by turning it inside
out, then remove the other glove using the clean interior of the glove previously removed.
Literature cited
CDC (Centers for Disease Control and Prevention). 1978. Occupational
health guidelines for isopropyl alcohol.
Ormsbee, P. 2011. Personal communication: E-mail to J. Judson Wynne, 16
November. USDA Forest Service Region 6, Portland, Oregon, USA.
USFWS (US Fish and Wildlife Service). 2016. National white-nose
syndrome decontamination protocol, Version 04.12.2016. https://
Full-text available
Since the initial experiments nearly 50 years ago, techniques for detecting caves using airborne and spacecraft acquired thermal imagery have improved markedly. These advances are largely due to a combination of higher instrument sensitivity, modern computing systems, and processor intensive analytical techniques. Through applying these advancements, our goals were to: (1) Determine the efficacy of methods designed for terrain analysis and applied to thermal imagery; (2) evaluate the usefulness of predawn and midday imagery for detecting caves; and (3) ascertain which imagery type (predawn, midday, or the difference between those two times) was most informative. Using forward stepwise logistic (FSL) and Least Absolute Shrinkage and Selection Operator (LASSO) regression analyses for model selection, and a thermal imagery dataset acquired from the Mojave Desert, California, we examined the efficacy of three well-known terrain descriptors (i.e., slope, topographic position index (TPI), and curvature) on thermal imagery for cave detection. We also included the actual, untransformed thermal DN values (hereafter "unenhanced thermal") as a fourth dataset. Thereafter, we compared the thermal signatures of known cave entrances to all non-cave surface locations. We determined these terrain-based analytical methods, which described the "shape" of the thermal landscape hold significant promise for cave detection. All imagery types produced similar results. Down-selected covariates per imagery type, based upon the FSL models, were: Predawn-slope, TPI, curvature at 0 m from cave entrance, as well as slope at 1 m from cave entrance; midday-slope, TPI, and unenhanced thermal at 0 m from cave entrance; and difference-TPI and slope at 0 m from cave entrance, as well as unenhanced thermal and TPI at 3.5 m from cave entrance. Finally, we provide recommendations for future research directions in terrestrial and planetary cave detection using thermal imagery.
Full-text available
We describe and illustrate the new species Geomyces destructans. Bats infected with this fungus present with powdery conidia and hyphae on their muzzles, wing membranes, and/or pinnae, leading to description of the accompanying disease as white-nose syndrome, a cause of widespread mortality among hibernating bats in the northeastern US. Based on rRNA gene sequence (ITS and SSU) characters the fungus is placed in the genus Geomyces, yet its distinctive asymmetrically curved conidia are unlike those of any described Geomyces species.
Full-text available
White-nose syndrome (WNS) has caused recent catastrophic declines among multiple species of bats in eastern North America. The disease's name derives from a visually apparent white growth of the newly discovered fungus Geomyces destructans on the skin (including the muzzle) of hibernating bats. Colonization of skin by this fungus is associated with characteristic cutaneous lesions that are the only consistent pathological finding related to WNS. However, the role of G. destructans in WNS remains controversial because evidence to implicate the fungus as the primary cause of this disease is lacking. The debate is fuelled, in part, by the assumption that fungal infections in mammals are most commonly associated with immune system dysfunction. Additionally, the recent discovery that G. destructans commonly colonizes the skin of bats of Europe, where no unusual bat mortality events have been reported, has generated further speculation that the fungus is an opportunistic pathogen and that other unidentified factors are the primary cause of WNS. Here we demonstrate that exposure of healthy little brown bats (Myotis lucifugus) to pure cultures of G. destructans causes WNS. Live G. destructans was subsequently cultured from diseased bats, successfully fulfilling established criteria for the determination of G. destructans as a primary pathogen. We also confirmed that WNS can be transmitted from infected bats to healthy bats through direct contact. Our results provide the first direct evidence that G. destructans is the causal agent of WNS and that the recent emergence of WNS in North America may represent translocation of the fungus to a region with a naive population of animals. Demonstration of causality is an instrumental step in elucidating the pathogenesis and epidemiology of WNS and in guiding management actions to preserve bat populations against the novel threat posed by this devastating infectious disease.
Full-text available
White-nose syndrome (WNS) is causing unprecedented declines in several species of North American bats. The characteristic lesions of WNS are caused by the fungus Geomyces destructans, which erodes and replaces the living skin of bats while they hibernate. It is unknown how this infection kills the bats. We review here the unique physiological importance of wings to hibernating bats in relation to the damage caused by G. destructans and propose that mortality is caused by catastrophic disruption of wing-dependent physiological functions. Mechanisms of disease associated with G. destructans seem specific to hibernating bats and are most analogous to disease caused by chytrid fungus in amphibians.
Full-text available
White-nose syndrome (WNS) is an emerging disease affecting hibernating bats in eastern North America that causes mass mortality and precipitous population declines in winter hibernacula. First discovered in 2006 in New York State, WNS is spreading rapidly across eastern North America and currently affects seven species. Mortality associated with WNS is causing a regional population collapse and is predicted to lead to regional extinction of the little brown myotis (Myotis lucifugus), previously one of the most common bat species in North America. Novel diseases can have serious impacts on naïve wildlife populations, which in turn can have substantial impacts on ecosystem integrity.
Full-text available
White-nose syndrome (WNS) is a cutaneous fungal disease of hibernating bats associated with a novel Geomyces sp. fungus. Currently, confirmation of WNS requires histopathologic examination. Invasion of living tissue distinguishes this fungal infection from those caused by conventional transmissible dermatophytes. Although fungal hyphae penetrate the connective tissue of glabrous skin and muzzle, there is typically no cellular inflammatory response in hibernating bats. Preferred tissue samples to diagnose this fungal infection are rostral muzzle with nose and wing membrane fixed in 10% neutral buffered formalin. To optimize detection, the muzzle is trimmed longitudinally, the wing membrane is rolled, and multiple cross-sections are embedded to increase the surface area examined. Periodic acid-Schiff stain is essential to discriminate the nonpigmented fungal hyphae and conidia. Fungal hyphae form cup-like epidermal erosions and ulcers in the wing membrane and pinna with involvement of underlying connective tissue. In addition, fungal hyphae are present in hair follicles and in sebaceous and apocrine glands of the muzzle with invasion of tissue surrounding adnexa. Fungal hyphae in tissues are branching and septate, but the diameter and shape of the hyphae may vary from parallel walls measuring 2 microm in diameter to irregular walls measuring 3-5 microm in diameter. When present on short aerial hyphae, curved conidia are approximately 2.5 microm wide and 7.5 microm in curved length. Conidia have a more deeply basophilic center, and one or both ends are usually blunt. Although WNS is a disease of hibernating bats, severe wing damage due to fungal hyphae may be seen in bats that have recently emerged from hibernation. These recently emerged bats also have a robust suppurative inflammatory response.
Full-text available
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 psychrophilic 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.
White-nose syndrome (WNS) is a fatal disease of bats that hibernate. The etiologic agent of WNS is the fungus Geomyces destructans, which infects the skin and wing membranes. Over 1 million bats in six species in eastern North America have died from WNS since 2006, and as a result several species of bats may become endangered or extinct. Information is lacking on the pathogenesis of G. destructans and WNS, WNS transmission and maintenance, individual and site factors that contribute to the probability of an outbreak of WNS, and spatial dynamics of WNS spread in North America. We considered how descriptive and analytical epidemiology could be used to fill these information gaps, including a four-step (modified) outbreak investigation, application of a set of criteria (Hill's) for assessing causation, compartment models of disease dynamics, and spatial modeling. We cataloged and critiqued adaptive-management options that have been either previously proposed for WNS or were helpful in addressing other emerging diseases of wild animals. These include an ongoing program of prospective surveillance of bats and hibernacula for WNS, treatment of individual bats, increasing population resistance to WNS (through vaccines, immunomodulators, or other methods), improving probability of survival from starvation and dehydration associated with WNS, modifying hibernacula environments to eliminate G. destructans, culling individuals or populations, controlling anthropogenic spread of WNS, conserving genetic diversity of bats, and educating the public about bats and bat conservation issues associated with WNS.
Mountaineering: The freedom of the hills. Seventh edition. The Mountaineers Books
  • S M Cox
  • K Fulsaas
Cox, S. M., and K. Fulsaas. 2003. Mountaineering: The freedom of the hills. Seventh edition. The Mountaineers Books, Seattle, Washington, USA.