White-nose syndrome decontamination procedures for backcountry subterranean projects

By J. Judson Wynne

White-nose syndrome (WNS) has resulted in the mortalities of more than five million bats (USFWS 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 lucifugus), 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 septentrionalis; USFWS 2015b). In Canada, the Canadian Cooperative Wildlife Health Centre emergency-listed the tricolored (Perimyotis subflavus), little brown (Myotis lucifugus), and northern 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). Because 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).

A: Cluster of healthy Townsend's big-eared bats. B: Cluster of healthy Townsend's big-eared bats. C: A tricolored bat in a cave at Cloudland State Park, Georgia, 2013, with fully expressed white-nose syndrome. D: A little brown bat at the Greeley Mine, Ve
Figure 1. (Left two images) Two clusters of healthy hibernating Townsend’s big-eared bats at Grand Canyon–Parashant National Monument, Arizona. Examples of (third from left) a tricolored bat in a cave at Cloudland State Park, Georgia, 2013, and (far right) a little brown bat at the Greeley Mine, Vermont, 2009, with fully expressed white-nose syndrome.

(Left two): Courtesy of J. Judson Wynne, Northern Arizona University; (third from left): Courtesy of Pete Pattavina; (far right): Courtesy of Marvin Moriarty

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, southward 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 expansion into Minnesota, Nebraska, and Texas. Updated maps of the spread are maintained at https://www.whitenosesyndrome.org.

Map of the eastern United States and the state of Washington showing the distribution of white-nose syndrome by county and year, and whether confirmed (black outlines) or suspected (blue outlines).
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 the linked table.)

Source: Lindsey Heffernan, Pennsylvania Game Commission; Updated and adapted by NPS

The primary vector believed responsible for the westward expansion 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 swarming 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 humans. WNS has been identified in 13 bat species from cave hibernacula 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 immediately 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 epizootic 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 eastern 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 developing 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 provided a protocol for WNS decontamination (e.g., USFWS 2016) for regions where the disease is confirmed, suspected, or unconfirmed. 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 requirements 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 environmental 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 decontamination 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 submerse 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 decontamination protocols (USFWS 2012b), NPS resource managers, 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 limited. Ten different field personnel tested and refined these techniques 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 procedures 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 debriefing whereby problem areas with applying these procedures were captured and improvements were made accordingly. Additionally, we compared DuPont™ brand Tyvek® and ProShield® model disposable coveralls specifically for durability in constricted passageways 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 Personnel Involved1 Procedures Applied2 Total3
March 2011 3 11 33
June 2011 7 5 35
September 2011 5 4 20
March 2012 4 3 12
Total 121 23 100
Note: The procedures were tested and developed at Grand Canyon–Parashant National Monument, Arizona.
1Several of the same team members participated on multiple trips; therefore, the total number provided is for the number of individuals who participated in this work.
2The number of times each team applied the procedures.
3The total number of times procedures were applied per trip, calculated by the number of personnel times the number of procedures used in the field.


Through rigorous field testing, we developed a set of stepwise procedures for disinfecting field equipment and provide recommendations 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 procedures for full decontamination (i.e., prior to moving from one study site to another). The appendixes follow this article.

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-containment-zone approach (Appendix II, Section 3). The three containment areas are the (1) clean zone, an area 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 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 vehicles/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 abrading 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 environment. 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 becoming 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 decontaminating clothing and equipment. Although they were developed 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 improved, or when additional information prompts further revision. One method for improving these techniques may be through working with professionals outside the disciplines of microbiology 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 decontaminate equipment and personnel and thus better protect bat populations.

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, addenda, 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 appendixes provide a stepwise approach for decontaminating equipment and personnel (in compliance with the WNS Decontamination Protocol), there are limitations. For many cave research projects, workers must use expensive, often irreplaceable electronic 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 Decontamination 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 breaches, 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 passageways. 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 designation 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 demonstrate 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 following 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 possible to use ropes, webbing, harnesses, and other vertical gear and rope rigging equipment with soft components at different sites after cleaning this equipment following the manufacturer’s recommendations. 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 confirmed 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 decontamination 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 paradigm.


The author extends much gratitude to Jeff Bradybaugh (formally of Grand Canyon–Parashant National Monument) for supporting 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 procedure 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 Conservation 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 between 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.

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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: http://www.jutwynne.com; e-mail: jut.wynne@nau.edu.

Last updated: July 31, 2017