Take Two

1 02 2014

Recap: I am a Master of Science (and the Universe, according to my nephew when he wants the tickle attacks to stop), and my experiments in the intertidal were mostly successful, except for the live spawning attempt.  Again, to validate my project design and strengthen my simulated spawning results, we tried a second live spawning experiment at the end of last year before the permits expired… 

Me showing some abalone love (B. Blaud)

Me showing some abalone love (B. Blaud)

San Nicolas Island is one of my favorite places on earth, if you haven’t picked that up from my previous posts.  I feel incredibly blessed to have the unique opportunity to venture back, even after my schooling is complete.  I was plotting ways to get back to SNI, thinking of leveraging my experience on the island and in the intertidal to get a volunteer position with another group, when I got an email from Glenn that we were on for another live spawning experiment, and they would like me to be involved.  I didn’t even need to think about it before quickly replying, a hearty and enthusiastic “YES!  When do we leave?”

 

Withering syndrome is still a concern, even though it doesn’t appear to be as virulent as it was in the 1990s during its initial appearance on the island.  Although the disease presence has been documented on the island, the conservative approach to preventing further spread of withering syndrome dictates using only disease free animals.  The last spawning experiment involved naïve animals that were never exposed to the disease.  However, the difficulty we experienced spawning the larger animals and the learned knowledge that smaller animals are more gravid and easily spawned encouraged us to get younger red abalone.  To stay within the permit guidelines and only bring disease free animals to the island, the thirty red abalone we got for the experiment had to be treated with oxytetracycline (OTC), a broad spectrum antibiotic active against many types of bacteria, including rickettsiales.

 

The most ripe red abalone with bulging gonads were hand-picked by an abalone expert, Jim Moore, to improve our odds of a successful experiment.  Unfortunately, the OTC treatment is rather stressful, and involves the abalone sitting in a series of eight baths for 24 hours each over the course of three weeks.  During this time, the horny little abalone reabsorb their gametes, or may spawn in response to the stress.  To reduce their stress and hopefully maintain fuller gonads, the abalone were fed during the treatment.  They were allowed to recover for an additional three weeks after treatment concluded, and we hoped it would be enough.

 

Armed with a greater number of smaller abalone and knowledge from what led to failed experiments previously, we headed out to the island.  I think I inadvertently cursed myself after my last blog, Failure in the Field:

It is common for the first, and even the second field experiment to not work.  Unfortunately, I had everything riding on this one spawning release. There were many things I learned from this venture, the most obvious being to mix the iodine into the Lugol’s solution before adding it as a preservative to the eggs (duh).  I also later learned that the larger abalone are more difficult to spawn, and more successes are achieved with younger, smaller abalone.

In this second field experiment, I was in no way about to be in charge of the Lugol’s solution, and we brought formalin as a back-up preservative, just in case something else went wrong.  It’s science, and “things going wrong” is the name of the game. 

 

Red abalone in 3L of sexy spawning cocktail (Tris and hydrogen peroxide) (B. Blaud)

Red abalone in 3L of sexy spawning cocktail (Tris and hydrogen peroxide) (B. Blaud)

We set up the abalone in the temporary housing outside the archeological labs and storage, and less than a mile from a good release location at Site 2.  The plan was to spawn the abalone in the comfort of civilization, near electricity and running water, then take the gametes down to the experimental site, run the experiment at Site 2, and transport the canisters back to the lab where we would distribute eggs among the transportable tubes and add preservative.  That was the plan.

 

Now we just need the little buggers to spawn!  We added the sexy cocktail of Tris and hydrogen peroxide, ran down to quickly set everything up, and then waited… and waited… and waited…  When the tide started coming back in, and only one female had spawned a small amount of eggs in 6 hours, we decided to call it a day.  We put them all back in their temporary tank, and prepared to try again the next day.

 

Unfortunately, we were out of 6% hydrogen peroxide.  Luckily, someone thought ahead and brought back-up 3% hydrogen peroxide, purchased for $0.89 a bottle at the local pharmacy.  We doubled up the dose to try, try again.  Feeling slightly less optimistic, but still hopeful, we tried all the tricks we had learned in our combined spawning experiences and reading: temperature stress (exposure to sunlight to increase water temperature to 20°C in their 3L buckets, then water changes to bring the temperature back to 14°C); mechanical stress (EARTHQUAKE!  Shaking the buckets, tapping the buckets, mainly just agitating the poor abalone); musical stimulation (Barry Manilow, Madonna, Bruno Mars, Rhianna and Brittany – don’t judge!  It worked last time, and I was willing to try anything).  Our efforts were rewarded with some stressed out abalone that released several mucus plugs and poop, and appeared to want to spawn but had nothing to give.

 

Our conclusion: they hadn’t had enough recovery time after the OTC treatment.  Next steps include renewing the permits, applying for grants, and praying that the third try does the trick.  On the bright side, I get to go to the island at least one more time.  Can’t complain about that!





Invincible Black Abalone

29 12 2012
Pre-disease black abalone population on San Nicolas Island (G. VanBlaricom)

Pre-disease black abalone population on San Nicolas Island (G. VanBlaricom)

When I first began this blog, I introduced withering syndrome and it’s devastating effect on black abalone populations in Southern California.  The effect of the disease on the populations on San Nicolas Island is the equivalent of wiping out all the people in the world except for the populations in the US, Indonesia, and Brazil.  Only three of the most densely populated countries out of the total 242 countries in the world manage to survive.  It’s a little sobering when you think of it that way.  If you break it down so populations are evenly lost among each country, only the populations in Washington and Idaho would survive in the United States (I’m in Washington, so I’m happy I survive the fake apocalypse).  I’m not stating the statistics in the manner to convince everyone to move to the northwest, even though it’s the best place to live, I’m just trying to paint a picture of how the US would look if only 1% of the population survived.  Out of the 315,077,987 people in the US, only approximately 3 million make it.

 

Then, imagine among the people dodging the zombies in the apocalypse (I’ve been watching a lot of Walking Dead, so that’s the only apocalypse scenario I can picture right now), there is a group of individuals resistant to the brain-eating bacteria!  And they’re found on San Nicolas Island.

 

Disease-resistant black abalone discovered on US island

Biologists have discovered that black abalone on San Nicolas Island in the Santa Barbara Channel are more resistant to the deadly bacterial disease known as withering syndrome than abalone on the mainland. The discovery may help save these now rare intertidal molluscs from extinction, as scientists hope to soon breed these animals in captivity for release in the wild.

According to Carolyn Friedman, from the School of Aquatic and Fishery Sciences at the University of Washington in Seattle, San Nicolas Island has been the site of several severe outbreaks of withering syndrome. The present population appears to be the descendants of that 1% of the population that survived the onslaught.

Friedman and her colleague on the California Sea Grant project, Steven Roberts, also at the University of Washington, are now trying to identify which genes are responsible for resistance and the mechanisms by which this resistance is conferred. This work includes studying differences in gene expression between island black abalone and those from Carmel in Monterey, as the animals are subjected to high loads of the withering syndrome pathogen.

Withering syndrome, which causes severe atrophy of the animal’s foot muscle and is caused by a water-borne pathogen excreted in abalone faeces, occurs in relatively warm water, such as those found in the Santa Barbara Channel. Until recently, waters off the more northerly Carmel have been too cold to trigger outbreaks. As a result, abalone in Carmel have little natural protection against the disease.

Source: California Sea Grant

Anonymous.  2008.  Disease-resistant black abalone discovered on US island.  Marine Pollution Bulletin, 56(1): 7

 

Super Black Abalone (B. Blaud)

Super Black Abalone (B. Blaud)

Beginning in 2001, isolated populations on San Nicolas Island began to increase, so Dr. Friedman hypothesized that there may be disease resistant animals.  My very first trip to the island in 2004 was to collect animals and bring them back to UW to test this hypothesis.  The exposure of naïve (uncontaminated, un-diseased) black abalone from Carmel and comparing the rate of mortality to infected animals from San Nicolas Island indicated that San Nicolas Island black abalone are, in fact, resistant to the disease.  It’s a snail…it’s a rock…it’s SUPER BLACK ABALONE!  Yeah, I’m a dork.

 

But the story doesn’t end here.  Continuing this work, Dr. Friedman and PhD student Lisa Crosson investigated which genes were potentially responsible for the disease resistance in black abalone populations, and in doing so, made a surprising discovery.  But that is a story for another time…

 





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!





Spawning By Myself…

20 12 2012

Recap:  To bring more credibility to my project and determine how realistic my experimental design with surrogate particles is, I will release live gametes and measure the amount of fertilized eggs in a mesh container as a function of distance from live sperm, released from a fixed distance using an IV bag.  To learn how to spawn abalone, I participated in a pinto spawning up in Mukilteo several weeks ago.  I received 16 red abalone from the Cayucos Hatchery, and have been holding them for six weeks with the intention of further honing my spawning skills…

 

My red abalone (B. Blaud)

My red abalone (B. Blaud)

Spawning by myself sounds like something completely dirty, but I assure you, it’s not.  Since November 6th, I have been holding red abalone in the basement at the University of Washington with the intention of practicing the spawning protocol I learned with pinto abalone up in Mukilteo and honing the methods to something I can use in the field as part of my experimentBy the time I was ready to practice spawning them, I had 13 red abalone, 9 females and 4 males.  I was preparing to go to San Nicolas Island (SNI) for one of my experimental runs with surrogate particles, but wanted to have one solo spawning under my belt beforehand.

 

I headed down to the basement fairly early on the Friday morning to set up for the spawning experience.  The first steps are measuring up the participants, their gonads to be exact.  I measured the gamete index for each abalone, to identify which abalone was ripe, and separated them into buckets based on gender – boys in one bucket, girls in another.  Determining ripeness was a little more difficult than I anticipated, as none of my abalone were all that ready to spawn.  I looked under each of their skirts, but had a hard time identifying males from females in many cases.  It’s supposed to be fairly straightforward.  I hold the abalone, shell down in my right hand with the base of the swirl in the shell, their head near my fingertips and the apex in my palm.  After the abalone calms down for a second, I’m able to move the foot to the side to see the gonad at the base of the shell.  The gonad index identifies ripeness based on color and the amount of swelling in the gonad.  Yellowish-creamy white means male (makes sense – sperm is white), and a greenish-purple indicates female (the eggs come out an olive green).  An index of 0 is unripe, where the gonad is indistinguishable as either male or female; an index of 1 indicates when there is slight coloration and swelling of the gonad; an index of 2 has swelling of the gonad up to the mantle, the edge of the shell; and the highest index of 3 is visibly swelled over the edge of the shell, where you can see it with barely moving the foot to the side.  All of my abalone were at my un-expert opinion a gonad index of 0 or 1. 

 

Diagram of abalone gonad (Rogers-Bennett et al. 2004 J Shellf Res 23: 553).

Diagram of abalone gonad (Rogers-Bennett et al. 2004 J Shellf Res 23: 553).

I separated the abalone I rated as a gonad index of 1 into separate buckets, then put a couple abalone rated as 0 into their own buckets, since I couldn’t conclusively determine their gender, but wanted to spawn more than a handful of my abalone.  I added the appropriate amount of Tris(hydroxymethyl) aminomethane (or Tris, for short) and 6% hydrogen peroxide, covered them in a tarp to provide darkness (and a little privacy), and then left them alone with only short, periodic checks for spawners.  I mentioned before that these are all unripe abalone with a gonad index of 0 or 1, so it goes without saying that I wasn’t overly optimistic about the possibility of getting any of them to spawn.  So, imagine my surprise when I went down after only 90 minutes of them sitting in the dark to find one male and one female spawning in separate buckets!  I was so excited, I barely remembered that I was supposed to rinse them off and put them in a bucket of clean seawater (with no chemical additions), so they could finish spawning and have usable gametes.  As soon as that was done, I ran up a couple flights of stairs to find Glenn, who would share my astonishment and simple joy at these unripe, unready abalone suddenly releasing their gametes.

 

Hatched abalone larvae (http://www.lib.noaa.gov/)

Hatched abalone larvae (http://www.lib.noaa.gov/)

Eventually, after about 3 hours, all the red abalone had been rinsed and placed in buckets of clean seawater.  Due to the unripe nature of the abalone, they didn’t spawn a significant amount of gametes.  It was a great exercise going through the protocol, and learned a lot about what would work in the field, what needs to be modified, and ponder what it would be like to be an abalone parent, with millions of children, somewhere out there… but I’m weird.

 

After spawning my red abalone, a good practice run, I felt oddly optimistic about future experiments.  I should say, cautiously optimistic, because when you say the word “experiment,” I get an ominous chill and view of everything that could potentially go wrong.  I am starting to get more and more excited about the results that experiments with live gametes will yield.  But that, my faithful readers, is a story for another day.

 

And next, I will tell you all about Run 7, and my latest stay on the island…I can’t wait to go back!





Project: Funded!

17 12 2012

After 33 days of nail biting, annoying spamming all my dear friends and family, and cajoling strangers into coughing up their hard-earned dough, my crowfunding experience has come to a close.  Although I didn’t quite reach my goal of $2,500, making $1,990 is nothing to cry about.  I am in awe of everyone’s generosity!  I couldn’t have asked for a better experience.

 

Black abalone on San Nicolas Island (B. Blaud)

Black abalone on San Nicolas Island (B. Blaud)

What next, you ask?  First off, I’m going to stop slacking on my blogging responsibilities.  I apologize.  Sometimes, when I have a lot going on in my life, I have to drop one or two balls that I’m juggling or else I risk them all falling and hitting me on the head.  I have had enough bruises from past experiences to learn that lesson the hard way.  So while I haven’t been updating you as diligently as promised, I have been busy, getting new material for posts and always, always, always working on my project.

 

Second, I’m going to spend my money!  I love shopping!!!  But my RocketHub funds will be responsibly spent on small, white particles the size of abalone sperm.  I just got back from SNI, where I had an amazing trip and completed a successful extended run, and hope to repeat that experience with particles I purchase using what you all generously donated.

 

Third, I’m going to put my spawning practices to use by releasing live gametes in tide pools and find out how realistic my project design is.  And then… a lot of data analysis and writing.  That’s when all the free coffee I get at work at Starbucks will really come in handy, because sleep will become an expensive luxury.

 

So, while this quick update has been short and sweet, I wanted to get a HUGE THANK YOU out there to all my incredible funders.  Seriously, thank you.thankyou





My Thankful List :)

22 11 2012

On this wonderful holiday, full of stuffing my face with turkey and pumpkin pie (oh, how I love you, pumpkin pie), I thought I would take a second to reflect over my last year and tell you all about what I’m really thankful for.

The beginning of the hike down Heartbreak Hill (B. Blaud)

 

Last year, on Thanksgiving, around this time of 11:14am, I was preparing to go out in the field on San Nicolas Island.  We were headed to Site 8, which involves a 2-mile hike to get to our chosen tide pool.  The hike to the site isn’t so bad, all down hill and passed hauled out California sea lions and elephant seals, a bit brutal on the knees but nothing to stress about.  The run itself went smoothly, which was a blessing following the devastating failure the day before.  The sperm-sized particles were clogging the tube and not flowing consistently, so we tried removing the drip trap and connecting the tube directly to the bag the night before.  It worked, and I had my second complete, successful run.

 

After the run was completed, all the samples collected and everything packed back into my backpack, we took turns carrying the burden back up the hill.  The first 1 ½ mile back isn’t back, but the last ½ mile involves a 600-foot climb, which is, in a word, BRUTAL!  To save my pride, I insist on looking for whales in the ocean every 100-feet or so (ok, sometimes every 50-feet.  Don’t judge!).

 

E.O.D.B. (B. Blaud)

Once we reach the top of the mountain, we celebrate our success and survival summiting Heartbreak Hill (as the hill was christened decades ago) with an End Of the Day Beer (EODB), kept chilled in a cooler of ice.  Nothing tastes as sweet as a cold beer at the end of a long day of fieldwork and good hike.  When we get back to Nick Town, I cleaned off my equipment and stored my samples for safe travel back to Seattle while the rest of the team started on dinner, and what a dinner it was! Turkey, potatoes, stuffing, cranberries, green bean casserole, warm rolls, apple pie, pumpkin pie, and lots of wine.  Yum!!!

 

 

Mike Kenner’s improvised turkey baster (B. Blaud)

This trip and the following trips were great opportunities following the loss of funding.  March and April were spent measuring particles in my numerous water samples and analyzing the results.  During the summer though, I hit a bit of a slump.  I was no longer doing fieldwork, all my samples were processed and analyzed, and my future fieldtrips were months away.  I became unmotivated and uninspired.  I was still faced with the same lack of funding, so circumstances looked pretty bleak.  Everything always works out though, and I know this!  I just had to hold on, keep working and doing my best, but sometimes, you lose sight.  To promote my project and let you all know what I’ve been up to, I started this blog.  In writing about what got me into researching abalone and what an amazing species they are, I started to get excited again.  I fell in love with my research all over again.  I describe this in my interview with Anthony in his Notebook.

 

 

SciFund – Black Abalone Dating Service

At a conference at the end of September, when discussing the situation I was in with a fellow researcher, they suggested I try crowfunding on RocketHub.  It was less than a week later that I received an email from SAFS about fundraising through SciFund, which is promoted on RocketHub.  Everything is worth a try, so I went for it.  This bring us up to present, Thanksgiving Day of 2012, and my thankful list:

 

Mainly, I’m thankful for my friends and family, who have unconditionally supported me.  You have cheered my successes; been my shoulder to cry on; offered advice, ideas, and support; read my writings and drafts; and most importantly, are there.

I’m thankful for all my fuelers generous support.  On RocketHub, I have raised $1,150 and am at 46% of my goal with 22 days left.  I’m confident and optimistic that we will get there!

I am in awe of the wonderful opportunity I have to work with an incredibly species in a truly relevant field of conservation biology and ecology.  I’m doing what I love, and someday (hopefully sooner rather than later) will have a piece of paper that says all of you must call me Master.  That’s what a Master’s degree means, right?

I should also mention that I’m thankful for this blog, for reminding me what I do and why, and why I love it.  Sometimes, we just need to be reminded.  It’s actually been a busy month so far with my project and the fundraising; so much so that I have struggled keeping up with my promised 3 posts a week, but will work on it and do better!

 

So in closing, I wish you all a safe and happy Thanksgiving!  May you all eat too much and spend it in wonderful company!





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…