Hypothesis:

Adult female wing size will decrease as phylogenetic differences increase between the variable WOWBug host (waxworm larvae) and the control WOWBug host (blowfly larvae) upon which they were reared .

Materials:

Vials with caps (10)
One bag of cotton balls
Blowfly larvae (10)
Waxworm larvae (10)
Melittobia females (75)
Ethyl acetate

Environmental Scanning Electron Microscope (ESEM)

Networked Computer with Web Browsing Software

Procedure:

1. Place five Melittobia females and two blow fly larvae in each of the first five vials.
2. Plug each vial with a cotton ball.
3. Label the vials B1 through B5.
4. Place five Melittobia females and 2 wax worm larvae in each of the next five vials.
5. Plug each vial with a cotton ball.
6. Label the vials W1 through W5.
7. Place all 10 vials in a dark area with no direct sun exposure, no significant temperature variation and minimal likelihood of jostling.
8. Allow cultures to produce mature adults for a two week period. This ensures that all adults will be from the same generation.
9. After the two week period, anesthetize the living specimens by replacing the cotton plug with a new cotton ball dipped in ethyl acetate.
10. Put vials with ethyl acetate plugs into a large, sealed jar to prevent inhalation of ethyl acetate by humans.
11. When specimens are dead, fill the vial half full of 95% ethanol and replace cotton balls with vial caps.
12. Observe the specimens with a binocular microscope by placing the vials on their sides and perform the initial adult female count, keeping in mind the fact that females have significantly larger wings than males.
13. Record the number of adult females on the vials' label and enter that data into a table for analysis. Select a representative vial from each host species (one which adheres the most closely to the normal population per vial for that host).
14. Assuming approval of the proposal, mail the two representative vials to Bugscope staff for ESEM measurement of wing and body length of females. Retain the remaining vials.
15. Using the ESEM, measure wing length and body length of each female. Enter the data into a table.
16. Perform the following calculations on females from a representative vial which has emerged from each of the two host species.

Compute the average female wing length (sum of all wing lengths divided by number of measured specimens). Record your data.

Compute the average female body length (sum of all body lengths divided by number of measured specimens). Record your data.

Compute the wing-to-body length ratio for each female. Record your data.

Compute the average wing-to-body length ratio of each vial by adding the individual ratios for each vial and dividing by the the number of specimens in each vial.

1. Compare wing length, body length and wing-to-body length ratio for each of the two cultures.
2. Analyze results. Determine whether hypothesis was supported.