DETECTION AND DISTRIBUTION OF THE AMPHIBIAN FUNGAL DISEASE CHYTRIDIOMYCOSIS IN PERUVIAN AMPHIBIANS
Kosch, Tiffany A.
Chytridiomycosis is an amphibian disease caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd;Longcore et al. 1999). This disease has been identified by the Amphibian Conservation Action Plan as one of the main causal agents of worldwide amphibian declines and extinctions, and as the worst infectious disease ever recorded among vertebrates. Prior to this investigation, little was known about the prevalence of Bd across Peru, a country renowned for its high amphibian diversity and endemism. Here, I report the results of a sampling effort for Bd prevalence from the dry seasons of 2007 and 2008, which showed that among site Bd prevalence ranged from 0 to 25%. In this study, Bd was detected in 11 of 983 individuals sampled, and in nine out of 38 sites ranging in altitude from 96-3240 meters. I also discuss the implications of these results and suggest directions for further studies of Bd in Peru in order to better understand the ecology of this devastating disease and its effects on this biodiverse region. In addition to investigating the prevalence of Bd throughout Peru, I also was interested in comparing and developing methods for the detection of Bd. Since the discovery of Bd, several methods have been utilized for detection; among these PCR from skin swabs is accepted as the best method due to its high sensitivity, non-invasiveness and ease of use. However, since PCR is a chemical reaction that requires a specific DNA template to proceed, this method relies upon both the presence of non-degraded DNA template and reaction components that do not inhibit the process. Here I present the results of experimental comparisons of techniques for sample preservation, DNA extraction, and PCR. My results show that the most advantageous techniques for a Bd-field study are swab preservation in 95% ethanol, DNA extraction with DNeasy, and an end-point PCR disease assay with BSA and Amplitaq Gold. I also recommend the use of PowerClean for samples with suspected PCR inhibitors. My hope is that these techniques will promote increases in sample collection and laboratory analyses for Bd research in developing countries where such activities are drastically needed.
Kosch, Tiffany A.. (January 2012). DETECTION AND DISTRIBUTION OF THE AMPHIBIAN FUNGAL DISEASE CHYTRIDIOMYCOSIS IN PERUVIAN AMPHIBIANS (Doctoral Dissertation, East Carolina University). Retrieved from the Scholarship. (http://hdl.handle.net/10342/4023.)
Kosch, Tiffany A.. DETECTION AND DISTRIBUTION OF THE AMPHIBIAN FUNGAL DISEASE CHYTRIDIOMYCOSIS IN PERUVIAN AMPHIBIANS. Doctoral Dissertation. East Carolina University, January 2012. The Scholarship. http://hdl.handle.net/10342/4023. June 25, 2018.
Kosch, Tiffany A., “DETECTION AND DISTRIBUTION OF THE AMPHIBIAN FUNGAL DISEASE CHYTRIDIOMYCOSIS IN PERUVIAN AMPHIBIANS” (Doctoral Dissertation., East Carolina University, January 2012).
Kosch, Tiffany A.. DETECTION AND DISTRIBUTION OF THE AMPHIBIAN FUNGAL DISEASE CHYTRIDIOMYCOSIS IN PERUVIAN AMPHIBIANS [Doctoral Dissertation]. Greenville, NC: East Carolina University; January 2012.
East Carolina University