Edmonds, Sarah L.  (2005). Proteomic analysis of the effect of parasitism by Uvulifer ambloplitis on bluegill (Lepomis macrochirus) plasma proteins. MS thesis, University of Akron, Ohio  (Londraville)

Abstract:

 Bluegill sunfish, Lepomis macrochirus are commonly parasitized by the black grub, Uvulifer ambloplitis, which is known to cause black-spot disease in Centrarchid fishes.   To date no studies have evaluated the physiological effect of U. ambloplitis on bluegill.  The aim of this study was to determine the effects of parasitism by U. ambloplitis on protein expression in bluegill plasma using proteomics.  Two experimental groups were compared, bluegill sunfish without external parasites and bluegill experimentally infected with U. ambloplitis.  Blood was collected at different time intervals, and plasma prepared.  Albumin and immunoglobins were removed, and the remaining protein was extracted using a commercial kit.  Proteins were separated by isoelectric point and then by molecular weight using two dimensional gel electrophoresis.  With care, 2-DE gels are highly reproducible: pixel volume for any given spot changes only 12-13% among repeat gels of the same sample.  A total of 544 bands were detected using 2-DE analysis software.  Different analyses were conducted and there were a total of 50 spots that were considered proteins of interest.  Those proteins and several landmark proteins were identified using mass spectrometry.  The acute phase response is part of the initial immune response to infection.  Several proteins were identified that can be related to previously described acute phase proteins, including complement component C3, transferrin, and apolipoprotein.  However, many proteins of interest that were sequenced could not be identified due to the lack of homology of bluegill protein sequences and those of other vertebrates. 

Full Thesis on file at UA.