1. Biology: A Molecular Approach, Laboratory

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
Mosquito
Heartworm nematode (Dirofilaria immitis)
(Wolbacchia pipientis)
Bacteria
WO Phage

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”:
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
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