Second Infection Test on D. innubila with DiNV Positive Fluid from Primary innubila Cells Day 26
Because there is some evidence that I may not be poking hard enough, I wanted to do the DiNV infection experiment again. Additionally, because sugar agar was not the right food, the mortality data was very messed up. Now I want to keep these flies on mushroom food where the control flies shouldn’t die. I also included two controls, flies that were not poked and flies poked with sterile cell culture medium. I started with 13 vials of innubila with ~10 males in each. They emerged 10/7-10/10 meaning they are 5-7 days old. I ended up making 12 experimental vials, and freezing some day 0 flies from the 13th vial.
I tried to relax as much as possible and take my time poking the flies, making sure the whole point of the needle went into the fly and a little bit more. Infections followed this general protocol however no bacteria was used, just sterile cull culture medium or the fluid from primary innubila infected with DiNV after day 26.
Infections followed as:
| Vial | Treatment | Treatment abbreviation | Replicate | Day emerged | Day Infected | time infected | fly age | Original N number |
|---|---|---|---|---|---|---|---|---|
| 1 | cell culture medium | CCM | 1 | 10/7-10/10 | 20221015 | 4:33 | 5-7 days | 10 |
| 2 | DiNV | DiNV | 1 | 10/7-10/10 | 20221015 | 4:38 | 5-7 days | 10 |
| 3 | No poke | NP | 1 | 10/7-10/10 | 20221015 | 4:45 | 5-7 days | 10 |
| 4 | DiNV | DiNV | 2 | 10/7-10/10 | 20221015 | 4:50 | 5-7 days | 10 |
| 5 | DiNV | DiNV | 3 | 10/7-10/10 | 20221015 | 4:56 | 5-7 days | 10 |
| 6 | No poke | NP | 2 | 10/7-10/10 | 20221015 | 5:01 | 5-7 days | 10 |
| 7 | DiNV | DiNV | 4 | 10/7-10/10 | 20221015 | 5:08 | 5-7 days | 10 |
| 8 | Cell culture medium | CCM | 2 | 10/7-10/10 | 20221015 | 5:14 | 5-7 days | 10 |
| 9 | No poke | NP | 3 | 10/7-10/10 | 20221015 | 5:21 | 5-7 days | 10 |
| 10 | DiNV | DiNV | 5 | 10/7-10/10 | 20221015 | 5:27 | 5-7 days | 10 |
| 11 | Cell culture medium | CCM | 3 | 10/7-10/10 | 20221015 | 5:33 | 5-7 days | 10 |
| 12 | DiNV | DiNV | 6 | 10/7-10/10 | 20221015 | 5:38 | 5-7 days | 10 |
Every 3 days flies were transferred to new food, and all dead flies were frozen individually in tubes. All transfers occurred on CO2. For information on daily mortality, see this spreadsheet. For information on the frozen flies and when they were frozen, see this spreadsheet.