Run #7

24 12 2012

Recap: My preliminary runs were exciting, although it was difficult to get used to releasing a thousand dollars in the form of small particles and watching it drift away in the ocean.  In my experiment, as designed, a single male abalone was incapable of fertilizing an egg, even if a female was spawning right next to him.  I wasn’t convince that my design was perfectly awesome since it only showed the results of a single male spawning for 10-minutes when often a spawning event takes place for up to an hour.  After revising my project design, I was ready for a second take…

My tide pool at Site 8 (B. Blaud)

My tide pool at Site 8 (B. Blaud

 

It was the perfect day for a tide-pool experiment.  The tide was low, but not too low, the sky was clear, the temperature a comfortable 72 degrees, and the waves were not too high.  By all reasoning, I should have been in a great mood, but I couldn’t relax.  The experiment has tripled in size, and we are now releasing three times as many particles into the water, which means that over $2500 is now being released into the tide pool to float away.  I couldn’t deal with any thought of failure and wanted this run to be successful so bad, I could taste it.  It tastes salty, like a combination of ocean, sweat, and tears. 

 

 

I scrambled to get everything ready: all the sample bottles clean and present, all the tubes labeled and secured, triple checked that the particles were packed (there’s nothing like hiking for an hour to realize the guest of honor is back in the room), and hoped I wasn’t forgetting anything major – I knew I would forget something, I could feel it, but was hoping it was something that could be fixed in the field.  While setting up for the run down at the site, I realized I forgot the snap-shackles, which are used to firmly anchor the nose of the IV tube in the water.  I knew I forgot something!!!  Annoying, but not major, and something that can be improvised with some well-placed rocks.  Catastrophe averted, but I still couldn’t relax.  There were too many possibilities for things to go wrong along the way.

 

Black abalone on San Nicolas Island (B. Blaud)

Black abalone on San Nicolas Island (B. Blaud)

I went over the choreography for the experiment, because in my mind, the whole thing is like one big dance.  It needs to be timed, everyone’s movements synchronized, and when it goes well, it’s quite pretty to watch.  Dave, to clarify after I described the process, asks if he is releasing his sperm at 0, 1, 5, 10, 20, 30, 45, and 60 minutes after the initial release.  Incredulous, and rather impressed that anyone would be able to release their sperm in timely intervals like that, I suggested that I be the only one to release sperm, as mine is in the form of surrogate, sperm-sized particles and not real sperm.  Everyone agreed that that would be best, and would collect samples at those intervals instead.

 

Everyone took their place, and I had enough amazing volunteers that each person was responsible for only one distance.  I was the bucket lady, running around rinsing sample bottles between collections to avoid left-over particles, making sure the sperm supply didn’t run out, keeping track of the time and orchestrating the sample collections.  Someone later suggested I should teach an aerobics class, because my voice barked orders in a no-nonsense way.  I took it as a compliment.

 

It all went rather smoothly.  I glared at Glenn when he suggested as much halfway through the experiment.  “Don’t jinx it!”  I know science and superstition don’t exactly go hand in hand, but science is about exploring the unknown.  I’m not about to tempt fate and will do anything to ensure a successful run, including not mentioning how well it’s going until I’m sure nothing can go wrong!  It’s along the same line as not saying the number seven at the craps table when the button is on.  If someone had suggested that I can also improve my odds of success by clicking my shiny, red heels together while exclaiming, “this WILL work” three times, I would have done it without a second thought, totally comfortable looking like an idiot as long as all is right in my world.  Essentially, even though it was going amazingly well, I still couldn’t relax until the last sample was collected and safely packed away.

Mel, Eric, and Glenn enjoying an E.O.D.B. at the top of Heartbreak Hill (B. Blaud)

Mel, Eric, and Glenn enjoying an E.O.D.B. at the top of Heartbreak Hill (B. Blaud)

 

Despite my fears and insecurities, it was perfect.  There are always things that can be corrected and perfected, but nothing major went wrong.  Once all my samples were stowed, I could finally smile and laugh at the sperm-related jokes.  At the end of the day, for the first time ever, I was able to hike up Heart Break Hill without stopping to “look for whales,” while carrying a 50lb backpack.  Although my backpack was heavier, weighed down with water samples, my burden was lighter now that the run was successfully completed. 

 

Since we were only able to find funds to pay for one run, the rest of the week was spent in the kitchen counting particles.  Planning for this, I brought a dissecting scope (I’m no longer going to even try to deal with a coulter counter) and all the supplies needed for counting the egg and sperm-sized particles.  With few distractions that life on SNI offers (mainly, no cell reception and limited internet access), I was able to buckle down and power through the counting, completing over half my samples.  It’s a good start!





MacGyver-ed Experiments

1 11 2012

If MacGyver were a marine biologist, I try to imagine what complicated experiments he could create using a roll of duct-tape, a shoelace, and an empty Coca-Cola bottle he found on the beach.  I have a strange imagination.

I tried to channel some inner-MacGyver while working on setting up my experiment.  Remember, I’m trying to figure out how close together two abalones must be in order to successfully reproduce.  I had a rough idea of how the experiment should go in my head, but sometimes it’s hard to connect what I envision with reality.

Tide pool used in experiments on San Nicolas Island (B. Blaud)

So here’s what I initially pictured: I would let a sperm cloud build up for five minutes using a controlled release from an IV bag, commandeered from a local hospital by sources unnamed (love you big sis!!!), then I would release the eggs all at once and collect water samples.  This seemed pretty simple and straightforward in my head until my advisor, Glenn VanBlaricom, asked how I would collect the samples.  We brainstormed different systems, one involving placing 10 people in the tide pool with bottles, scooping up water samples, and another with sampling tubes rigged onto a frame that would be anchored to the bottom of the tide pool.

Sampling bottles (B. Blaud)

What we ended up doing was something in between the two ideas.  I thought it would be useful if the sampling bottles functioned similar to a Niskin bottle, which is used in oceanography to take water samples from different depths.  It is hollow and open on both ends, which allows water to flow through.  Water samples are collected at a desired depth by triggering a mechanism that allows the stoppers on either end of the tube to close.  These bottles range from a couple hundred to several thousand dollars, are much larger than I need for my samples, and are entirely out of my budget of $52.37 (all that I had in my checking account at the time).

With the invaluable help from Julia Eggers, I was able to create unique sampling bottles using PVC pipes, racquetballs, surgical tubing, and zip-ties.  Eggers and I spent a couple afternoons scouring the local hardware stores and harassing poor Home Depot employees to figure out how we could make this contraption hold water.  After collecting supplies, we would run back to the shop at school and experiment with different designs.  Through trial and error, we came up with something pretty darn cool and simple.  To make them in bulk (I needed 40 of them), I got all the supplies together, bought pizza and beer, and invited friends over for an assembly party.  Ah, what my friends won’t do for a free beer…

Chris Yates with a water sample that has visible egg- and sperm-size particles (B. Blaud

To get the whole thing to work, the simplest plan was to place actual people into the tide pool at the time of the experiment, and run it like I had envisioned.  We had a couple other experiences in the trial and error process that were fairly painful.  To get an idea of how the experiment would go, VanBlaricom and I headed out to practice in a tide pool.  We set up an IV bag with the fake-abalone sperm and put the fake-abalone eggs into the water using a turkey baster.  One problem we didn’t anticipate: the particles floated – and so while we watched, literally $1,000 floated away into the sunset!  I could have cried, and maybe did a little.  All my research online indicated they would be neutrally buoyant in the water, but we totally discounted needing a dispersant, something that would break the surface tension of water and allow the particles to enter the water column.  Dawn detergent to the rescue!  The next day, we headed back to the same tide pool with Dawn and another batch of particles, and this time we were successful.

IV bags, the right one containing sperm-size particles (B. Blaud)

For the short experiment practicing to see how the particles would react and brainstorming how we would choreograph the entire experimental run, the IV bag worked great!  Unfortunately, in the extended, real runs, it clogged (another $1,000 floats off into the sunset with no data collected – more tears shed, and not the last).  I used swiss army knife to remove the drip chamber where the clog was occurring and attached the tube directly into the bag with duct tape (see, I AM like MacGyver, solving the world’s problems with a piece of duct-tape!).

I set up a tripod to hold the IV bag, filled it with fake-abalone sperm mixed with seawater and Dawn detergent, and let it release for 5-minutes before releasing the eggs.  Everyone had his or her own location, a set distance from the egg release location and collected water samples at precisely 0, 15, 30, 60, and 300 seconds from the time the eggs were released.  The choreography was something else that was worked out through trial and error, but after several runs (some successes, some failures), we got a good system worked out.

And thus, we completed six successful runs of simulated spawning experiments, providing me with roughly 684 samples to run process.

Next up, from the field to the lab…