Went in to UW to continue imaging Laura’s slides. Imaged slides 3-10. Here are the samples that I either couldn’t find gonad tissue in or I wasn’t quite sure about:
Oly 3 slide: HL-6_13 (not sure if I captured gonad tissue), HL-6_14 (none, no picture)
Oly 6 slide: NF-6_17 (none, no picture), NF-6_19 (not sure if I captured gonad tissue)
Oly 7 slide: NF-6_18 (not sure if I captured gonad tissue)
Oly 8 slide: SN-6_18 (not sure if I captured gonad tissue)
I will go through all of my noted questionable slides when I’m done.
Grace showed me how to image histology slides today in the same way as she did the post-overwintering samples. I will image all of the slides at low (4x) and high (40x) magnification with the high magnification looking at the gonads of the oyster and the low magnification being the zoomed out image of the same place. Some of the oysters were split between slides or moved around after Laura sent them in, but between her histology key and her images of what the oysters looked like in the cassettes I’ve been able to figure out which oysters are which.
The pictures are labeled in the same way as Laura’s histology key (location-overwintering temp-sample #) and then labeled as either high or low (magnification). I was unable to find any gonad tissue in a couple of the smaller oysters and I will have to go back and figure out a way to make a note of that. The two I was unable to find gonad tissue in thus far were HL-10_14 and HL-6_11.
Here is the URL for the drive where I will be uploading all my images:
Went into UW today and checked all the silos for larvae. Two out of the 13 individual oysters had spawned with one having thousands of larvae and one having only a few. Also possible presence ciliates, but there were larvae there!
Here are my counts:
In addition, I talked with Steven and Grace about how to image all of Laura’s histology slides from after the OA treatment. From there I will work to go through them all and look at gonad maturity between treatments and after just the overwintering as well as after the OA treatments.
Went in the check out the set-up at UW. Didn’t see any larvae, but I’m going to stop back in tomorrow and check again! It’s a little hard to tell if there are larvae in the silos as my flashlight doesn’t shine through the sides, but I looked at the water in a few of them under the microscope and couldn’t see anything.
I’ll have to figure out what to do if one of the groups that isn’t divided up into separate containers has larvae in it and how long to keep keep the oysters where they are before switching which group is split up into individual silos.
Today I went checked out the water my oysters were put in yesterday and the K-10 pH 8 water had a ton of larvae in it! The oysters had been moved to smaller containers of water early this morning, so I was able to sample, count, and image the larvae using yesterdays water.
Some images of the new larvae:
Moving forward we were thinking about designing an experiment where we observe individual oysters as then spawn. This would allow us to try and track how rapidly they release larvae, how many larvae they’re producing, and potentially allow us to videotape the process. It would take a bit of luck because we would have to put the oysters in individual beakers and hope that a few of them are brooding!
There are a lot of relatively inexpensive waterproof endoscope options on amazon. Ones similar to this look like they might work:
Took Laura’s K-treatment oysters to UW today, and discussion started about what projects I might be interested in doing this summer. We put the oysters in separate containers by temperature and OA treatment in warm salt water after being in a cooler all night to see if they would open up or spawn. We saw some activity, but are going to leave them overnight and then Steven will separate them all out into individual beakers tomorrow morning. Hopefully we get lucky and some were brooding and we see some larvae! We are hoping to do some experimenting with how I could best track spawning time and larval release.
I also spent some time researching small cameras that are used to record activity underwater and maybe even inside the oyster. Endoscopes are what have been used most often on bivalves. Specifically with investigating food movement throughout the gills and just general movement of particles taken in from the water. They come in a variety of sizes/quality.
After learning the full routine of screening, sampling, counting, restocking, and then collecting all the extra larvae to be frozen last week, things went much more quickly today. Laura was doing a lot of rearranging to make sure all her larval buckets are getting the same quality of food and are under the same conditions. I collected any new larvae that had spawned to screen through 100um and sample for counts. We got all the rearranging and restocking done before lunch!
After lunch Laura and I discussed possible research projects for me to work on this summer. Some of our ideas included:
-Histology + larval counts and how long does one oyster spawn
-Respiration experiment, metabolic difference
-Observing new larvae size differences from different treatments
-Mechanical stress or feeding experiment + mortality rate
We were very interested in discussing the project concerning the observation of the new larvae. Possibly using imageJ to track size differences and growth. This would be interesting because this work has been done on Pacific oysters, but not Olympic oysters who brood their young. It also would contribute to Laura’s project as well.
I was sent home with 15 oysters from each of the 4 treatments from the K-group that I will take to UW on 6/1 in case I want to use those for my project.