Detection of the parasite Ribeiroia ondatrae in water bodies and possible impacts of malformations in a frog host

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dc.contributor.authorHuver, Johannes Richard
dc.contributor.examiningcommitteeHubner, Erwin (Biological Sciences) Johnson, Pieter (Ecology and Evolutionary Biology, University of Colorado)en_US
dc.contributor.supervisorWhyard, Steve (Biological Sciences) Koprivnikar, Janet (Biological Sciences)en_US
dc.date.accessioned2013-07-04T13:08:30Z
dc.date.available2013-07-04T13:08:30Z
dc.date.issued2013-07-04
dc.degree.disciplineBiological Sciencesen_US
dc.degree.levelMaster of Science (M.Sc.)en_US
dc.description.abstractThis study devised a method to detect Ribeiroia ondatrae (class Trematoda) in water-bodies using environmental DNA (eDNA) collected from filtered water samples from selected ponds in the USA and Canada. Species-specific PCR primers were designed to target the Internal Transcribed Spacer-2 (ITS-2) region of the parasite’s genome. The qualitative PCR method was 70% (n=10) accurate in detecting R. ondatrae in ponds previously found to contain the parasite, while the qPCR method was 88.9% (n=9). To examine how the retinoic acid (RA) pathway gene expression may be perturbed during R. ondatrae infections, leading to limb development abnormalities in the wood frog (Lithobates sylvaticus). Multiple sequence alignments were used to design degenerate PCR primers to eight RA biosynthesis genes, but only two gene fragments were identified using this approach. Without effective primer sets it was not possible to measure changes in gene expression in infected frogs.en_US
dc.description.noteOctober 2013en_US
dc.identifier.urihttp://hdl.handle.net/1993/21697
dc.language.isoengen_US
dc.rightsopen accessen_US
dc.subjecteDNAen_US
dc.subjectRibeiroiaen_US
dc.subjectondatraeen_US
dc.subjectAnuranen_US
dc.subjectFrogen_US
dc.subjectmalformationen_US
dc.subjectLimben_US
dc.subjectdeformitiesen_US
dc.subjectdetectionen_US
dc.subjectamphibianen_US
dc.subjectTrematodaen_US
dc.subjectRAen_US
dc.subjectretinoidsen_US
dc.subjectretinoicen_US
dc.subjectITS-2en_US
dc.subjectfiltrationen_US
dc.subjectDNAen_US
dc.subjectLithobatesen_US
dc.subjectsylvaticusen_US
dc.subjectWoodfrogen_US
dc.titleDetection of the parasite Ribeiroia ondatrae in water bodies and possible impacts of malformations in a frog hosten_US
dc.typemaster thesisen_US
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