Spawn! But Just a Bit...

(Again, tried to post this last night, but didn't work.)















Well, we got a tiny bit of spawn tonight. Nothing like we expected though. One colony for us and a bit mixed from others and then 3 for Iliana's colonies. By trading some sperm with Iliana we were able to do our cryo experiments for tonight. Only two very light kreisels for our SECORE aquarists though. Hoping tomorrow is the big night.

Lots of pics today but I should go to bed so will post tomorrow.

Aloha!

Look at all the Volunteers! Wow!

Sandra's camera got a better pic of the whole group last night, volunteers and our group. Remember, there's only 9 folks in our group, so the rest are our wonderful volunteers!

Pics pics and more pics!

Since we got most of our stuff ready last night, today was a relatively relaxed day getting a few more things ready for the predicted spawning tonight.

Brian altered some of their collection nets.



Jason and Michael did some runs with the little freezer that Michael brought for us to try. Jason is entering some of the run data into his computer so they can see the rates that they were getting.



Michael and I were checking out the rates.



Mike shows us his altered collection net. All set for Mike's patented vortex method of collecting!



Michael with his freezer in action during a test run.



Mike and I geek out with our cameras. :)



Rafael does some test runs in his freezing box.



Possibly the most exciting part of the morning, Brian is using his aquarist skills and becoming my hero. I forgot to bring 5ml pipette tips for the trip, even though I brought and could really use a 5ml pipettor. Mary was able to get some long pipettes and I had Brian epoxy one to a syringe and now I can measure the amount of water I need for my fertilization vials. Woot!






A couple beach shots...




Looking back at the yellow house from the beach.



New critters for the day, hermit crabs at the edge of the beach. Very small and very cute!





Some underwater photography that Mike did this afternoon.






And the star of our show...



Michael and Jason ready for some snorkel-action this afternoon.



Had enough of a break in the afternoon I started putting together tracks for a new mix (surprise!). Notice the sunglasses with the headphones, rockstar style! ;)



Mary thinks the kitty might be brining us dead beetles as gifts as they are accumulating by the door.



Shortly before spawning time, our army of volunteers arrived. Sandra brought a ton of people tonight, which was great. They had everything covered for us and Iliana's group with two kayaks in the water to run potential samples from the divers to the shore, runners on the shore and back up folks all around. It was great having them here.



The volunteers listen as Mary explains what we need their help with.



Unfortunately, we got no spawning tonight. We'll cross our fingers for tomorrow. Everyone was dissapointed, but knew that this is how biology works, so we got out the drinks and some snacks and started to relax.






Then Brian got a call from Bob Snowden up at Iliana's house. Turns out one of their marked colonies spawned just a bit. Since it was just one colony, she couldn't use it as she needed to do fertilizations and they can't (or shouldn't...there's some evidence that it may happen, but offspring may not be viable) self-fertilize.



Mary ran up to their house and got the vial of eggs and sperm so that we could use the sperm to test some of our cryopreservation techniques. Here's Mary with her prize vial!



Mary gets the scope going so that we can see how dense the sperm is and how it looks, i.e. is it motile and intact?



Sperm looks really good at 2x10^8 (that's 200,000,000 sperm!) / ml.



Michael gets the freezer ready while Jason loads sperm into straws for some trials.




Rafael works on his faster freezing rate techniques.



Megan and Mary look at some thawed sperm and discuss its motility.



While it was dissapointing that the large group of volunteers didn't get to see a big spawn (some of them are able to return tomorrow) and unfortunate that we didn't get our full samples tonight, getting some sperm allowed us a nice opportunity to take some time to test some different cryopreservations. We were able to pin down specific cryoprotectants and rates that we want to try tomorrow night for actual fertilizations rather than having to do a ton of fertilizations that might not work. It was great that everyone got comfortable in the lab so we will be even more ready to go tomorrow night.

Good night for now!
Aloha!

We Wait...

(Originally tried to post this around 9:30pm while sitting on the beach, but phone wouldn't send it.)

The divers are in! The rest of us are waiting on the beach for the first samples to come in.

We have an army of volunteers tonight and Iliana's group is in the water too. It's a crowded night. You can feel the excitement.

Fingers crossed, we can only watch as the glowsticks dance in the water, hoping the movement is divers collecting not having to hunt for actively spawning colonies.

Think happy thoughts for the corals...
Pics from today to follow later tonight.
Aloha!

No Spawning Yet, but a Productive Day

Well, there was no spawning yet tonight, but we definitely had a productive day.

First we had a conference call up at Iliana's group's house with Mike Brittsan who is in Curacao with another crew of SECORE folks for the Elkhorn coral spawning there.

We had a nice lunch with our group at the Shipwreck Cafe. Nicole, who owns the restaurant catered for us two years ago, and is doing so again this year, but with such a smaller group we are able to go up there for lunch each day. She's a really great cook. Mary and I made a trip to the bank again today and another trip to the store to stock up on some food stuff.

The cryo-team arrived late last night. Dr. Jason Acker, from the University of Alberta joins us for the second year in a row. Being Canadian he's completely unadapted to the heat here, and as he says about last year "I don't think you've ever seen a human being melt before." Rafael Uribe is also joining us for a second year and comes from Dr. Terry Tiersch's lab at LSU. Megan McCarthy is new to the Puerto Rico team this year and she joins us from Stuart Meyers' lab at UC Davis. It's a really great group of people added to our already great group we have with the SECORE aquarists here. We had a really good meeting with our cryo-team to discuss our experimental plan and decide who was doing what. The meeting went well and I feel like we are well prepared for what we need to do. Briefly, Mary and I will be doing some optimal fertilization concentration experiments with fresh sperm to figure out what sperm concentration is best for fertilizing with this species. We will also be looking at a few different cryoprotectants to see if there is toxicity even before any freezing takes place. We will also be testing if the elkhorn coral sperm is chill sensitive. After all that, Rafael and Jason will be doing two different cryopreservation methods and we will be fertilizing with sperm that looks the best from that. Megan has the very important job of examining the sperm on the scope for density before experiments and motility before sperm comes to me for the fertilizations so we don't have unnecessary fertilization trials. This is the plan for the first spawning night and then based on our findings the next day (we can't analyze the fertilizations until the next day) we will form a plan for banking sperm the next night. I feel like we have a good solid plan this year and everyone has their roles and things should go smoothly. We got geared up this evening just in case we had an early spawn tonight, but they did not spawn. The actual spawn is predicted for tomorrow night, so no spawning tonight is not a bad thing. It was great that we geared up for it though because now we are all set for tomorrow night and just have to wait.

Eric, Brian and Mike have a really nice looking system set up in the bottom of the yellow house with kreisels for raising their larvae. They are having some issues with dissolved oxygen levels and evaporation, but some water changes tomorrow should alleviate the problem.

We also had some volunteers come this evening to see what we need help with. Steve Tamar volunteered with us last year and has been in touch with Mary all year and coordinated some groups to volunteer this year. He brought Joel and Sandra today. Joel is here interning with Surfrider Puerto Rico and is an environmental law student at Vermont Law School. Sandra is a professor at the University of Puerto Rico in Mayaguez and she coordinates an environmental program there. She will be bringing 9 volunteers tomorrow and 7 Sunday. Volunteers were a great help last year when I was on the beach trying to coordinate the divers and the collection of the spawn. The volunteers ran samples to the cryo team and helped get the samples from the divers. They also kept an eye on the houses and equipment when we were busy in the water and on the beach. This year we are on a much smaller scale, but the volunteers will be equally helpful. They will be split between helping with our group to bring us samples of egg-sperm bundles for our cryo work and our SECORE aquarists' kreisels and also helping Iliana's group and her SECORE folks.

Some pics from today:

We got the hose on the liquid nitrogen tank this morning and filled our dry shippers and our small liquid nitrogen dewar.



Getting the tank ready was pretty funny, apparently. :)



Mary is about to disappear in the vapor!



Dry shippers. These are neat little containers that have a material inside that absorbs the liquid nitrogen so that you can ship things that cold without having actual liquid sloshing around. Very useful!



The very nice set up for our group's kreisels this year.




Eric measures dissolved oxygen this morning and discovers it is quite low.





Brian snorkeling and fixing one of the bouy tags on one of our elkhorn coral colonies. Eric and Brian and Mike went out and picked coral colonies that looked good for our cryo-experiments. In order to do our fertilizations properly, we must ensure that we get eggs and sperm from different colonies, so marking good colonies allows them to go back with a next for spawning that will fit over a colony so that we know we get egg-sperm bundles from that colony only.



Discussing things with Sandra. Next to her is Michael Byers who has come to help Jason set up and use a very nice little freezer that can go at -15 degrees C per minute without the use of liquid nitrogen. Very nice of him to come all the way out here to ensure that we are able to use this piece of equipment to the best of its abilities. Mike Henley is playing on his iphone, of course. See pic below as well. ;) We are definitely the Mac crew here this year as Mary, Eric, Mike, Jason and myself all have Macs and then Mike, Jason, Michael and I all have iphones.





Mary, Rafael and Jason at our preparation meeting.



Megan also at the meeting.



Some cute little houses on a hill outside of downtown Rincon. Mary and I both love the use of color here. Wish we had more of that in Hawaii.



My fertilization trial vials all set up but nothing to put in them yet.



And our new critter for tonight. We have these in Hawaii too, but still not one I really wanted to see.

Aloha all!

P.S. Subphonix is tonight, so if you are reading this from Oahu, you should go. ;)