STANDARD OPERATING
PROCEDURE
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version, click
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TITLE:
Mosquito Rearing Techniques
SOP NUMBER:
8.2
REVISION
NUMBER: 2
1.0 PURPOSE
The purpose of this document is to detail the
methods used in rearing Aedes aegypti.
2.0 MATERIALS USED
2.1 Cotton (nonabsorbent)
2.2 Enamel pans (9" x 14" x 2") or one-gallon
wide-mouth jars
2.3 One-gallon oyster cartons
2.4 One-half pint plastic dishes or styrofoam
cups
2.5 Nylon screen (or bobbinet cloth)
2.6 Pasteur pipettes
2.7 Pipettes for collecting pupae
2.8 Plexiglass spoons for feeding mosquito
larvae
2.9 60 mm x 15 mm petri dishes
2.10 Food for mosquito larvae (all by weight)
2.10.1 1 part rat chow - Wayne's Lab-Blox
(crushed/blended and sifted through a 40-mesh
screen)
2.10.2 1 part brewer's yeast
2.10.3 1 part lactalbumin
3.0 PROCEDURE
|
|
Day |
Amount of food(mg/container) |
Comments |
|
|
0
(Wednesday) |
100 |
Add strips of paper or cotton containing eggs to
a pan of deionized water (200-300 ml) |
|
|
1
(Thursday) |
50 |
Count 200 larvae into a petri dish and use this
dish of larvae as a reference for estimating 200
larvae for each of the remaining petri dishes.
Then transfer the larvae from each petri dish to
a container with 900 ml deionized water.
(Note: rinse petri dish thoroughly to ensure
transfer of all larvae. |
|
|
2 (Friday) |
100 |
|
|
|
3
(Saturday) |
300 |
|
|
|
4 (Sunday) |
400 |
|
|
|
5 (Monday) |
500 |
|
|
|
6
(Tuesday) |
No Food |
Pick pupae (few) |
|
|
7
(Wednesday) |
No Food |
Pick pupae (about 2/3 will have pupated) |
|
|
8
(Thursday) |
No Food |
Pick pupae (few) |
|
|
9 (Friday) |
No Food |
Pick pupae (few). Discard any remaining
larvae |
If the water becomes "cloudy" or if "scumming"
occurs, the larvae will usually die. This usually
results from over-feeding of the larvae or from not
having enough larvae in the container.
4.0 PROCEDURE FOR PRODUCTION OF EGGS
4.1 A small (about one-half pint size) plastic
dish or styrofoam cup is attached to the inside of a
one-gallon oyster carton.
4.2 The dish/cup is lined with a strip of paper
towel and approximately 800 pupae (female and male
combined) are added.
4.3 The center part of the lid is removed and
replaced with nylon screening. The lid is also taped
to the carton as a precautionary measure. Adults
emerge within 2 days.
4.4 As soon as possible after all of the adults
have emerged, the water is aspirated from the dish/cup
by making a small hole in the nylon screening and
inserting a pipette for the aspiration.
4.5 Cotton is used to plug the hole in the
screen. The mosquitoes are allowed to feed on sugar
cubes and water contained in cotton which are placed on
the nylon screen.
4.6 Three to five (preferably 4) days later, the
mosquitoes are given a "normal" blood meal.
4.7 On the following day, cotton that is
saturated with deionized water and wrung out is placed
on top of the nylon screen, and the females will lay
eggs on the cotton within 48 hours.
4.8 Two to three days later, the cotton is
removed and allowed to dry slowly. These eggs may be
stored in a cool, dry place for several months.
5.0 TEMPERATURE AND HUMIDITY OF THE INSECTARY
5.1 The temperature in the room (specifically,
near the cartons of infected mosquitoes) should be
maintained at approximately 80
± 9°F
(27 ± 5°C)
and the humidity should be maintained at approximately
80 ± 15% (R.H.).
5.2 A daily record of the temperature and
humidity will be recorded on the Temperature/Humidity
log and will be kept on file.
|