Second Infection of D. innubila with 16Cq DiNV For Freezing for Viral Load on Day 0, Day 3, Day 5, and Day 10

This the second replicare experiment to 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 planned to use:

  • 10 flies infected with DiNV for day 0, 6 sterile poke flies for day 0
  • 20 flies infected with DiNV for day 3, 6 sterile poke flies for day 3
  • 20 flies infected with DiNV for day 5, 6 sterile poke flies for day 5
  • 20 flies infected with DiNV for day 10, 6 sterile poke flies for day 10

Flies were separated out into vials of ~7 or ~11 individuals the day before on the 5thnd 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
A innubila sterile poke SP male 5-30/6-1 20230606 3:14 6-7 days 5
B innubila sterile poke SP male 5-30/6-1 20230606 3:18 6-7 days 6
C innubila sterile poke SP male 5-30/6-1 20230606 3:22 6-7 days 6
D innubila sterile poke SP male 5-30/6-1 20230606 3:26 6-7 days 6
E innubila sterile poke SP female 5-30/6-1 20230606 3:30 6-7 days 6
F innubila sterile poke SP female 5-30/6-1 20230606 3:34 6-7 days 6
G innubila sterile poke SP female 5-30/6-1 20230606 3:38 6-7 days 6
H innubila sterile poke SP female 5-30/6-1 20230606 3:42 6-7 days 6
I innubila 16 Cq DiNV 16DiNV male 5-30/6-1 20230606 3:46 6-7 days 9
J innubila 16 Cq DiNV 16DiNV male 5-30/6-1 20230606 3:51 6-7 days 10
K innubila 16 Cq DiNV 16DiNV male 5-30/6-1 20230606 3:57 6-7 days 10
L innubila 16 Cq DiNV 16DiNV male 5-30/6-1 20230606 4:03 6-7 days 10
M innubila 16 Cq DiNV 16DiNV male 5-30/6-1 20230606 4:07 6-7 days 9
N innubila 16 Cq DiNV 16DiNV male 5-30/6-1 20230606 4:12 6-7 days 10
O innubila 16 Cq DiNV 16DiNV male 5-30/6-1 20230606 4:17 6-7 days 10
P innubila 16 Cq DiNV 16DiNV female 5-30/6-1 20230606 4:23 6-7 days 10
Q innubila 16 Cq DiNV 16DiNV female 5-30/6-1 20230606 4:27 6-7 days 10
R innubila 16 Cq DiNV 16DiNV female 5-30/6-1 20230606 4:32 6-7 days 10
S innubila 16 Cq DiNV 16DiNV female 5-30/6-1 20230606 4:37 6-7 days 10
T innubila 16 Cq DiNV 16DiNV female 5-30/6-1 20230606 4:41 6-7 days 10
U innubila 16 Cq DiNV 16DiNV female 5-30/6-1 20230606 4:45 6-7 days 10
V innubila 16 Cq DiNV 16DiNV female 5-30/6-1 20230606 4:50 6-7 days 10

For all flies to be frozen on day 0, they were put into individual tubes for freezing.

sample_ID vial_from sex treatment day_frozen alive_when_frozen?
1 A male SP day0 yes
2 A male SP day0 yes
3 A male SP day0 yes
4 A male SP day0 yes
5 A male SP day0 yes
6 E female SP day0 yes
7 E female SP day0 yes
8 E female SP day0 yes
9 E female SP day0 yes
10 E female SP day0 yes
11 E female SP day0 yes
12 I male 16CqDiNV day0 yes
13 I male 16CqDiNV day0 yes
14 I male 16CqDiNV day0 yes
15 I male 16CqDiNV day0 yes
16 I male 16CqDiNV day0 yes
17 I male 16CqDiNV day0 yes
18 I male 16CqDiNV day0 yes
19 I male 16CqDiNV day0 yes
20 I male 16CqDiNV day0 yes
21 P female 16CqDiNV day0 yes
22 P female 16CqDiNV day0 yes
23 P female 16CqDiNV day0 yes
24 P female 16CqDiNV day0 yes
25 P female 16CqDiNV day0 yes
26 P female 16CqDiNV day0 yes
27 P female 16CqDiNV day0 yes
28 P female 16CqDiNV day0 yes
29 P female 16CqDiNV day0 yes
30 P female 16CqDiNV day0 yes

All information on the infections can be found here All information about the freezing of the flies can be found here