Post #14 - What is it?

It's been a busy day in the lab, but the afternoon ended with a quick snorkeling trip.  Saw some things I thought you'd like to see, though they are presented here through a serious of questions.....

(It was a late afternoon snorkel, with threatening clouds.  If fact, it was raining by the time we were getting out, and pouring later.  Because of the time of day and weather, the colors in these pictures are rather muted.)


First question: This is a close-up of what?

The top side of a cushion star, which like the COT eats corals.  This one is about 6 inches across:

Second question: What kind of animal produces all this mucous coming out of a hole (at least 4 of which you see here) in the coral?

It's hard to believe it's a snail, but not at all like this cowry which crawls about freely:

The mucous is a sticky trap for organic rich particles that are reeled in and eaten (along with the mucous) by vermetid snails.  The shells of these animals look more like the calcareous tube of some fan worms, and are usually buried down in the coral skeleton but not here:

The round dark "trap door" is the operculum, just like that of most free-crawling snails that block the opening to the shell should a predator arrive.  In this full-sized animal, the operculum is just less than an inch across.

Third question: What did Emily take a picture of here?  An eel?  A snake?  A worm?

Nope - it's a sea cucumber, at least the back 12 inches of it.  If you touch it it feels "sticky" because tiny barbed ossicles (little bones) protrude from the skin.

Lastly, see if you can find Waldo, the 3 inch baby flounder:

Post #13 - 3 x 3 - we did it!!

Today ended with a bit of a celebration.  Our goal for the COT fertilization study was to cross each of three female Acanthaster with each of three males, and today we ran the 9th experiment!    We still have tubes of embryos and sperm to analyze, but I should be able to do that in the next couple of days.  Then, being forever curious, we get to decide what additional crosses we should do while we have the opportunity.

I have told you a little bit about the fertilization experiment in bits and pieces,  let me try to present a more complete picture.  After removing some testes and ovaries from adult Acanthaster, I collect the eggs and sperm and make separate suspensions of known concentrations.  (I have to be really careful not to get any sperm in with the eggs for that would ruin everything!)  This is what the suspensions look like:

The dots in the right tube are eggs, each being about 160 um (that's 0.160 mm or 0.0063 inches).  I'll show pictures of sperm some other time.  The actual mixing of eggs and sperm occurs in the aquarium building:

On the left is a locker for dive gear, and inside the gray/white box on the right is a chiller that helps regulate the temperature of the water in the aquaria.  Around here, we call the aquarium room the "Aqualogic" room because of the company that makes the system.

For each aquarium, the system allows us to control water temperature by using both heaters and chillers,  and the pH of the water by bubbling carefully controlled gas mixtures.   In an adjacent room is the equipment for controlling the mixture of air and CO2 that gets pumped into each aquarium:

The six aquaria on the left have just air bubbling into them, while the tanks on the right receive air enriched with CO2.  As you know, since the CO2 dissolves in the water forming carbonic acid, the right tanks have a lower pH (are more acidic) than the left tanks.  In addition, for Emily's experiments half of each set of tanks also have a slightly higher temperature.

Water from the tanks is filtered to remove bacteria, then pipetted into a series of culture tubes.  This can be done with or without feline help.

Each run involves eight tubes, plus three more for pH measurements.  A serial dilution of sperm is made in the eight tubes, designed so that the first tube has way too many sperm (causing some eggs to be fertilized by more than one sperm and develop abnormally) while the last tube may not receive any sperm at all (resulting in no fertilization).  The reason for doing this is that ocean acidification may affect fertilization success in different ways, and these ways might be revealed by using such a huge range of sperm densities.  Once the tubes have their sperm, then approximately the same number of eggs are added to each:

The tubes are floated in one of the aquaria, as seen in a previous post, for six hours.  At this point, the embryos are preserved.  When examined under the microscope, the results are pretty cool!

All the embryos in the row at the bottom are normal blastulae (hollow balls of small cells), except for the second one from the left that is abnormal, presumably from polyspermy (too many sperm entering the egg).  The embryo at the top is also abnormal.   For every experiment, one set of tubes is filled with regular sea water, while another set of 8 is filled with high pCO2 water.  The results from both series will be compared to see if the more acidic sea water affected fertilization.

So even though there is work yet to be done, having actually finished running our 9th cross was a good reason to celebrate - at a nearby creperie!

Post #12 - What a productive day gets you...

It's a great way to start day when first thing in the morning you check the pH in the aquaria and the levels are what you need for experiments!   Today was such a day, so we did two fertilization experiments.  The first one started about 8 this morning (after an hour of checking the pH levels) and the embryos from the last run were preserved around 7:30 p.m.  So this is what my lab bench looked like at the end of the day:

The test tubes in back and in the blue box contain the embryos and/or unfertilized eggs that have been preserved in formalin and need to be analyzed.  The smaller vials in the white foam block (scrounged from a packing container) hold preserved sperm suspensions, also waiting to be counted.   As of yesterday I was all caught up, but all these tubes will take some hours to go through.  However, since they are preserved I can work at them whenever there is a free moment.  For tomorrow morning, however, another experiment is planned.....

Late this afternoon, while the culture tubes were incubating, Emily and I went snorkeling to scout out coral colonies that she will be collecting tomorrow.  I took my camera along and thought I'd share some of today's pictures:

Here's a closeup of Pocillopora damicornis, the species of coral Emily is working on.  The width of this picture is about 4 inches.

Next to this coral colony was a colorful colonial anemone (also showing about 4 inches ):

Tropical fish can be spectacularly colorful, such as this box fish seen here:

Curious people constantly ask questions, trying to understand what they observe, and graduate students are no exception.  I spotted this not too far from the SeaFET.  It's a rack with pieces of coral wire-tied to it.  (Plastic wire ties have a gazillion uses!)  A CSUN graduate student that left last week is trying to determine how "plastic" the growth form is of this species of coral.  It can form finger-like projections on the upper parts of the colony, but has a more plate-like growth form around the lower edges.  She is transplanting pieces of colony to different locations on the colony to see if they grow in a manner appropriate for their new location, or retain their original growth form.

It will be interesting to see how these bits of coral grow....

Post #11 - Bumpy road

The island's roads have their share of bumps, but on average are pretty smooth.  The same can be said for our avenue of research here, though for the past few days the "road" has seemed like it's full of potholes!   The aquarium room has 12 tanks, and for each we try to control the temperature and the amount of CO2 bubbling into the water to influence the pH - the more CO2, the more acidic the water becomes.  (I'll post pictures of the tanks later....)  We can't run our COT experiments if the pH is not within certain limits, and Emily also needs to have some of the tanks at a slightly higher temperature for her coral larvae experiments.  (Her corals should begin releasing their larvae after next Wednesday's new moon and continue for about 7-10 days, so we need to have everything working before then.)  Well, two days ago the high CO2 tanks' pH values were off, and the chiller that helps regulate the temperatures wasn't working right - so trouble shooting all day and no COT experiments.  Yesterday, the pH values were good so we ran TWO COT experiments, starting in the lab around 7 a.m. and finishing preserving the last tubes of embryos at 11:30 p.m.

We had planned on running another COT experiment this morning, but alas, the pH values were off again because the air mixture pumps shut off unexpectedly overnight.  After the pumps were working again, it took until this evening for the pH values to get to where we needed them.  So today was another day of no COT experiments, but of spending hours looking through a microscope analyzing the embryos of the past few experiments, making a very quick SCUBA dive to collect a water sample next to the SeaFET, installing another chiller in the aquarium room, doing many pH measurements, and getting ready to do two COT fertilization experiments tomorrow assuming the pH values in the tanks will be where we need them.

There's progress every day, it's just not always what is planned!

Enough of that....  Here are some pictures for you:

These COT always amaze me!

Collecting carefully!  (My finger that was injured over two weeks ago is mostly healed but still sore.)

Coral reefs are cornucopias of colorful animals.  Here is a "giant clam" (this little one is about 4" across) with its mantle (the tissue that secretes the shell) a beautiful blue.

However, clams, like people, show variation......

One of my favorite animals is the nudibranch, sometimes called a sea slug.  These can be spectacularly colored, and are fun to find though at times they can be very cryptic.  However, this 6" guy was not hard to spot; it was right out in the open.

All the pictures in today's post were taken a week ago by Viv Cumbo, a post-doc from CSUN that graciously shared her great pictures with me.

Post #10 - Toothbrushes come in handy

Today was an interesting, diverse day.  The major effort was to get our first "official" COT fertilization experiment underway.  We are confident that we have worked out the major bugs in the protocol so it's time to get some data!!  The morning was busy checking the pH of the water in the aquaria we were going to use, collecting eggs and sperm  and making the suspensions of the right concentration of each, etc.

We got the gametes into the culture tubes by lunch, and since the tubes are incubated for 6-7 hours suspended in an aquarium, we felt after lunch would be a good time to check on the SeaFET.  Tomorrow we need to collect water samples right next to it when the device is taking its measurements of pH, etc.  (Emily's watch is synchronized with the timer inside the SeaFET so we know when it sampling.)  As we headed down to the dock to prep for the snorkel, I asked Emily if she remembered her toothbrush.  "Of course, dad!" was her reply:

Toothbrushes are wonderful devices for cleaning things, from "les dents" to screens around SeaFET sensors:

We all know how easy it is to ignore things around us when you are concentrating, as Emily did here as an inquisitive moral eel looked on as she brushed...

As I said in the previous post,  you never know what you might see when out in the water.  There is always the possibility to be surprised.  Today, again, was no exception:

The eagle ray was just cruising by...

then it was off to wherever it was heading...

as Emily said goodbye....

(She's not touching the ray, but is probably within 5 feet of it.  A magical moment...)

Post #9 - Rays and stars

Today started out gray and wet (it is the rainy season here, after all),  and the morning was spent conducting another trial run of the fertilization experiment.  It's critical that I get the right sperm concentration in my culture tubes, and that is turning out to be tricky.  The sperm concentration of my "Sperm Stock Solution" is determined by using a special slide called a hemacytometer on which I count the sperm found in a tiny volume.   Inexplicably, for the last two days, the sperm counts in my stock solution have been significantly HIGHER at the end of the experiment than at the beginning!  "Go figure!"  I assume it has something to do with my technique so I am contemplating what I can do differently.

An after-lunch excursion was planned for collecting more COT among the "bommies" (small coral heads) at the northwest corner of the island.  Such trips always fill me with anticipation, for one never knows what you might see when out in the field, and this afternoon's trip had several surprises.  Vinnie was our boat driver.  He's a technician from California State University at Northridge ("CSUN") here with a group of grad students and a post-doc.  They all leave Tuesday and since he knows the tricky path through all the reefs to get to "COT city,"  I wanted him to show me how to navigate there one more time.  However, along way he wanted to show us a couple of different spots, completely unrelated to COT.  The first simply looked like an area of patch reefs, but in reality what looked like coral heads were actually sunken stone Tiki carvings.  I am not sure how they ended up in the lagoon but one rumor is that the artist that carved them was so dissatisfied with the final products that he dumped them in the water.  (Not an easy thing to do considering their size...)  Regardless, it's a popular snorkeling spot for tourists and COT researchers alike:

Here are Emily and Viv, the post-doc from CSUN, not quite succeeding in mimicking the stone faces:

Although it was interesting to see underwater stone carvings, I was much more enthralled by our next stop, a spot where people frequently feed the sting rays so that they come flying in when they hear the anchor hitting the water.  It was enchanting to have about eight of these beautiful animals cruising about so elegantly, so gracefully. . . looking for a handout no doubt!

Then on to our COT collecting site.  I spotted the first one within 20 yards of where we anchored...

... and soon had reached my bag limit! (I could fit no more than nine inside.)

As a final picture, here is an inside-the-bag look of the COT ready to help push back the frontiers of science....

Post #8 - Plastic plates and a Leatherman

Question of the day: how can plastic salad plates and a Leatherman (which are like Swiss Army Knives) help advance science?   Here is the first picture to help you get started:

You are probably wondering what type of hints I'm giving you with the nine circles drawn in red ink and the one hole cut into the plate.  Any guesses?  Remember, I'm working at a marine lab.

Let me give you another hint:

Now that you see the culture tube next to the plate, it's obvious what this is for, isn't it?  Well, maybe not.

Recall that I'm studying the effects of ocean acidification on fertilization success in the COT. Ocean acidification is caused by the increased amounts of CO2 spewed into the atmosphere from humans burning fossil fuel, some of which dissolves in the oceans forming carbonic acid, which lowers the pH.  This process is very temperature sensitive, so when we run experiments, it's important to conduct replicates at the same temperature.  Therefore we can't do this out in the lab air, for that can vary by several degrees.  Thus we decided to conduct our fertilization experiments in tubes suspended in aquaria where the water temperature is carefully controlled. The challenge was to construct tube holders that we could float in the aquaria.....voila!

The Leatherman did a great job while the salad plates were available on this remote island and fit our budget!  Being a productive scientist often requires one to be resourceful!  Now our gametes will be kept at just the right temperature suspended in the aquarium water, as seen in this picture is taken through the front of the aquarium.

Post #7 - Fingers and fruit

Friends and family have asked how the fingers are doing, so here is an update:

I stuck my right finger onto a COT spine during my first encounter three weeks ago.  I've lost two layers of skin this past week around the healing wound, which is good.  There had been a large lump of what felt like scar tissue inside, so I'm glad it's gone.

The left finger was injured a week ago, and though it may look nasty, actually it is almost pain free.  I assume I'll be sloughing off skin from this finger soon too.

Super busy day in the lab trying to run some experiments, but I took time to take these pictures of the fruit tree outside our lab door.  Locally they call this "pamplemousse" which is the French word for grapefruit.  However, I've been told this is not a true grapefruit, but a relative in the pomelo family.

They are ripe when they easily twist off the tree, and delicious!  A bit sweeter and less tart than the grapefruit I get back home, and a bit more "connective tissue" around the flesh.  But they are fresh and available anytime!

Post #6

I took my sore finger (see Post #5) to the doctor this morning and with Emily's limited but much-better-than-my French speaking abilities, with a script of questions in French compiled yesterday with the help of a post-doc from France, and with the doctor's limited English abilities, things went rather smoothly.  Besides getting prescriptions for amoxicillin (to thwart bacterial infections) and cortisone (to reduce the swelling) I learned that the Tahitian name for the COT is "taramea."  (Pretty-sounding name for such an animal!)  Wonderfully, the doctor's visit cost only 3600 French Polynesian Francs (about $40US) and medicines only about $30.  

With the new meds in my bloodstream after lunch, I tried unsuccessfully to catch up on some much needed sleep, so I headed down to lab to solve how to reliably get COT sperm suspensions with a predictable concentration.  But instead of boring you with those travails, I thought I would share with you some pictures of the day.  (But just in case you are still wondering, the COT-stabbed finger is beginning to look and feel better tonight.  I'm glad, because it's real slow typing without it!)

Our seeming tranquility is altered by visitors from elsewhere that arrive using different modes of transportation.  This vessel anchored out front of the lab for less than 10 hours today, and missed the reef when it left!

Some would rather tour the island in an outrigger canoe:

Which experience would you prefer?

Every marine lab I've been to has a volleyball court.  Here's the one at Gump:

You can see from the intact grass that it's not used much, only once in the past two weeks.  Perhaps it's because folks here are too busy, or because it's too hot, or because the scenery is too distracting, or (most likely) because the only ball available is a tattered soccer ball.

So why not go scuba diving instead?  Here's the group of people that work in the same lab rooms as I do.  They are mostly grad students with one technician and one post-doc.  (A great group.  More on them some other time.)

They went across Cook's Inlet to a "wall" of a coral called Porites rus.  While there, Emily took these pictures of a beautiful nudibranch (a sea slug or shell-less snail)...

and an octopus peeking out of a hole...

with one eye showing above its white siphon.  I see octopus on most of my dives here; glad Emily spotted this one.

And when Emily came back from her dive, she just had to capture the full moon rising.  The resolution isn't great here because it was taken using her underwater camera in a housing not designed for above-water photography.  Still, you can "get the picture":

Around 8 p.m., the moon was our "street lamp" as we made our way through the forest and up the steep hill to our bungalow.