Infection D. innubila with 16Cq DiNV For Freezing for Viral Load on Day 0, Day 3, Day 5, and Day 10
This experiment is to sort of get at what percentage of flies are infected at what stage during the infection. The plan is to freeze infected flies at day 0, day 3, day 5, and day 10 after being poked. I can then do PCR to test how many were infected, and do qPCR down the line if results look interesting to look at viral load. For this I am using males and females to see their potential differences.
For males and females I used:
- 10 flies infected with DiNV for day 0, 5 sterile poke flies for day 0
- 20 flies infected with DiNV for day 3, 5 sterile poke flies for day 3
- 20 flies infected with DiNV for day 5, 5 sterile poke flies for day 5
- 20 flies infected with DiNV for day 10, 5 sterile poke flies for day 10
Flies were separated out into vials of ~6 or ~11 individuals two days before on the 22nd so they would be easier to sex and count during infections.
All sterile poke flies were done first to avoid any possible contamination issues. The 16Cq fluid was thawed on ice and kept on ice for the duration of the infections.
Infection specifics below:
| vial | Species | Treatment | Treatment_abreave | sex | Day_emerged | Day_Infected | time_infected | fly_age | Original_N_number | day_to_be_frozen |
|---|---|---|---|---|---|---|---|---|---|---|
| A | innubila | sterile poke | SP | male | 5/17-5/18 | 20230524 | 12:20 | 6-7 days | 5 | day0 |
| B | innubila | sterile poke | SP | male | 5/17-5/18 | 20230524 | 12:26 | 6-7 days | 5 | day3 |
| C | innubila | sterile poke | SP | male | 5/17-5/18 | 20230524 | 12:30 | 6-7 days | 5 | day5 |
| D | innubila | sterile poke | SP | male | 5/17-5/18 | 20230524 | 12:33 | 6-7 days | 5 | day10 |
| E | innubila | sterile poke | SP | female | 5/17-5/18 | 20230524 | 12:37 | 6-7 days | 5 | day0 |
| F | innubila | sterile poke | SP | female | 5/17-5/18 | 20230524 | 12:40 | 6-7 days | 5 | day3 |
| G | innubila | sterile poke | SP | female | 5/17-5/18 | 20230524 | 12:43 | 6-7 days | 5 | day5 |
| H | innubila | sterile poke | SP | female | 5/17-5/18 | 20230524 | 12:46 | 6-7 days | 5 | day10 |
| I | innubila | 16 Cq DiNV | 16DiNV | male | 5/17-5/18 | 20230524 | 12:50 | 6-7 days | 9 | day0 |
| J | innubila | 16 Cq DiNV | 16DiNV | male | 5/17-5/18 | 20230524 | 12:56 | 6-7 days | 10 | day3 |
| K | innubila | 16 Cq DiNV | 16DiNV | male | 5/17-5/18 | 20230524 | 1:01 | 6-7 days | 10 | day5 |
| L | innubila | 16 Cq DiNV | 16DiNV | male | 5/17-5/18 | 20230524 | 1:08 | 6-7 days | 10 | day10 |
| M | innubila | 16 Cq DiNV | 16DiNV | male | 5/17-5/18 | 20230524 | 1:14 | 6-7 days | 10 | day3 |
| N | innubila | 16 Cq DiNV | 16DiNV | male | 5/17-5/18 | 20230524 | 1:20 | 6-7 days | 10 | day5 |
| O | innubila | 16 Cq DiNV | 16DiNV | male | 5/17-5/18 | 20230524 | 1:25 | 6-7 days | 10 | day10 |
| P | innubila | 16 Cq DiNV | 16DiNV | female | 5/17-5/18 | 20230524 | 1:32 | 6-7 days | 10 | day0 |
| Q | innubila | 16 Cq DiNV | 16DiNV | female | 5/17-5/18 | 20230524 | 1:37 | 6-7 days | 10 | day3 |
| R | innubila | 16 Cq DiNV | 16DiNV | female | 5/17-5/18 | 20230524 | 1:41 | 6-7 days | 10 | day5 |
| S | innubila | 16 Cq DiNV | 16DiNV | female | 5/17-5/18 | 20230524 | 1:46 | 6-7 days | 10 | day10 |
| T | innubila | 16 Cq DiNV | 16DiNV | female | 5/17-5/18 | 20230524 | 1:51 | 6-7 days | 10 | day3 |
| U | innubila | 16 Cq DiNV | 16DiNV | female | 5/17-5/18 | 20230524 | 1:56 | 6-7 days | 10 | day5 |
| V | innubila | 16 Cq DiNV | 16DiNV | female | 5/17-5/18 | 20230524 | 2:00 | 6-7 days | 10 | day10 |
For all flies to be frozen on day 0, they were put into individual tubes for freezing. 1 male fly from vial I flew off the CO2 pad before freezing, so that one set of flies will have 1 less replicate.
| sample_ID | vial_from | sex | treatment | day_frozen | alive_when_frozen? |
|---|---|---|---|---|---|
| 1 | A | male | sterile poke | day0 | yes |
| 2 | A | male | sterile poke | day0 | yes |
| 3 | A | male | sterile poke | day0 | yes |
| 4 | A | male | sterile poke | day0 | yes |
| 5 | A | male | sterile poke | day0 | yes |
| 6 | E | female | sterile poke | day0 | yes |
| 7 | E | female | sterile poke | day0 | yes |
| 8 | E | female | sterile poke | day0 | yes |
| 9 | E | female | sterile poke | day0 | yes |
| 10 | E | female | sterile poke | day0 | yes |
| 11 | I | male | 16Cq DiNV | day0 | yes |
| 12 | I | male | 16Cq DiNV | day0 | yes |
| 13 | I | male | 16Cq DiNV | day0 | yes |
| 14 | I | male | 16Cq DiNV | day0 | yes |
| 15 | I | male | 16Cq DiNV | day0 | yes |
| 16 | I | male | 16Cq DiNV | day0 | yes |
| 17 | I | male | 16Cq DiNV | day0 | yes |
| 18 | I | male | 16Cq DiNV | day0 | yes |
| 19 | I | male | 16Cq DiNV | day0 | yes |
| 20 | NA | NA | NA | NA | NA |
| 21 | P | female | 16Cq DiNV | day0 | yes |
| 22 | P | female | 16Cq DiNV | day0 | yes |
| 23 | P | female | 16Cq DiNV | day0 | yes |
| 24 | P | female | 16Cq DiNV | day0 | yes |
| 25 | P | female | 16Cq DiNV | day0 | yes |
| 26 | P | female | 16Cq DiNV | day0 | yes |
| 27 | P | female | 16Cq DiNV | day0 | yes |
| 28 | P | female | 16Cq DiNV | day0 | yes |
| 29 | P | female | 16Cq DiNV | day0 | yes |
| 30 | P | female | 16Cq DiNV | day0 | yes |
All information on the infections can be found here All information about the freezing of the flies can be found here