Here is the link to the worksheet saved in aggiemail drive:
https://docs.google.com/spreadsheets/d/1R7ybhHUjTD8pSqL662YFq3ohaLCtUY-FiNO2Ps1DxJM/edit#gid=0
On July 23rd 2017
I cleared all the mating cages from the chamber. The bugs were all dead except one female from BST which had crooked wings so I froze it.
On July 21st 2017
The bugs are all almost dead. I have frozen the dead ones in -80. Today I just took a few eggs and tried to measure their size. I see that the ones from SKI are average 0.722 and ones from BST are 0.65. So there is a difference in size. I hope the BST ones hatch. Also, both these are from lupine, long day treatment. I did not find any in SD. aaahhhh this experiment might finally end. I think I will record the adult sizes and maybe also record the crooked winged etc data.
On July 17th 2017
Interestingly the buddleia bush was very weird. A bunch of bugs died. I did find a bunch of eggs in the SKI-L-L treatment. I have a feeling the bugs reared on alflafla wont do anything. All the BST-M-L bugs died today.
On July 13th 2017
I put the buddelia bush in the cages today. I put one pot per cage. The butterflies seem more active around it. I also found eggs in SKI-L-L and SKI-L-S. I hope they all mate and lay lots of eggs.
On July 12th 2017
Here are somethings to keep note for this experiment:
I used gatorad as nectar source
I provided both alfalfa and lupine in the mating cages and dabbed some gatorad on the cages
Few bugs came out all wrinkly
I see a difference in size in adults reared on lupine(bigger) and alfalfa(smaller)
A bunch of bugs have died. These just stop developing and turn dark and I just find them lying dead the next day.
Before transferring the bugs to the mating cages they were kept in small round boxes. There eclosion date was recorded and then they were transferred to the mating cages.
For bugs that died, I did not record the date I found them dead. I just recorded that they died. So this might be treated as survival till pupation(?).
NOTES ABOUT MATING CAGES:
All bugs seem to be mobile and alive
I set up cages with two conical flasks with alfalfa and lupine plants
Matt suggested that the plants should be raised up and touching the top of the cage so that they are accessible to the females. So I placed the conical flasks on a small thermacol box to elevate the plants.
The temperature of the walk in chamber is 80F and I cannot figure out what is the humidity. The light conditions there are 11H dark and 13 H light. Light comes on at 6am and goes off at 7 pm.
I provided gatorad in a small tube but bugs dont seem to access it so I smear gatorad on the cage itself and spray them with water. I spray the cage with water thrice a day.
On July 9th 2017
I found this old paper which described mating cages for Karner blues. Here are the important details:
Housing of butterflies: In the laboratory, butterflies were transferred to
aluminum frame cages (61 x 61 x 61 cm) with 32 mesh Lumite screen (Bio-
Quip Products, Gardena, CA). We opened each envelope inside the cage and
allowed the female to walk out onto lupine foliage (described below). Butter-
flies were caged together by site. Cages were kept on fluorescent-lighted
shelves in a walk-in environmental chamber maintained a t 24"-26"C, with an
18:6 hr 1ight:dark photoperiod, and relative humidity of 57-68 percent.
We provisioned each cage with a water source, partial shading, nectar source, and ovipositional site. The water source was a wet sponge cut to
tightly fit the bottom of a petri dish (100 x 15 mm). One sponge was provided
per cage, and was moistened daily. Any standing water or condensation was
wiped up immediately, to prevent butterflies from becoming trapped or
drowning (Lane and Welch 1994). We provided partial shading by placing lay-
ers of paper towels over one corner of the top of the cage.
The nectar source was a 5 percent honey: 95 percent water solution (Lane
and Welch 1994). The solution was placed in a sterile 150-ml flask, and then
sealed with parafilm. Cotton dental wicking (Accu Bite Dental Supply Inc.,
East Lansing, MI) was pushed partially into the flask through the parafilm,
leaving 3-5 cm of wicking protruding, to provide a suitable place for butter-
flies to perch and feed. We provided two nectar flasks in each cage, and re-
placed them every 2 days.
The ovipositional site consisted of a wild lupine stem, 20-30 cm tall, with
flowers and leaves, in a water-filled 250-ml flask with a parafilm seal. We
placed two flasks with lupine in each cage, and replaced them every 2 days
with fresh lupine.
We housed the females for 5 days in the cages, and then returned all sur-
vivors to their original collection sites. Female butterflies were transported in
a ca. 20°C cooler, in glassine envelopes and plastic containers as previously
described, to the appropriate site. At the sites, we released each female by
opening the envelope near a lupine plant, and allowing the butterfly to walk
onto a leaf.
I might use these methods to maximize mating success. Lets see if it works!! Right now they are not mating. :(
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--- Well my experiment got messed up and now I have only few samples left.
Here are some things I have noticed so far:
The larvae on lupine are definitely developing way faster and are already going into pupation. The LD ones are mostly into pupation. The SD ones are developing a little slowly. The alfalfa ones in both treatments are developing slowly.
--I need to remember since the larvae are developing at a different rate on lupine and alfalfa, there might be weird differences in RNA expression and I need to make sure I account for these differences if I get RNA data.
--There are some bugs which were misplaced with the plants. I am still letting them grow and will set up cages for them accordingly.