Vessels, Pumps and VIs – Oh My!

Since my last post I have completely re-designed my chemostat system. Take a look at my last blog post to see a picture of the previous system (aka Cv1.0). Although good in theory (autoclavable culture vessel, large culture volume, inexpensive to implement), many problems came out in the test culture phase.

To begin with, although the culture vessel itself was indeed autoclavable, the bulkhead fittings that connected tubing to the vessel were not. The silicone sealant used to close off gaps was also not autoclavable.

Additionally, the peristaltic pumps for this system quickly became very unreliable and refused to stay synchronized. Since the principle of a chemostat system is dependent on constant, synchronized influx and efflux of media, lack of synchronization in the input/output pumps leads to serious problems such as excessive dilution of the culture or (far worse) draining the culture vessel dry.

The culture vessel for Cv1.0 was also far too large. Topping out at 4 liters, it required an enormous amount of media to keep the culture at steady state.

Because of these problems I have decided to adopt a new system design (Cv2.0). Instead of the previous two pump chemostat system (one input pump and one output pump), the new system is an overflow chemostat. This simply means that the culture vessel has an open port in the side of the flask that drains excess media when the media level rises to the overflow level. This has the great benefit of requiring only one media pump (the input pump) since the overflow port drains media at the same rate that it is being pumped into the system. This new culture vessel is also much smaller (2L media capacity), so media demands should be less. Upgrading to a better quality peristaltic pump seems to have solved the flow inconsistency problems experienced in the previous system.

I have also begun the process of writing a LabVIEW VI (virtual instrument) to control the gas manifold (see picture) in response to culture pH. Slowly but surely, progress is being made.

Gas manifold with solenoid valves and non-compressible gas lines

In between working over my chemostat I have been writing my thesis proposal. It looks like I’ll be defending my proposal sometime in March or April, so I’ve also been working on some preliminary data to relate Alexandrium cellular health to pH. Hopefully I’ll be including this in my next post.

Happy 2013! :)

 

Greetings!

Greetings oceanophiles! As this is my first blog post as a Sea Grant scholar, I feel I should give a little background about myself. I am just beginning my second year as a Ph.D. student with Drs. Tawnya Peterson and Joseph Needoba at Oregon Health & Science University in Beaverton, OR. My degree track is Environmental Science and Engineering, with a focus on Estuary and Ocean Systems. I have always loved the Pacific Northwest, and am constantly amazed that I get to study it for a living.

So what am I actually doing? The goal of my research is to identify links between pH and pCO2 concentration in the water and population dynamics of harmful algal blooms (HABs) in the northern California Current system. My research specifically focuses on the marine dinoflagellate Alexandrium. This little guy is the alga that is primarily responsible for paralytic shellfish poisoning (PSP) events off the west coast of North America. Alexandrium produces saxitoxin, an extremely potent neurotoxin. Shellfish are filter feeders, and accumulate toxins when they feed on HAB species in the surrounding water. When there is an increased number of Alexandrium in the water (as frequently happens in the summer months), saxitoxin builds up to dangerous levels in the shellfish and can cause paralysis in humans and animals. What I want to do is discover whether there is a link between the pH/pCO2 content of the water and population dynamics and toxin production of Alexandrium.

I expect to be spending a lot of time on boats in the next few years, stalking the wild Alexandrium through the Columbia River estuary and out on the coast, but I will probably spend far more time in the lab. To that end, I am in the process of building a chemostat culture system, which will grow algae at a constant rate in a nutritionally static environment. Influx and efflux of media to and from the culture vessel are synchronized to the growth rate of the algae to maintain a constant growth rate.

 

The constant influx of fresh media and efflux of waste will ensure that the nutrient load of the culture vessel remains constant. I also designed the system to automatically monitor and control pH using a custom made pneumatic manifold that will change the pH of the culture vessel by bubbling it with CO2 gas. The monitoring will be accomplished by a Labview program that will also allow for remote monitoring of the system, and send will me alarms if/when something goes wrong. I have already finished the first iteration of the chemostat system and am in the process of working out the bugs (waste overflows, variable pump rates, etc.).

I’m excited to see where this year will take me and to see what new adventures lie in wait, both in the lab and out on the water. Tallyho!

 

 

Spring Transition

This last term I took Comparative Embryology and Larval Biology taught by Dr. Maslakova at OIMB.  In the course we had the rare opportunity to witness first hand the development of many phyla of marine invertebrates.  We also maintained our own cultures of larvae changing their water and feeding them every other day.  The experience of  watching the organisms develop right before our very eyes was one I will never forget.  The class was very informative and I learned a lot about the development of many species.  One of the assignments in the class was to photograph the larva or embryo of an organism and write a brief blurb for the course blog.  I posted about crab zoea, barnacle nauplii and a polychaete called Polygordius.  I highly recommend that folks check out this blog and get a feel for what the larvae of many organisms look like.

 

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In my advisors previous research he found a correlation between the date of the year of the spring transition  and the amount of megalopae returning.  Our hypothesis is that upwelling that occurs following the spring transition brings the megalopae onto the shelf. My goal was to conduct a research cruise the week before the spring transition and a cruise the week after the spring transition to see if the number of megalopae on the shelf increased. Surface plankton samples and CTD casts were conducted on a 10 km transect perpendicular to shore.  We used a manta net (pictured to the left) to collect surface plankton tows.  The net works by skimming across the surface of the water collecting a sample just from the top fraction of water.

IMG_4084We conducted the tows at night and in the surface because previous research has found that the megalopae of Dungeness crabs inhabit the surface of the water column at night.  After deploying the net we towed it behind the boat for 10 minutes (picture to the right).  The cruise before the spring transition was rough and we had wind gusts to 45 knots. The entire crew did great and nobody got sea sick!!! The weather during the cruise the week after the spring transition was much calmer and was rather enjoyable.  We caught many interesting organisms such as salps and doliolids. The night we were out there was a great sunset and we all had the opportunity to watch it.  I am still in the process of analyzing the samples but will post again with the results later.

IMGP0017On April 2nd I put out our light trap to monitor recruitment of megalopae and I have been checking it daily. The photo to the left is of a sample collected on May 28th.  In the photo are approximately 60,000 megalopae that had climbed into the trap during the course of the night.  The recruitment events are highly pulsed and catch has ranged from 60,000 animals in a night to 0 animals in a night.

Orals are done!!!

So last Friday I had my oral exams and I passed!!  Although the last few months of studying have been very stressful, the time I put in was beneficial.   The whole process was much less stressful than I expected but still something I do not want to do again.  I am really looking forward to getting to the microscope and starting to count the samples that I picked up from NOAA.  In addition to counting samples I will be working on designing some rapid deployment mooring for putting out thermistors and an ADCP this summer while I am sampling internal waves.  I look forward to sharing some of the interesting creatures I find in the samples with everyone.

 

Wow where did the term go!?!?

Well the term ended and it was a whirl wind.  The majority of the term was spent either TAing biological oceanography or studying and preparing for my oral exam.  The TAing went extremely well and I really enjoyed getting to hear the lectures again.  Hearing the lectures really helped me to feel more prepared for my upcoming orals.  For part of my project I am using plankton samples collected by NOAA to better understand the vertical migratory patterns of Dungeness crabs.  This quarter I went to pick up the samples from Newport and they filled the back of a pickup truck!!!!  It definitely seems I have my work cut out for me.  However, the majority of the quarter has been spent preparing for orals which has entailed reading countless scientific articles.  Before the oral exam I have to turn in a written form of my proposal so the next few weeks will be spent finalizing this.  I am looking forward to getting past the orals portion of my PhD so I can get to work counting samples and building my computer model.  I will be posting sometime in February to update everyone on how the orals went.

Greetings From the new Malouf Scholar

Hi All,

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My name is Leif Rasmuson and I am graduate student at the University of Oregon’s Oregon Institute of Marine Biology, OIMB.  I am just beginning my second year as a PhD student in Dr. Alan Shanks lab.  I am interested in how hydrodynamics affect the larval stages of commercially important marine invertebrates. For my PhD I am focusing my research on the larvae of the Dungeness crab, Cancer magister.

Dr. Shanks has been using a light trap (see the image below) to capture Dungeness crab megalopae for 10 years.  In Dr. Shanks’ work he has found a strong correlation between the number of returning larvae and the commercial catch four years later.  The number of returning megalopae was found to be negatively correlated with the date of the spring transition, which is an atmospheric shift in winds causing the ocean currents to shift from southerlies to northerlies.  With this change in winds comes a seasonal switch from downwelling favorable conditions to upwelling favorable conditions.  Furthermore, in Dr. Shanks work a negative correlation was found between the number of megalopae recruiting and the Pacific Decadal Oscillation (PDO).  The PDO is decadal oscillation in water temperature from warm to cold temperatures off Alaska caused by variation in the flow of the West Wind Drift as it enters the California Current.  During cool, or negative phase years, more cool water is shifted into the California current causing the southward flowing California current to be stronger.  In negative phase years there were far more megalopae caught in the light trap (about 20 times more), larval return was very strong, suggesting that the increased southward flow may strongly influence the dispersal of the larvae.

 

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For my PhD I intend to use an individual-based couple biophysical computer model to mimic annual hydrographic conditions to examine the correlations talked about earlier. The output from the computer model will be tested against ongoing and historic light trap data to ensure that the model is accurate and provides a realistic description of the movement of larvae.  Since the light trap has been in operation for ten recruitment seasons, I will be able to model many years of ocean currents and PDO regimes allowing me to ground truth the models with historic larval return data ensuring that it is providing an accurate depiction of larval movements.

Since I am just entering into my second year of my PhD I am in the process of preparing for my oral defense that will be occurring this coming fall. Unfortunately this means the majority of my time I will be reading and writing which do not lend themselves to interesting blog posts or pictures. However, I will be sure to post any interesting happenings that occur and fun pictures.