University of Wisconsin - Green Bay


Research Council

Research
Success

Uwe Pott

Spring 2011

"How Much Heartworm Prophylaxis Is Enough?"

Grant in Aid of Research

Final Report: "Fund use: The funds were used to purchase materials for DNA purification from mosquitoes (DNA extraction kits) and for running polymerase chain reactions on the purified DNAs (Taq polymerase and PCR reaction tubes), as specified in the grant application. Project outcome: The goal of the work supported by this grant was to overcome a road block in my research project investigating the prevalence of infective heartworm larvae in mosquitoes in Northeast Wisconsin. In order to perform a reliable, large-scale survey of the occurrence of the parasite during the mosquito season, we had to establish a method that could detect a single heartworm larva in a pool of up to 100 mosquitoes. Brad Beaumier, a highly talented undergraduate student who has moved on to veterinary school, and I were able to achieve the necessary sensitivity of the assay during our collaboration in the fall of 2011 and the spring of 2012. Brad presented our work at the 12th Annual UW System Symposium for Undergraduate Research and Creative Activity on April 27, 2012 at UW-Parkside. The following abstract of his talk was printed in the symposium program. Abstract: Heartworm Detection in Wild-Caught Mosquitoes from Northeastern Wisconsin Heartworm disease in dogs and wild canids is caused by the nematode Dirofilaria immitis. This parasitic roundworm must progress through several larval stages in a mosquito host before the infective larva (L3) can be transmitted to the definitive host. In the dog the L3 develops into the adult worm, which settles in major blood vessels near the heart and may ultimately cause heart failure. The goal of this study is to estimate the risk of heartworm infection for dogs in Northeastern Wisconsin during the mosquito season by determining the prevalence of heartworm larvae in the mosquito hosts. Our experimental protocol includes trapping the insects in light traps with carbon dioxide as additional attractant, preparing crude lysates from pools of mosquitoes, and detecting heartworm DNA in these lysates using the polymerase chain reaction (PCR). Currently we are testing if our protocol will allow us to detect reliably the DNA of a single L3, the smallest infective unit, in a large pool of mosquitoes. Thus far, we have been able to amplify heartworm-specific DNA fragments out of pools of 100 mosquitoes spiked with 6 ng of heartworm DNA (courtesy of Dr. Kirsten Crossgrove, UW-Whitewater) or one L3 (courtesy of NIAID/NIH Filariasis Research Reagent Resource Center, University of Georgia). We therefore have established the sensitivity necessary to test a large number of mosquitoes caught during the summer of 2011 in Green Bay, WI, for the presence of larval stages of the heartworm parasite."


Spring 2007

"Identification of the rKr2 Nuclear Localization Signal in the Molecular Biology Lab Course"

Grant for Integrating Research and Teaching


Fall 2006

"The Risk of Dogs in our area to be infected with the canine heartworm Dirofilaria immitis"

Grant in Aid of Research


Fall 2004

"Risk Assessment of Canine Heartworm Infection in NE Wisconsin"

Grant in Aid of Research


Spring 2003

"Large-scale Expression of rKr2 Fusion Proteins in E. coli"

Grant in Aid of Research


Spring 2002

"Identification of a Nuclear Localization Signal in the Zinc Finger Protein rKr2"

Grant for Integrating Research and Teaching


Spring 2002

"Affinity Purification of a DNA Target Site for rKr2"

Grant in Aid of Research


Spring 2001

"Identification and Characterization of a DNA Binding Site for the Zinc Finger Protein rKr2"

Grant in Aid of Research


Spring 2000

"Identification of a Nuclear Localization signal in Zinc Finger Protein rKr2"

Grant in Aid of Research