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.
I spent my first day at Manchester helping Laura screen and count her new larvae. After an initial walk through, Laura taught me how to screen, sample and count the contents of each bucket.
We started off sampling larvae that had been screened through 160um, 140um, 120um, and 100um with the goal of collecting growth rate data. Laura did the screening and I did the sampling and counting.
-I was given tripours containing the larvae from each size screening (4 tripours/bucket) that I filled to 200mL
-I took triplicate 0.5mL samples (dense amounts of larvae) or triplicate 1.0mL samples (less dense).
-Fixed wells with Lugols and recorded counts.
After a couple buckets we changed tactics and Laura decided to just screen to see if larvae were above or below a designated size. Since they all are pretty small right now we just screened the buckets through 100um screens to get total counts. Almost no larvae in the initial buckets were over 160um.
-Screened new larvae through 224um screen onto sorting table filled with FSW.
-Placed 2x100um screens under sorting table- top one clean and vortexed.
-Drained sorting table onto 100um screen.
-Collected larvae in tripour.
-Labeled with tape, left on bench while I collected the other groups.
-Vortexed sorting table & screens between groups.
-Sampled triplicates from each group, but did not fix with Lugols because we wanted to make sure we were able to distinguish between alive and dead larvae!
My computer really didn’t like dealing with this much data, but I started organizing it all into pivot tables and it seemed to work?
1. First I created a pivot table that summed all the rows labeled with the same protein name giving me one total number for each protein per site. I copied this data into a new sheet.
2. Then I summed each row and got protein totals across all the sites, which I then sorted by value from largest to smallest.
I put the file with my results in my folder on OWL:
Still working on figuring out how to upload the other documents.
Successfully made it to my first day at the Roberts Lab yesterday! I attended the lab meeting and logged into all the sites and platforms that I will need access to this summer. I left for the day with some readings to catch myself up on basic information pertaining to Olympic and Pacific Oysters. I also did a lot of browsing of Yaamini’s and Laura’s notebooks to learn about their current projects!