Protocol for Mosquito Rearing (A. gambiae)

Summary

This video illustrates the general techniques used to rear Anopheles gambiae in the laboratory. The methods for caring for laboratory mosquitoes are demonstrated through all stages of the organism's life cycle from larvae to pupae to blood-feeding adults.

Abstract

This protocol describes mosquito rearing in the insectary. The insectary rooms are maintained at 28°C and ~80% humidity, with a 12 hr. day/night cycle. For this procedure, you'll need mosquito cages, 10% sterile sucrose solution, paper towels, beaker, whatman filter paper, glass feeders, human blood and serum, water bath, parafilm, distilled water, clean plastic trays, mosquito food (described below), mosquito net to cover the trays, vacuum, and a collection chamber to collect adults.

Protocol

Mosquito food:

1. Food A: grounded fish food (Aquaricare). A small pinch needs to be added.
2. Food B: grounded CAT food (Purina). A small pinch needs to be added.
3. Food C: cat food (Purina). Two tablets needs to be added.

Blood:

Animal or human blood can be used to rear mosquitoes. Fresh blood is collected with a syringe and put in a sterile 15 ml Falcon tube containing 1ml of CPD (anticoagulant) for each 10 ml of blood, then mixed gently and centrifuged at room temperature at 2000 rpm for 5 min. The supernatant is discarded, taking care to remove the buffy coat which comprises other ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Food A		Aquaricare		Grounded fish food. A small pinch needs to be added.
Food B		Purina		Grounded cat food. A small pinch needs to be added.
Food C		Purina		Cat food. Two tablets needs to be added
RBC				prepared red blood cells: 40% hematocrit in human serum
RPMI	Medium
CPD				anticoagulant
serum				O+ human serum
Mosquito feeder
parafilm
Anopheles gambiae				mosquitos
10% sucrose				mosquito food
insectary