Biology: A Biology: A Molecular Molecular Approach, Approach, LaboratoryLaboratory

Biology: A Biology: A Molecular Molecular Approach, Approach, LaboratoryLaboratory

Lab 2Lab 2

Today’s PlanToday’s PlanToday’s PlanToday’s Plan

• Attendance• Handout• Introduction to Your Research Project• Webquest on Mosquito Biology• Mosquito Trapping• Mosquito Identification

Introduction to your Research ProjectIntroduction to your Research ProjectIntroduction to your Research ProjectIntroduction to your Research Project

Introduction to your Research ProjectIntroduction to your Research ProjectIntroduction to your Research ProjectIntroduction 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

Webquest on Mosquito BiologyWebquest on Mosquito BiologyWebquest on Mosquito BiologyWebquest 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

Webquest on Mosquito BiologyWebquest on Mosquito BiologyWebquest on Mosquito BiologyWebquest 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

Mosquito TrappingMosquito TrappingMosquito TrappingMosquito 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

Mosquito TrappingMosquito TrappingMosquito TrappingMosquito Trapping

• Jamie Haight’s Presentation

• Walk to the woods to set the traps

Mosquito MorphologyMosquito MorphologyMosquito MorphologyMosquito 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

Mosquito SexingMosquito SexingMosquito SexingMosquito 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

Mosquito Genus and Species IdentificationMosquito Genus and Species IdentificationMosquito Genus and Species IdentificationMosquito 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

Mosquito Genus and Species IdentificationMosquito Genus and Species IdentificationMosquito Genus and Species IdentificationMosquito 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

Mosquito Genus and Species Mosquito Genus and Species IdentificationIdentificationMosquito Genus and Species Mosquito Genus and Species IdentificationIdentification

• 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”

Mosquito Genus and Species Mosquito Genus and Species IdentificationIdentificationMosquito Genus and Species Mosquito Genus and Species IdentificationIdentification

• 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”

Mosquito Genus and Species Mosquito Genus and Species IdentificationIdentificationMosquito Genus and Species Mosquito Genus and Species IdentificationIdentification

• 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

Mosquito Genus and Species Mosquito Genus and Species IdentificationIdentificationMosquito Genus and Species Mosquito Genus and Species IdentificationIdentification• 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”

Mosquito Genus and Species Mosquito Genus and Species IdentificationIdentificationMosquito Genus and Species Mosquito Genus and Species IdentificationIdentification• 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”

SuppliesSuppliesSuppliesSupplies

• 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