Urbanization Spectrum of Vector Culex Mosquitos Research Opportunity

 During the last week of June, my mentor Ben Gregory was out of the office for a vacation. As such, I have been tasked with catching up on the backlog of PCR and gel electrophoresis necessary to identify the difference between Cx .  restuans and Cx. pipens . Normally, during the week, I have performed two PCR’s per week; however, with six different trap nights coming in a week, and the preference to run smaller, 36 sample groups from each trap night, it has been challenging to keep up.  On Monday, Izi and I ran three PCR’s. Then I had to go set up traps at the last three sites of June; Hillcrest Park, SERC, and Church of the Holy City.  The Hillcrest Park gravid trap was set up between two trees next to a pond. The park happened to be windy, and I got dust in my eyes! On Tuesday, we ran more PCRs and got 100% amplification when viewing the gels. We also had to make more gels for gel electrophoresis on Wednesday. Here is the 100% amplification gel! I was so proud of the teamwork Izi and

  After nearly a month working at the Department of Entomology, I have got the hang of all the molecular work done to assist Ben with his research, including the most technical, polymerase chain reaction, otherwise known as PCR. PCR is a molecular technique used to amplify specific segments of DNA in vitro. Thermal cycling separates dsDNA into ssDNA and primers are used to select the DNA segment that will be amplified. In this case, multiple primers used to segment multiple target sequences to identify the different species within the Culex genus. The Primers are annealed to the template ssDNA to allow for the Taq, a DNA polymerase, to add onto the primers dNTPs to synthesize the segment of interest. This process of denaturation, annihilation and elongation repeats until billions of segments are replicated. Having a thorough understanding of the process of PCR provides a framework for how to set up a PCR in practice, as well as a great appreciation. The lab work of PCR is simple, unlik

During the third week, Izi and I learned about how to prepare a “squish prep” otherwise known as a DNA extraction. This step was fun to learn because it involves more technical squish prep where we must dissect the mosquitoes by separating the thorax from their abdomen.  Picture of dissected Culex mosquito. Abdomen is used for squish prep and kept in a strip tube while the thorax is kept in a separate tube as the individual. To prepare for this first part of DNA extraction, we set up an ice bucket to keep the thoraxes, and four strip tubes to keep the abdomen. The tubes for the thoraxes are much bigger and are labeled with the mosquito's names, which include the year, their trap night, and the number of the individual. We organize the individual abdomens using a drawn diagram. Shown below. This is an example of the notebook setup and the microscope used to dissect the mosquitos. the beaker contains a 10% bleach solution for disinfecting the Petri dish and forceps. Generally, it is

  After catching mosquitoes, the next step in the protocol is to sort mosquitos underneath a microscope. So to start off the second week of the internship, Ben showed me the proper way to identify different insects in the trap as well as the key field features of Culex mosquitoes! The first thing to look for to identify mosquitoes from other invertebrates are their scaly wings. They have hair-like appendages on the very ends of their wings. They also have a long proboscis, a long tubular mouthpiece used to suck up the blood of other animals. The insect above lacks scaly wings while the insect below is a Culex mosquito.  Culex in particular focuses on avians and people, though different subspecies have a preference for each group with Culex pipiens and Culex quinquefasciatus biting both humans and birds and people. The other species in the genus, Culex r estuans tends to feed primarily on birds. After identifying the insect to be a mosquito, the next field characteristic of Culex I w