Living Life as a Sea Urchin Momma

Hailey Conrad, Rutgers University

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Me working hard to make my sea urchin babies

For my project I am using the same technique that the father of genetics, Gregor Mendel, used to establish his Laws of Heredity: cross breeding. So, I have to breed and raise a whole lot of sea urchins. For a refresher, I’m trying to determine if there is heritable genetic variation in how sea urchin (specifically an Arbacia punctulata population from Woods Hole, Massachusetts) larvae respond to ocean acidification. To do this, I’m rearing sea urchin larvae in low and high carbon dioxide conditions and measuring their skeletal growth. I’m breeding 3 sea urchin males with 3 sea urchin females at a time, for a total of 9 crosses. To tease apart the impact of genetic variation on just the larvae themselves, I will be fertilizing the sea urchin eggs in water aerated with either current atmospheric levels of carbon dioxide, about 410 parts per million, or 2.5 times current atmospheric carbon dioxide levels, about 1,023 parts per million. Then, I will be rearing the larvae in water aerated with either 409 ppm CO2 or 1,023 ppm CO2. This will give me four different treatments for each cross, giving me 36 samples in total. By fertilizing and rearing them in the same and different levels of carbon dioxide I will be able to see how much of an impact being fertilized in water with a higher carbon dioxide concentration has on larval growth versus just the larval growth itself. It’s important for me to make that distinction because I just want to identify genetic variation in larval skeletal growth, and separate out any extraneous “noise” clouding out the data. I’m rearing the larvae in a larval rearing apparatus. Each of the 36 samples will be placed in jar with water aerated with the correct CO2 treatment. Each jar will constantly have atmosphere with the correct CO2 concentration bubbled in. Each has a paddle in it that is hooked to a suspended frame that is swayed by a motor. This keeps the larvae suspended in the water column. The jars are chilled to 14 C by a water bath.

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My larval rearing apparatus

After a 6-day period the larvae are removed from the jars and their skeletal growth is measured. They are preserved with 23% methanol and seawater and frozen.

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An Arbacia punctulata pluteus

You’re probably curious how the heck I am able to measure the larva’s skeletons. They’re microscopic! Well, I use a microscope coupled to a rotary encoder with a digitizing pad and a camera lucida. Which, looks like this:

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A microscope coupled to a rotary encoder with a digitizing pad and a camera lucid hooked up to a computer

This complicated-sounding hodge-podge of different devices enables me to do something incredible. I can look through the microscope at the larva, and also see the digitizing pad next to the microscope, where I hold a stylus in my hand. When I tap the pad with the stylus and the coordinates of various points on the anatomy of the plutei that I am tapping at get instantly recorded on my computer! The rotary encoder is the piece attached to the left side of the microscope and it enables me to record coordinates in three dimensions. Then, I can use those coordinates to calculate the overall size of the skeleton. My favorite part of doing science is learning how scientists are able to do the seemingly impossible- like measuring something microscopic.

After I gather all of my data, I will do some statistical analysis to see the affect that the male parents have on the skeletal growth of their offspring. I will not be focusing on the impact that females have on the skeletal growth of their offspring. The quality of the egg itself could be an influencing factor on the size of the offspring, whereas sperm is purely genetic material. Like how I’m trying to isolate the influence of ocean acidification during larval rearing from during the act of fertilization, I am trying to isolate just genetic influences on larval skeletal growth from egg quality. Check back to see how it goes!

Special thanks to the National Science Foundation for funding this REU program, the College of Charleston and Grice Marine Laboratory for hosting me, and Dr. Bob Podolsky for mentoring me!

 

 

 

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Playing with Plutei

Hailey Conrad, Rutgers University

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Me! Photo Credit: Kady Palmer

Ocean acidification is known as climate change’s evil twin. When the pH of ocean water drops, carbonate ions in the water form carbonic acid instead of calcium carbonate. Calcium carbonate is the form of calcium that marine animals that have calcium-based skeletons (like us!) and shells use to build their bones and shells. Having smaller and weaker skeletons or shells impacts their ability to survive. However, some individuals within certain species or populations of species have genes that make them more resistant to ocean acidification. If these individuals are able to pass on these genes to their offspring, then the species has the ability to evolve in response to ocean acidification instead of going extinct. This summer I’m working with Dr. Bob Podolsky in College of Charleston’s Grice Marine Field Station to study the extent to which ocean acidification affects Atlantic purple sea urchins, Arbacia punctulata. We are specifically trying to see if any individuals within a population from Woods Hole, Massachusetts, have any heritable genetic resistance to the negative impacts of ocean acidification. We hypothesize that there will be genetic resistance given that the northern Atlantic coast naturally has lower levels of saturated calcium carbonate, so a population that has evolved to live in that type of environment should have some resistance to lower calcium carbonate levels already (Wang et al 2013). We’re using a basic cross breeding technique to rear Arbacia punctulata larvae to their plutei stage, when they have four main body rods. At this stage they look less like sea urchins than they do like Sputnik!

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A sea urchin pluteus larvae with four body rods

Then, we will look to see if any of the male parents consistently produce male offspring that are more resistant to ocean acidification.  If males like these exist within this population, then the species has the capacity to evolve in response to ocean acidification, instead of going extinct! This is a very big deal, and could potentially be very hopeful. Even if we don’t get the results that we are hoping for, the results of this research could inform policy and management decisions.

Literature Cited:

Wang, Z. A., Wanninkhof, R., Cai, W., Byrne, R. H., Hu, X., Peng, T., & Huang, W. (2013). The marine inorganic carbon system along the Gulf of Mexico and Atlantic coasts of the United States: Insights from a transregional coastal carbon study. Limnology and Oceanography, 58(1), 325-342. doi:10.4319/lo.2013.58.1.0325

Thank you to the National Science Foundation and College of Charleston’s Grice Marine Laboratory for funding my project. And, special thanks to Dr. Bob Podolsky for being a wonderful and supportive mentor!

 

Exploring the “Secret Garden”

Christine Hart, Clemson University

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On a walk along the beach, have you ever noticed the garden growing at the water’s edge? During low tide patches of green and gold speckle the sand, growing what researchers have called a “Secret Garden.”

The “Secret Garden” is made up of a variety of microorganisms like cyanobacteria, flagellates, and diatoms. These small, sand-dwelling organisms are collectively known as benthic microalgae (BMA). BMA are responsible for 50% of primary production in estuary systems through photosynthesis and an extracellular polymeric secretion. Though small, these photosynthetic powerhouses form the basis of ocean food webs. BMA are also important indicators of ecosystem health. Scientists have documented the response of BMA to a variety of environmental conditions. As humans change natural estuary conditions, BMA will serve as a bioindicator for changes in ecosystem health.

The visible patches of green and gold at low tide indicate an increasing density—or biomass—of BMA. Currently, researchers do not know the mechanism for the visible change in BMA biomass. Our study will focus on two possible mechanisms of biomass change. One mechanism may be the vertical migration of BMA to the top of the sand.  The increase in biomass could also result from growth among BMA species due to sunlight exposure.

In addition to the unknown mechanism, the particular BMA species associated with the green and gold sheen have not been well studied. Like plants in a garden, BMA species are diverse and serve their own roles in maintaining a healthy environment. To better use BMA as a bioindicator, we will characterize the type of BMA contributing to the visible biomass changes.

Our study will focus on the mechanism of changes in biomass during low tide, while also identifying changes in the presence of BMA species. The results from the study will give us a greater understanding of the BMA that are essential to estuary systems. This information will establish a basis of BMA dynamics that can be used as an indicator of the health of estuaries.

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Thank you to my mentor, Dr. Craig Plante, and my co-advisor, Kristina Hill-Spanik, for their support and guidance.  This project is funded through the National Science Foundation, and supported by The College of Charleston’s Grice Marine Laboratory.

 

Literature Cited

Lobo, E. A., Heinrich, C. G., Schuch, M., Wetzel, C. E., & Ector, L. (n.d.). Diatoms as Bioindicators in Rivers. In River Algae (pp. 245-271). Springer International Publishing. doi:10.1007/978-3-319-31984-.

MacIntyre, H.L., R.J. Geider, and D.C. Miller. 1996. Microphytobenthos: the ecological role of
 the “Secret Garden” of unvegetated, shallow-water marine habitats. I. Distribution, abundance and primary production. Estuaries 19: 186-201.

Plante, C.J., E. Frank, and P. Roth. 2011. Interacting effects of deposit feeding and tidal resuspension on benthic microalgal community structure and spatial patterns. Marine Ecology Progress Series 440: 53-65.

Rivera-Garcia, L.G., Hill-Spanik, K.M., Berthrong, S.T., and Plante, C. J. Tidal Stage Changes in Structure and Diversity of Intertidal Benthic Diatom Assemblages: A Case Study from Two Contrasting Charleston Harbor Flats. Estuaries and Coasts. In Review.

Underwood, G.J.C., and J. Kromkamp. 1999. Primary production by phytoplankton and 
microphytobenthos in estuaries. Advances in Ecological Research 29: 93-153.

 

Hook, Line and Sinker

Sierra Duca, Goucher College

Spotted seatrout, Cynoscion nebulosus, are important recreational fish that range from the Atlantic coast to the Gulf of Mexico. They are also good indicators of environmental changes in estuarine habitats since all of their life stages are found in estuaries1. To reiterate, I am studying muscle softness in spotted seatrout induced by the parasite Kudoa inornata. Several Kudoa species are notorious for causing this muscle softening, which makes the meat of the fish go bad faster than in uninfected fish2. This is an issue with fish that are consumed by people, such as the commercially important farmed Atlantic salmon2. seatrout for blogg

Fig 1. Spotted seatrout that were caught via trammel netting (PC: Sierra Duca).

First of all, to study this I need fish. While the mode of infection of Kudoa parasites is not well understood, it is presumed that wild spotted seatrout have a higher rate of infection of Kudoa inornata; therefore, I needed some wild spotted seatrout. In addition to the traditional hook and line approach of fishing for spotted seatrout, I was able to join a group at the South Carolina Department of Natural Resources (SCDNR) as they went trammel netting. In comparison to other nets, trammel nets have three layers of netting that vary in size in order to catch fish of various sizes. trammel net for blog

Fig 2. This illustration depicts the basic function and structure of a trammel net. A similar such device is used by the SCDNR to catalogue fish in specific sites over time in order to study the changing population dynamics of various fish species.

Once I have the fish I fillet, refrigerate, and take muscle biopsies at time points between 0-6 days, which is the most likely time that the fish would be consumed. I test the firmness of these muscle biopsies, as well as the parasite density. What I am trying to accomplish is to establish whether or not there is a link between parasite density and accelerated muscle softness (which causes the meat to go bad faster in infected fish), and if the rate of muscle softening changes over the course of 6 days. Ultimately the project will help increase our understanding of the effects of Kudoa inornata on the muscle of spotted seatrout. plasmodia for blog

Fig 3. This image (under 100x magnification) displays a plasmodium structure that contains a cluster of spores (known as myxospores) of Kudoa inornata in the muscle tissue of spotted seatrout. One way that I quantify parasite density is by looking at the average area of plasmodia. I can do this because generally larger plasmodia are found in the more infected fish  (PC: Sierra Duca).

Literature Cited 1Bortone SA (ed) 2003: Biology of the Spotted Seatrout. CRC Press. Boca Raton, FL, 328 pp 2Henning SS, Hoffman LC, Manley M (2013) A review of Kudoa-induced myoliquefaction of marine fish species in South Africa and other countries. S Afr J Sci. 109: 1-5

Photo Source (Fig 2): http://thewikibible.pbworks.com/w/page/22174694/Fishing%20in%20the %2Bible%20and%20the%20Ancient%20Near%20East

Acknowledgments The Fort Johnson REU Program is funded by the National Science Foundation. This research is made possible through the mentorship of Dr. Eric McElroy and Dr. Isaure de Buron.  In addition, I would like to thank the College of Charleston and the South Carolina Department of Natural Resources for providing the help and facilities necessary for my project.

New Philadelphia to Charleston

Aaron Baumgardner, The University of Akron

Coming from the landlocked small town of New Philadelphia in the Midwest, I feel like I’m dreaming when I realize I’m spending my summer researching in Charleston, SC. I’m thankful for the opportunity that my mentor, the College of Charleston, and the National Science Foundation has given me to learn and grow in my scientific ability.

However, I do not believe I would be where I am today if it weren’t for my Aunt Jane. She is the only member of my family with a background in science, and even though she is hundreds of miles away at UPenn, she is always an email or phone call away. She has always shown an interest in my academics and will always be there for any advice I may ask. She has helped me develop my professionalism and offered insight on which graduate schools are worth going to.   Because of her, I can finally realize it’s not a dream. It’s reality that I’m spending my summer in Charleston, SC. It’s because I’ve worked hard in school and reached out for opportunities for me to mature as scientist. And I owe her so much for pushing me to succeed.

Thank you Aunt Jane!

Parasitic Kudoa inornata causes muscle deterioration in spotted seatrout

Sierra Duca, Goucher College

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This summer I am working at the South Carolina Department of Natural Resources at Fort Johnson under the mentorship of Dr. McElroy and Dr. Isaure de Buron. Being part of an REU program, I am looking forward to gaining some research experience and learning more about career possibilities in the biological sciences. In terms of my research, I am researching the effects of a microscopic parasite, Kudoa inornata, on spotted sea trout (see image below).

Seatrout

Photo source: http://www.kayaking-north-america.com

There are many Kudoa species that infect host fish worldwide. Several of the Kudoa parasites have spores (known as myxospores; see image below) that proliferate inside of the muscle fibers of the host fish (Harrel et al. 1985). In some cases the  parasites wreak havoc on the quality of the meat after the fish dies. For example, infected fish may have unsightly cysts or decreased meat quality (Moran et al.), both of which are unappealing to consumers. Don’t worry, the majority of Kudoa infected fish are not directly harmful if consumed by humans; however, if the deterioration of the muscle tissue is accelerated, like any meat, the quality will decrease sooner as compared to uninfected fish. This process of muscle deterioration is what I am studying with K. inornata infected spotted seatrout. I am looking at the rates of this deterioration during various time frames, from 0 to 6 days after the fish dies, in order to discern if there is a relationship between parasite presence in spotted seatrout and muscle softness.  Ultimately, this research can be used to better understand the biology of K. inornata and to determine the best time frame to consume infected spotted seatrout.

picc     Individual spores of Kudoa inornata (Photo source: Dyková et al., 2009).

Literature Cited

Dyková I, de Buron I, Fiala I, Roumillat WA (2009) Kudoa inornata sp. n. (Myxosporea: Multivalvulida) from the skeletal muscles of Cynoscion nebulosus (Teleostei: Sciaenidae). Folia Parasitology 56: 91-98

Harrel LW, Scott TM (1985) Kudoa thyrsitis (Gilchrist) (Myxosporea: Multivalvulida) in Atlantic salmon, Salmo salar L. Journal of Fish Diseases 8: 329-332

Moran JDW, Whitaker DJ, Kent ML (1999) A review of the myxosporean genus Kudoa Meglitsch, 1947, and its impact on the international aquaculture industry and commercial fisheries. Aquaculture 172: 163-196

Acknowledgments

The Fort Johnson REU Program is funded by the National Science Foundation. This research is made possible through the mentorship of Dr. Eric McElroy and Dr. Isaure de Buron.  In addition, I would like to thank the College of Charleston and the South Carolina Department of Natural Resources for providing the help and facilities necessary for my project.

Shrimp kabob, shrimp gumbo…shrimp sickness?

 

Alessandra Jimenez, Whitworth University

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Are you a fan of shrimp? You’re not the only one – billions of people around the world depend on shrimp fisheries and aquaculture for this wonderful source of food. Other predators in the sea rely on shrimp for their daily meals. Here’s the catch: shrimp may not last long enough to make it to your plate. Like us and other animals, crustaceans in general have to deal with so many obstacles that threaten their survival. One obstacle that is not often thought about is bacterial infection. Did you know that seawater is literally teeming with hundreds of millions of bacteria? The only way a shrimp can make it is by using its immune response – the “quick, potent, and effective” way of defending against a huge, microscopic army! Sounds like the perfect shield, right?

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Shrimp is a common food source for many people. @Leslie Fink

Turns out that, like everything else in the science world, immunity comes at a big cost. It has been recently discovered that the immune response in crabs and shrimp against bacteria actually has a bad side effect: metabolic depression. In fact, the way the shrimp gets rid of bacteria in its bloodstream is by moving the bacteria to the gills, where it gets lodged and stays there for quite some time. The consequence? The lodged bacteria block blood flow through the gills, and the shrimp can’t get enough oxygen from the water. (Want to learn more? Click here)

Ouch, talk about a double whammy – fighting sickness plus oxygen blockage. One basic question comes to mind: can the shrimp still do what it needs to do while under such metabolic stress? This is where I come in. This summer, I am working under Dr. Karen Burnett in Hollings Marine Laboratory as an intern through the Research Experience for Undergraduates (REU) program in marine biology. We will be testing whether or not a shrimp’s immune response to a common bacteria affects its ability to perform daily activities. The activity of interest is called ‘tail-flipping’ (fancy name: caridoid escape reaction. Want to learn more? Click here)This really fast, reflex-like action needs to be in top shape for the shrimp to survive from predator attacks and to help it during feeding time.

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Also known as the ‘tail-flip’ reaction, this response is a shrimp’s primary means of escape. @Uwe Kils

The shrimp species of interest is Farfantepenaeus aztecus, or ‘Atlantic brown shrimp’. This fella is a familiar catch for fishermen throughout the Southeastern US and the Gulf of Mexico. This is the first time that a study like this is going to be done on a wild shrimp species in general, let alone this specific type!

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Farfantepenaeus aztecus, aka ‘Atlantic brown shrimp’. @Virginia Living Museum

So, can an immune response impact tail-flipping in wild shrimp? If ‘yes’, would the potentially handicapped shrimp be able to survive in its natural environment? We will soon find out!

Happy shrimping!

Alessandra Jimenez

REFERENCES:

Burnett, L. E., Holman, J. D., Jorgensen, D. D., Ikerd, J. L., & Burnett, K. G. (2006). Immune defense reduces respiratory fitness in Callinectes sapidus, the Atlantic blue crab. Biological Bulletin, 211(1), 50-57.

Fuhrman, J. A. (1999). Marine viruses and their biogeochemical and ecological effects. Nature, 399(6736), 541-548.

Latournerie, J.R., Gonzalez-Mora, I.D., Gomez-Aguirre, S.G., Estrada-Ortega, A.R., & Soto, L.A. (2011).                   Salinity, temperature, and seasonality effects on the metabolic rate of the brown shrimp Farfantepenaeus Aztecus (Ives, 1891) (Decapoda, Penaeidae) from the coastal Gulf of Mexico.Crustaceana 84(12-13), 1547-1560. doi: 10.1163/156854011X605738

Scholnick, D. A., Burnett, K. G., & Burnett, L. E. (2006). Impact of exposure to bacteria on metabolism in the penaeid shrimp Litopenaeus vannamei. Biological Bulletin, 211(1), 44-49.

Many thanks to College of Charleston for hosting my project, Dr. Karen Burnett and Hollings Marine Laboratory for guidance and work space, and NSF for funding the REU program.

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