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!

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Allee Effect

22 01 2013
A "crackle" of black abalone on San Nicolas Island (B. Blaud).  Crackle: a group of black abalone.

A “crackle” of black abalone on San Nicolas Island (B. Blaud). Crackle: a group of black abalone.

The Allee effect was first described by its namesake, Warder Clyde Allee in the 1930s in an article on goldfish.  His article (which I cannot access but have a summary through other sources) investigates the effect aggregation has on overall fitness on the individual goldfish, in this case.  He found that goldfish grow more rapidly in a tank that holds more individuals.  Although he never comes right out and says, “the Allee effect is defined as…”, the generally accepted definition is that higher densities within a population lead to improved individual fitness.  This basically means that larger group sizes provide individuals with more opportunities to mate, defend themselves, feed themselves, and/or can work together to alter their environment in a beneficial manner to the whole group.

 

The term, Allee effect, was often misused and erroneously applied, however, so Stephens et al. (1999) decided to take it upon themselves and define the principle in clear terms laden with scientific jargon and information.  I have a secret to admit: when I see scientific papers, my eyes gloss over.  If the paper has equations, game over.  I check out completely.  If you talk to researchers, many admit to only reading the abstract (a roughly 200 word summary of the paper at the very beginning of the paper), looking at the figures, and skimming the conclusion.  With the overwhelming amount of publications out there, that really is the only way to see if the paper is relevant to what we’re studying and saves a considerable amount of time.  Between the jargons, big words scientists use to justify the tens to hundreds of thousands of dollars spent on earning advanced degrees; you can tease out the basic information of what they’re trying to get across.

 

Black abalone on San Nicolas Island (B. Blaud)

Black abalone on San Nicolas Island (B. Blaud)

Stephens et al. (1999) paper, titled “What is the Allee effect” has large amounts of jargon as well as confusing equations.  I was able to bear down and wade through it.  I need to justify the exorbitant amount of money being spent on my education as well, and prove that I understand demographic stochasticity as much as the next overly educated person.  Side note: I think that if my spelling autocorrect in Word doesn’t know that it’s a word, I should either.  But then again, the autocorrect doesn’t know my last name, so this opinion doesn’t have strong support.  But as usual, I digress; back to the point.  After exploring the numerous examples Allee lays out to support his principle, Stephens et al. (1999) is able lay out a clear definition:

 

…we thus define the Allee effect as: a positive relationship between any component of individual fitness and either numbers or density of conspecifics.

 

Basically, the more of you there are, the better you all will be in terms of certain basic needs (reproduction, food, protection, etc).  There are limitations with this.  Too many individuals and you become competitors for resources, such as food, so oftentimes there is a threshold.  A population size below 50, for example, is bad; a population between 50 and 100 is good; and, a population over 100 is bad.

 

Stephens et al. (1999) continue with their definition to explore the distinction between the component Allee effect and the demographic Allee effect.  The distinction lies basically in the scale, where the component Allee effect is the positive relationship between any measurable component of individual fitness (breeding, feeding, or defense) and population density; the demographic Allee effect is the positive relationship between the overall individual fitness and population density.  So adding component Allee effects together produces an overall demographic Allee effect.  This is where a lot of the fun equations come into existence, but I’m going to avoid all that because I don’t want to get away from the whole point of this post, which is: how does the Allee effect apply to my project.

 

Black abalone on San Nicolas Island (B. Blaud)

Black abalone on San Nicolas Island (B. Blaud)

This leads back to the basic core question I’m trying to answer dealing with recovering black abalone populations on San Nicolas Island.  How close together do two black abalone need to be in order to successfully reproduce?  My question delves into a component Allee effect, just one aspect of individual fitness that’s investigated as a result of population density.  The more dense abalone are, the closer they are located, the more their gametes will mix and more babies will be made.  When 99% of the population was eliminated, numbers drastically decreased and density was also affected.  As they are recovering, numbers are slowly rebounding, but more interesting is the rate the population density is increasing.  Black abalone are clustering closer together as they recover, which leads us to another interesting story about the Nearest Neighbor Data.  But that is a story for another time…

 

Stephens, P.A., W.J. Sutherland, and R.P. Freckleton.  1999.  What is the Allee effect?  Oikos, Vol. 87(1): 185-190.





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…

 





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!





RocketHub Launches my Project

14 11 2012

Black abalone on San Nicolas Island (B. Blaud)

I have definitely over-used this word lately, but in my overwhelming job to be a part of such a fun way to raise money, I can’t think of any more appropriate vocabulary, so I will keep using the word.  I am EXCITED!!!  SciFund has helped me become organized in the crowfunding world, and through RocketHub, I have launched a fundraising campaign, the Black Abalone Dating Service.  My goal is to raise $2500, and to help people understand what I’m trying to accomplish and what the money will be used for, I put together a short description and video.

 

Black Abalone Dating Service

Are you a lonely black abalone, seeking a nearby abalone of the opposite sex to spawn near and reproduce with?  If that’s the case, you’re not alone!

There were once so many black abalones in California that this intertidal sea snail was stacked 6-deep in the open, so finding a mate wasn’t hard.  After a disease culled them to a fraction of their original numbers, they are now more cryptic and located much further from potential mates.

Black abalones are broadcast spawners, meaning they release their eggs or sperm directly into the water in a Hail Mary attempt to reproduce, hoping they are close enough for their gametes to mix and the eggs to fertilize.  But the question is: how close is close enough?

 Fundraising Target: $2,500

Because black abalones are an endangered species and we have not been able to successfully spawn them in laboratory settings, we use small particles as surrogates for black abalone sperm and eggs in our experiments.  We release these particles in the intertidal and collect water samples to measure the concentration of sperm and eggs over distance and time.  The cost of the particles for each experimental run is approximately $2,500.  We aim to raise enough to fund one run in the hopes of finding out how close two abalones have to be in order to successfully reproduce.   Any money we receive over our fundraising target will go to funding further experimental runs to see how different variables affect the necessary proximity two abalone must be in to successfully reproduce.

 

You can view the video through the SciFund link at: http://www.rockethub.com/projects/11912-black-abalone-dating-service

 

I literally check the website 10 times a day to see if anyone has contributed, and to show that I believe in myself and my project, I was the first to fuel it with a donation.  Any help is appreciated, seriously appreciated.  So go check out the link, and open your wallets!





Results, Revisions, and Revamping

7 11 2012

Recap: So I set up the experiments, collected the samples, counted all of the different sized particles, and entered them into a spreadsheet.  Now what?

An example of R code I used to generate a figure (B. Blaud)

 

I have a secret… I hate R.  Ok, I don’t mean that; I just don’t like it as much as I should, as a scientist.  R  is an amazing free data analysis tool that you can do anything on, as long as you speak computer (specifically, S Language).  I, unfortunately, don’t.  Learning to speak computer is like learning a whole new language, which takes time, dedication, and motivation to practice.  To learn a language, you sing the alphabet song, study vocabulary on flashcards, and practice grammar by writing meaningless sentences in four different tenses over and over.  Learning a computer language is similar, and so I cheat.  I use Excel.  Instead of fourteen lines of code that may be necessary to generate a figure of the average volume of particles in each run, I just hi-lite the desired cells, and type “=AVERAGE(B2:B115)” to compute my run average, and fill in the blanks under the “Charts” tab to make a pretty picture.

 

Why is this a dirty secret?  Several great blogs and articles out there explain in great detail the difference between the two types of software, mainly focusing on user-friendliness and statistical power.  Overall, though, in the professional world, it’s frowned upon to use something like Excel.  So for now, in a very public blog that I share with everyone I know and a lot of people I don’t know, I will just ask that you keep my secret until I become more proficient in the more professional software – which I am working on, slowly but surely. 

This is from my PowerPoint presentation on my project showing figures generated on Excel.  The legend in the top right shows how the colors of the bars change with the increase in probability of successful fertilization in sperm concentration. (B. Blaud)

Now, I get back to the actual results.  It was incredibly exciting counting egg sized particles.  I was finding them in samples as far as 8m away from where they were released, yet they were still hanging out at each distance as long as 5 minutes after the initial release.  Pretty cool!  This means that in the turbulent intertidal, where water is constantly coming and going, the eggs still remain in the immediate area, but just disperse more depending on how the strength of the water activity.

Abalone egg
(oceanlink.info/Conservation/
abalone/BHCAP/
development.html)

 

The sperm sized particles were what I was initially more invested in, seeing as how it’s the guys that really get the girls going, and it’s the concentration of sperm that determine fertilization success.  You see, for an egg to be fertilized, there just needs to be at least one egg present, but it requires 10,000 to 1,000,000 sperm per milliliter (mL) of seawater for a 60 to 100% probability of fertilization (according to laboratory studies).  Fertilization can occur with as few as 100 to 1,000 sperm/mL of seawater, but the probability drops below 10%.  In my results, the concentrations of sperm rarely went above 500 sperm/mL, and only twice were above 1,000 sperm/mL.

 

At first, I was disheartened by these results, thinking this means that my experiment failed or spawning success was extremely unlikely in the intertidal.  After putting this into more realistic terms, however, I realized that my experiment only represents a single male spawning for a period of 10-minutes, whereas actual spawning events may take as long as an hour, and gametes continue to remain viable (alive and able to fertilize) for an additional hour after being spawned.

 

So I revised my project.  Instead of a single male spawning for 10 minutes, I’m going to compare results from a single male spawning for 30 minutes (with samples collected 30 minutes after spawning commences) with three simultaneous males spawning for 10 minutes, and see how those results differ from my original experimental runs.  And in the future, I will avoid coulter counters at all costs!!!  (That’s the revamp).