Biology: A Molecular Approach, Laboratory

1. Biology: A Molecular Approach, Laboratory Lab 2

2. Today’s Plan • Attendance • Handout • Introduction to Your Research Project • Webquest on Mosquito Biology • Mosquito Trapping • Mosquito Identification

3. Introduction to your Research Project

4. Introduction to your Research Project • Questions • In our region: • Which mosquito species are infected with the bacteria Wolbacchia pipientis? • Which mosquitoes species are infected with the heartworm Dirofilaria immitis? • Methods • Trap mosquitoes • Identify sex and genus/species of trapped mosquitoes • Use PCR to detect presence of Wolbacchia pipientis and/or Dirofilaria immitis in identified mosquitoes

5. Webquest on Mosquito Biology • Assignment • Goal is to help you learn about • Mosquito biology and ecology • The nematode Dirofilaria immitis • The bacteria Wolbacchia pipientis • The WO phage • You will use this information to prepare your scientific writeups on your research project • You also will be tested on this during a lecture exam

6. Webquest on Mosquito Biology • Complete this assignment individually! • After lab, download the file “Mosquito Biology and Ecology” • Angel; JCC Biotech; BAMA: A Molecular Approach, Laboratory; Lab 2 • Follow instructions exactly as stated in the file • Type your answers directly into the file and submit your file into the Angel drop box called “Mosquito Biology and Ecology Drop Box”: • Angel; JCC Biotech; BAMA: A Molecular Approach, Laboratory; Lab 2 • Due date is Wednesday,September 12, 2:40pm

7. Mosquito Trapping • In a couple of weeks, you will need to writeup how to trap mosquitos for your scientific writeup • Two places to obtain this information • File called “Mosquito Trapping” • Angel, JCC Biotech, BAMA: A Molecular Approach, Laboratory, Lab 2 • Jamie Haight • Take notes on his presentation today

8. Mosquito Trapping • Jamie Haight’s Presentation • Walk to the woods to set the traps

9. Mosquito Morphology • Two Resources • Identification Guide to the Mosquitoes of Connecticut • Is in the ring binder at your lab benches • p. 37 - 39 • File called “Mosquito Identification” • Angel, JCC Biotech, BAMA: A Molecular Approach, Laboratory, Lab 2 • Get your laptop on and logon to the Internet • Find and open this file now • p. 1 – 5 • Don’t memorize the morphology; just refer to these two resources when you come across terms as we do mosquito sexing and genus/species identification

10. Mosquito Sexing • Do this activity in your group • Open the file called “Mosquito Identification” • Angel, JCC Biotech, BAMA: A Molecular Approach, Laboratory, Lab 2 • Read the section on “Mosquito Sexing” on p. 1 • Identify the sex of the mosquitoes in your practice tube • Use the dissecting microscope • Gently empty the tube of mosquitoes into the petri dish • Use forceps to gently manipulate and view the mosquitoes • We’ll help you use the scopes • Keep females in the petri dish • Put males back into the practice tube

11. Mosquito Genus and Species Identification • You need to learn how to identify the genus and species of the most common local mosquitoes • Anopheles genera • Anopheles quadrimaculatus • Anopheles punctipennis • Anopheles walkeri • Culiseta genera • Culiseta melanura • Culiseta morsitans • Culex genera • Culex territans • Culex pipiens • Culex restuans • Coquillettidia genera • Coquillettidia perturbans

12. Mosquito Genus and Species Identification • Two Resources • Identification Guide to the Mosquitoes of Connecticut • Is in the ring binder at your lab benches • File called “Mosquito Identification” • Angel, JCC Biotech, BAMA: A Molecular Approach, Laboratory, Lab 2 • Should have this file already open • Don’t memorize these resources, just learn how to use them to identify the mosquito genus and species

13. Mosquito Genus and Species Identification • Do this activity in your group • Identify the Anopheles genera • Do step 1 on p. 40 of the Identification Guide to the Mosquitoes of Connecticut • You do not need to look at the scutellum • Identify the Anopheles species • Use the information on p. 7 of the filed called “Mosquito Sexing and Identification”

14. Mosquito Genus and Species Identification • Do this activity in your group • Identify Coquillettidia perturbans • Do step 8 on p. 43 of the Identification Guide to the Mosquitoes of Connecticut • See also the information on p. 10 – 11 of the filed called “Mosquito Sexing and Identification”

15. Mosquito Genus and Species Identification • Do this activity in your group • Identify the Culiseta and Culex genera • Do step 9 on p. 43 of the Identification Guide to the Mosquitoes of Connecticut

16. Mosquito Genus and Species Identification • Do this activity in your group • Identify the Culex species • Do steps 1 and 4 on p. 57 – 58 of the Identification Guide to the Mosquitoes of Connecticut • See also the information on p. 9 of the filed called “Mosquito Sexing and Identification”

17. Mosquito Genus and Species Identification • Do this activity in your group • Identify the Culiseta species • Do steps 1 on p. 59 of the Identification Guide to the Mosquitoes of Connecticut • See also the information on p. 8 of the filed called “Mosquito Sexing and Identification”

18. Supplies • Mosquito trapping • CO2 tank • Dry ice machine • 6 dry ice cubes • Cooler to hold dry ice at -80C until used • 2 CO2 traps • Batteries • Battery charger • Sheets to record when set and what was collected • Mosquito Trapping File • Mosquito ID • 6 Dissecting microscopes • 6 Forceps • 6 Petri dishes • Mosquitoes sorted into 6 plastic tubes in a rack which are a mix of unknowns • 6 Printouts of Identification Guide to the Mosquitoes of Connectictut • Mosquito Sexing and Identification File • General • Laptops • Mosquito Biology and Ecology Webquest File