Testing for the Presence of the Chytrid fungus (Batrachochytrium Dendrobatidis) in Amphibian Populations Across Nebraska

Many worldwide amphibian population declines and mass mortality events have been attributed to a fungal infection, chytridiomycosis (chytrid), caused by the fungal zoospore Batrachochytrium dendrobatidis (Bd). Although the exact mechanism by which mass mortalities occur from chytrid is unknown, it is hypothesized that the chytrid fungus infects keratinized epidermal cells of post-metamorphic frogs with death caused by: 1) disruption of osmoregulation, 2) the absorption of a fungal toxin, 3) or a combination of these factors. Concern over the potential ecological consequences of such rapid and drastic extinctions has led to an increase in effort studying the potential effects of emerging infectious disease on amphibian populations. Furthermore, scientific and technological advances in non-invasive techniques to detect the chytrid fungus have changed the ability of researchers and managers to track the distribution of and measure the population fluctuations and declines caused by infectious disease such as chytrid.

The chytrid fungus is known to occur in Nebraska and has been found in amphibian populations located in eastern Nebraska as well as along the Central Platte River. Although sporadic testing for the chytrid fungus in populations of native amphibians has occurred in Nebraska, a statewide survey has never been conducted. This gap in our knowledge pertaining to the current distribution of chytrid complicates management and conservation of amphibians. Filling this gap will allow researchers to know where chytrid is currently found in the state, and will aid in the development of predictive models and help in the understanding of factors that may help or mitigate the further spread of chytrid.


To determine the current extent of chytrid in Nebraska by swabbing larval amphibian populations statewide. Using PCR, the samples were tested for the presence of Bd zoospores. The presence/absence of chytrid in amphibian populations were used to model the distribution of chytrid based on environmental covariates associated with wetland condition and amphibian call surveys. The modelling results will be used to develop predictive maps of the potential spread of chytrid based on important environmental and anthropogenic variables.

Current Status

During the spring and summer of 2011 and 2012, researchers visited wetlands in eastern and central Nebraska. During site visits, tadpoles were captured using dip nets. At those sites where tadpoles were captured, each individual was swabbed using a sterile swab. In 2011 and 2012, 168 swabs were collected from five frog species at 21 sites. Preliminary PCR results detected chytrid in 62% of the sites. Additionally, chytrid was detected in at least one swab from each species. The detection rate (d) was highest in bullfrogs (n = 9; d = 1.00) and lowest in plains leopard frogs (n = 78; d = 0.10). The detection rate did not appear to vary between months (May -€“ n = 56, d = 0.125; June-€“ n = 112 d = 0.17). An additional ten samples were collected during 2013 and 23 samples were collected during 2014.

Principal Investigator(s)
-Craig R. Allen, NE CFWRU
-Ted LaGrange, Nebraska Game and Parks Commission
Graduate Student(s)
-Nick Smeenk, Ph.D.
Project Duration
August 2010- May 2015
-Nebraska Game and Parks Commission
-U.S. Environmental Protection Agency (EPA)
Project Location
Statewide Nebraska