My Days with the Shrimp

Deanna Hausman, The University of Texas at Austin

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In “What can baby shrimp teach us about oil spills,” I discussed the problem of UV enhanced toxicity of oil. In other words, the fact that UV rays can cause molecules in oil known as PAHs to become more harmful than they would be otherwise. I also discussed the fact that this summer, I will be studying the effects of oil toxicity on grass shrimp, or Palaemontes pugio, an important estuarine species that cycles nutrients through the food chain. Because oil spills are always complex, and organisms can be exposed to oil in many different ways, from the sediment they walk on to the water they swim through, a variety of experiments are needed to get a better understanding of this issue.

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A few of the many PAHs- the compounds in oil that harm marine life Photo from: http://www.crawfordscientific.com/newsletter-2008-12-dedicated-HPLC-GC-columns-PAH-analysis.htm

The first and simplest of the experiments I conducted was the developmental test. In this test, I basically mixed oil and seawater in a giant blender, then took out the water with the oil dissolved in it. Then, I made several dilutions, creating several concentrations of the oily water. Then, I took 6-well plates and filled them up, and placed a single, 24-hour old shrimp in each well. Then, I put these plates in an incubator under UV and non-UV light, and waited for 4 days. After that, I moved the shrimp into clean water, counted how many died, and am currently monitoring them to see how the oil exposure in early life impacts their ability to grow into healthy juveniles.

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Shrimp being monitored after initial oil exposure

Another experiment I conducted essentially followed the same procedure as above, but instead of watching them as they grew, I analyzed the shrimp after their 96-hour oil exposure to see whether the oil affected the concentration of a hormone, known as an ecdysteroid, that controls their molting. Essentially, if the concentrations of this steroid are off in a shrimp it can’t grow properly, so it’s very important!

I’m also conducting an oil sheen test. In this experiment, I place 40 larval shrimp in an aquarium, some caged on the bottom and some swimming freely, and then place an extremely thin oil sheen on top. One aquarium goes under UV light and the other goes under fluorescent light, and after exposure I analyze whether the sheens have had a harmful effect. Whether thin oil sheens are toxic is something that’s not very well understood in this species, so it will be very interesting to see the results.

Finally, I’m conducting an experiment to see what occurs when oil is mixed in with sediment. Essentially, this involves putting sediment from an estuary in a jar, adding oil, and tumbling it around so that the oil is completely mixed in. Then, the sediment is placed into beakers along with water and 24-hour old shrimp, and put under UV and non-UV light for 24 hours, in order to see what mortality occurs. This will perhaps be the most informative experiment, as grass shrimp spend most of their time on the seafloor, so if they’re going to be exposed to oil, it will likely be from the sediment they’re walking on.

In short, I have my hands full this summer! It will be very interesting to see the results. Hopefully, this will increase our knowledge of the harmful impacts oil spills can have to estuarine organisms, and allow NOAA and oil spill analysts to make better predictions of the long-range impacts of oil spills. Ultimately, this may help them make better clean-up decisions.

Thank you to my mentors, Dr. Marie Delorenzo and Dr. Paul Pennington, for their guidance. I’d also like to thank Katy Chung for all her help and expertise. This research is funded through the National Science Foundation.

Some Dramatic Microorganisms and Targeted Genetic Analysis

Emily Spiegel, Bryn Mawr College

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Genetic analysis has become the name of the game in many fields of biological research. Genes encode proteins, and in biology, proteins are king. Proteins guide biological pathways throughout the entire organism, so if you can track the genes, you can understand how the animal functions. Advances in technology like CRISPR, RNA sequencing, and PCR have improved the accessibility and accuracy of high-level genetic analysis in laboratories across the world. Some scientists utilize this technology to study the entire genome of an organism, while others attempt to understand the response of specific genes to various environmental factors or other external influences. This summer, I’m conducting an experiment focused on the latter. I’ll be studying how the polar algae species, Fragilariopsis cylindrus (affectionately known as Frag) copes with environmental stress by reproducing sexually. To do so, I’ll use targeted RNA sequencing to track genes related to sexual reproduction.

In order to understand how a Frag, responds to environmental stresses, you need a lot of algae. I reared nearly 100 liters of this algae in different artificial conditions. These conditions varied by two factors: photoperiod (the length of day and night), and nutrient levels. If you missed my previous post, “Stressing Out My Algae,” you should check it out for more details on the background for this experiment. We suspect that in conditions of stressful light energy (24 hours of continuous light), Frag will respond by reproducing sexually as opposed to its normal asexual mode of reproduction. This could possibly be a mechanism to rid itself of excess energy in times of stress, since sexual reproduction is more energetically expensive than asexual reproduction. By reproducing sexually, Frag may improve its chances of survival against this stress. Compounded with this is our hypothesis on nutrient deprivation. Previous experiments have shown that when a major nutrient, nitrogen, is limited, the algae cannot grow at full capacity and sexual reproduction is inhibited. We predict that when the stress of nitrogen limitation is combined with the stress of high light energy, we’ll see a reduction in the algae’s ability to survive in the stressful conditions due to the inhibition of sexual reproduction. So if we stress out the Frag enough and take away their ability to have sex, they’ll probably die. They’re some very dramatic microorganisms.

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24 bottles of algae were grown in six different experimental conditions varied by length of light exposure and nutrient levels. Algae was reared in 4-liter bottles filled with seawater.

So we grew our Frag, four bottles per six experimental conditions. Every day for eight days we extracted biomass from the bottle. From this sample we could test chlorophyll levels and cell counts, both of which give us a good idea of how well the algae in that bottle are growing in their conditions. We also took samples to be used for RNA extraction. Remember how genes encode proteins and proteins are king? Well before you can have your protein product, you need RNA. You’ve probably heard of DNA, which is the double stranded genetic cookbook. RNA is its single stranded offspring, which is then used as a the direct template to make proteins. A lot of genetic analysis therefore looks at RNA instead of DNA in order to understand how genes are being transcribed for protein production. We’re currently working on extracting the RNA from the original biomass sample and then running that pure RNA through a specialized machine called Nanostring. This is extremely targeted analysis, as Nanostring focuses in on the specific RNA we’re most interested in. In this case, we’re interested in RNA which is encoded from genes related to sexual reproduction. Using Nanostring will tell us how active the genes for sexual reproduction are in each bottle, which we can analyze to derive any correlation between our environmental stress factors and sexual reproduction.

If our hypothesis is correct, then we’ll see the greatest expression of sexual reproduction genes in the conditions of high light energy (24 hours of continuous light). We’d expect to also see low growth performance in nitrogen limited populations, indicated by low cell counts and chlorophyll levels. In these populations we predict we’ll see little if any expression of genes related to sexual reproduction. By the end, we’ll hopefully have a clearer picture of how phytoplankton like Frag deal with environmental stress.

Funding for this project is provided by the National Science Foundation in collaboration with the College of Charleston Grice Marine Laboratory and the National Oceanic and Atmospheric Administration. Acknowledgements to the entire lab of Dr. Ditullio and Dr. Lee in the Hollings Marine Laboratory facility.

Our complicated relationship with chemicals

Nina Sarmiento, Binghamton University

Chemicals found all around us that have been altered, mimicked, and synthesized to be added to our products, are behind the success of our modern society. They have made our plastics strong, our crops prosperous, and our medicines effective. But I have always wondered about the toxicity of these chemicals.  When you look at their biological activity, a chemical might possess the potential to do harm, like interfere with biological processes. The safety of a potentially harmful chemical is based on exposure and dose. It is important to know if we are touching it, eating it, or breathing it in, and for what period of time. The study of evaluating the harmful effects of substances on exposed organisms is what toxicology is all about. They have such an important job because their findings influence what we know is safe and unsafe, for us and organisms all around us.

I learned early on from pursuing biology that we are exposed to many things we are unaware of. Not only are we exposed to potentially harmful chemicals, but we facilitate exposure to other living organisms that may more sensitive. Take dogs for simple example. The toxic dose of something like chocolate for humans is very high, whereas leaving a small amount of chocolate out for a dog to eat could easily kill it.  Rachel Carson is someone I greatly admire, whose work on the pesticide DDT also exemplifies this reality. Food crops were the target for DDT, but birds were indirectly ingesting it, explaining the decline in the Bald Eagle population.  She is one of the people that sparked my interest in ecotoxicology, looking at the effects of harmful substances on ecology, not just humans.

unnamedThis is an example of some of the questions ecotoxicologists ask when there is a potentially harmful substance found in the environment. Photo credit: globe.setac.org.

Here is a chemical product you may not suspect as a threat, sunscreen. In sunscreens, UV filters protect you from getting burned, but also can act as endocrine disruptors, altering hormones and growth (1). Sunscreens are only meant for human skin, however they end up in our lakes, rivers and oceans through swimming or through waste water treatment effluent (2). Unintentionally, many more organisms become exposed.

Toxic-Sunscreen

Photo credit: thesleuthjournal.com

In my project I will be using sea urchins as a model organism to study the effects sunscreens might be having on coral reefs.  I am learning how to preform toxicity tests on sea urchin sperm and embryos which involve an exposure period with sunscreen formulations and then evaluation of effects. I hope to investigate if the chemicals from sunscreens in the water can have negative impacts on coral reproduction.  My work can potentially help create understanding of how humans are contributing to coral reef decline, and influence others to take action to protect them.

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This is a picture of sunscreen water accomodated fractions (WAFs) I am making. They are a mix of sunscreen and seawater and I will be exposing the sea urchin embryos to each solution!

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This is me in the lab with a microscope I use to look at sea urchin sperm and embryos! Photo by Bob Podolsky

My research is funded by the National Science Foundation and College of Charleston partnered with National Oceanic and Atmospheric Administration

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Works cited:

1 Krause M.,, Klit A., Jensen M., Soeborg T., Fredrickson H., Schlumpf M., Litchensteiger W., Skakkebaek N E., Drzewieck K T. 2012. Sunscreens: are they beneficial for health? An overview of endocrine disrupting properties of UV-filters. International journal of andrology. 35 424-436.

2Kyungho C., Kim  S. 2014. Occurances, toxicities, and ecological risks of benzophenone-3, a common component of organic sunscreen products: a mini review. Environment International. 70 143-157.

The real beauty of coral reefs

Nina Sarmiento, Binghamton University

The beauty of a coral reef is undeniable. Over four thousand species of fish, 800 species of coral, invertebrates, and large macro fauna coming together in one place is sure to create a thrilling visual experience. You might be surprised to learn that these remarkable places filled with twenty five percent of marine life, constitute less than one percent of the ocean floor.1 But you don’t have to be lucky enough to travel to a coral reef to fully appreciate its beauty. The real value of reefs comes from their unsuspecting roles in sustaining life as we know it.

photo cred: fmap.ca

photo cred: fmap.ca

Fish from approximately half of our global fisheries, at one point spent a part of their life in coral reefs.2 The unique habitat hard corals provide is perfect for spawning and juvenile life for many species, which may later end up in other parts of the ocean. Fishermen make their livelihood from these reefs, harvesting an average of fifteen tons of seafood annually per square kilometer.3

As for people living on our tropical coastlines, reefs play a crucial role in protecting life on land. It is in the beauty of the long braches of Copra palmata, among other corals, that dangerous storms and waves are softened. Corals roughness and their shallow locale dissipate wave energy, and we have a natural barrier that safeguards our homes.4

Acropora palmata – “Elkhorn coral” Photo cred: coral.aims.gov.au

Acropora palmata – “Elkhorn coral”
Photo cred: coral.aims.gov.au

The importance and intrigue of coral reefs has led to studying many of the organisms and interactions there, leading to new understandings of many aspects of organism biology and evolution. Additionally research has uncovered new medicine from extracting compounds unique species have, giving reefs an importance in future medical interests.

The paradox is that, of all the reasons why we appreciate coral reefs, it is we, the human species that are not having a good effect on them. In fact we are seeing reef decline in many parts of the world because of our actions.5

This summer I am delving into studying one of the possible reasons for this decline; a chemical threat to coral that may not be obvious at first, but could have significant implications on their ability to survive and reproduce.

Stay tuned to hear about my project and the amazing opportunity I have to be a part of the effort to preserve these beautiful communities.

References:

1 Spalding MD, Ravilious C, Green EP. 2001. United Nations Environment Programme, World Conservation Monitoring Centre. World Atlas of Coral Reefs. University California Press: Berkley. 416.

2 US Coral Reef Task force. 2000. The National Action Plan to Conserve Coral Reefs. Washington DC: US Environmental Protection Agency. 34.

3 Ceasr H. 1996. Economic Analysis of Indonesian Coral Reefs. Washington DC: The World Bank.

4 Lowe JR, Falter JL, Bandet MD, Pawlak G, Atkinson MJ, Momismith SG, Koseff JR. 2005. Spectral wave dissipation over a barrier reef. Journal of Geophysical research. 10: C04001.

5 Nystrom M, Folke C, Moberg F. 2015. Coral reef disturbance and resilience in a human-dominated environment.

Funding for my research comes from the National Science Foundation in partner with The College of Charleston and the National Oceanic and Atmospheric Administration

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