National Reconnaissance of Wetland Habitats to Determine the Presence of the Amphibian Fungus Batrachochytrium salamandrivorans (Bsal).-- in Great Smoky Mountains National Park.

Calhoun, Daniel

Field Methods: Eastern red-spotted newts will be captured by hand, minnow trap, and dipnet along the perimeter of the wetland. Individuals will be handled using disposable single-use vinyl gloves, placed in single-use plastic zip-top bags, swabbed, and released at point of capture. Up to three visits may occur throughout the activity season (March through September) at a single wetland to obtain the desired number of samples. Swabs will stored frozen until submission to the National Wildlife Health Center for PCR analysis. Any species observed at a wetland exhibiting obvious signs of illness may also be collected (maximum 5 individuals at a single wetland) for immediate submission to the USGS National Wildlife Health Center for diagnostic examination following standard NWHC SOPs: Live larvae will be placed in heavy plastic bags or plastic containers that can be secured closed with snap top or screwtop lid; the container will contain equal volumes of air and water to assure adequate oxygen exchange. Live adult or terrestrial amphibians will be placed in plastic boxes with wet unbleached brown paper towels or wet local vegetation, and three or more small holes will be made in the lid of the container to ensure adequate oxygen exchange. All individuals within containers will be transported in coolers to minimize stress and decrease metabolic demands. Deceased individuals will be double-bagged and frozen until submission to the National Wildlife Health Center. All equipment will be cleaned and disinfected between wetlands with a 10% bleach or comparable solution to minimize contamination and spread of disease among locations (following NEPARC disinfection protocol, available at: http://northeastparc.org/disinfection-protocol/). Approval for collection of data, handling, and collection of animals has been obtained from the Patuxent Wildlife Research Center Animal Care and Use Committee (ACUC).

National Reconnaissance of Wetland Habitats to Determine the Presence of the Amphibian Fungus Batrachochytrium salamandrivorans (Bsal).-- in Great Smoky Mountains National Park.

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