Authors: Stephan Steuber Ahmed AbdelRady PeterHenning Clausen
Publish Date: 2005/07/06
Volume: 97, Issue: 3, Pages: 247-254
Abstract
A polymerase chain reaction with the restriction fragment length polymorphism PCRRFLP method using universal primers complementary to the conserved region of the cytochrome b gene cyt b of the mitochondrion DNA mtDNA of vertebrates was applied to the identification of the origin of blood meals in tsetse flies Blood samples from ten potential tsetse hosts of the family bovidae cattle water buffalo red buffalo waterbuck springbok goat sheep sable antelope oryx and dikdik were included in this study Sites for appropriate restriction endonucleases cuts were chosen by pairwise alignment of the amplified 359 bp fragments A flow chart of endonucleases digestion using three restriction enzymes eg TaqI AluI and HindII for the unequivocal identification of the respective bovid species was developed A number of additional nonspecific DNA fragments attributed to the coamplification of cytochrome b pseudogenes were observed in some species eg in red buffalo and dikdik after digestion with AluI but did not hamper assignment of bovid species The detection rate of host DNA in tsetse by PCRRFLP was 100 80 60 and 40 at 24 48 72 and 96 h after in vitro feeding respectively Identification of the last blood meal was possible even when tsetse had previously fed on different hostsWe wish to thank Prof Dr Klaus Eulenberger Leipziger Zoo Dr Wolfram Rietschel ZoologischBotanischer Garten Wilhelma Stuttgart PD Dr Kai Fröhlich Institute for Zoo and Wildlife Research Berlin and Dr Andreas Ochs Zoologischer Garten Berlin for providing blood samples from different bovid species used in the study Special thanks to Mrs Angelika Wiemann for excellent technical assistance This work was funded by the Ständige Kommission für Forschung und wissenschaftlichen Nachwuchs FNK of the FU Berlin
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