Catch of the Day(s)

Melanie Herrera, University of Maryland College Park

South Carolina is known for its iconic southern cuisine, including a staple of fresh seafood which fuels the buckets of shrimp & grits and “catch of the day”. In order to support this huge industry (and fill the bellies of every South Carolinian), I am conducting an experiment to figure out where this seafood is holing up prior to its demise. Dr. Harold, his graduate student, Mary Ann McBrayer, and I are out on Grice Beach collecting fishes, crabs, shrimps, and much more in order to figure what exactly is there… And what they are using to survive.

Using a seine net, we encircle marine animals in dense and sparse patches of an invasive sea grass, Gracilaria, for collection. We hypothesize that Gracilaria is helping the local economy (a surprising contribution from an invasive species) by creating refuge for young animals. On the beach, we submerge separate samples of animals (from dense versus sparse areas of Gracilaria) into a euthanizing solution to bring them up to the lab for preservation and analysis (Figure 1).

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Figure 1: An example of animals caught in separate habitats at Grice Cove. The left exhibits animals caught in a dense area of Gracilaria and the right exhibits animals caught from a sparse area of Gracialaria. Credit: Melanie Herrera

In the lab, separate samples (dense versus sparse) undergo a few transfers into different fixatives (10% seawater formalin, 25% isopropyl, and 50% isopropyl consecutively) to keep the fish from decaying. After this preserving process, fish and other animals are separated and categorized by family, genus, and species. This categorization enables us to identify and analyze what types of animals and how many of each are using different habitat. Our analysis will give us insight on what type of habitat, either patches dominated by Gracilaria or areas with more open water, benefits fish. Specifically, we will be able to identify if Gracilaria is more advantageous to young fish or if their survivorship is independent from their habitat.

So far, we have collected lots of pipefish, narrow skinny fish that resemble a hair strand-size snake, Atlantic Silversides, a fish that looks exactly like it sounds, and more shrimp than anyone needs (Figure 2). Although some of these animals do not directly contribute to the seafood industry, its presence in the Charleston Harbor can tell us a lot of things. For example, we have seen some fishes that usually stay in warmer waters in the Southern U.S. Their expanding habitat can lead us to some more hypotheses on climate change and warm weather moving northward. In addition, we can find out if Gracilaria has a stake in rearing economically important fish in the future.

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Figure 2: (From left to right) Pipefish, Atlantic Silversides, and Grass Shrimp caught for analysis.Credit: Melanie Herrera

Thank you so much to my mentor Dr. Tony Harold and his lab for his advice and guidance. Thank you to Mary Ann McBrayer for helping me facilitate this project. This research is funded through the National Science Foundation and College of Charleston’s Grice Marine Lab.

 

Expect the Unexpected in Science

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Alessandra Jimenez, Whitworth University

As this internship has recently come to an end, I now begin to reflect on the wonderful yet challenging experience I had conducting observational research on Atlantic brown shrimp (Farfantepenaeus aztecus). In the last few weeks of this 10-week summer program, there was a fascinating yet unexpected turn of events. In particular, results of the experiment pointed to conclusions that I initially found myself unprepared for!

In summary, the focus of this experiment was to test effects of immune response on the ability to escape predators in shrimp. The escape mechanism, called tail-flipping (see video below) is actually powered anaerobically. However, recovery from this energetic behavior absolutely requires oxygen (is aerobic). As further explained in previous blog posts (click here and here), a recently discovered consequence of mounting an immune response against bacterial infection involves depression of aerobic metabolism. So, my mentor and I decided to focus on the recovery aspect (aerobic) of the escape response and predicted that this aerobic process would be impaired in shrimp injected with bacteria. At the same time, we predicted that the anaerobic part of this mechanism would be significantly impacted.

A slow-motion video of an Atlantic brown shrimp juvenile tail-flipping in an experimental tank (c) Alessandra Jimenez

The last few weeks of the internship mainly consisted of analysis, arriving at conclusions, and publicly reporting results. After testing tail-flipping ability (click here for an explanation of how this was tested) in a total of 42 shrimp juveniles, 30 of these were chosen for final analysis. Using a statistics software called Sigmaplot (version 12.5), I conducted tests that basically compared experimental groups based on the two variables I investigated: treatment type (bacteria or saline) and time given after injection (4 or 24 hours). Afterwards, results were deemed important based on significance values assigned by these Sigmaplot tests.

Significant results were very surprising!  Overall, results suggested that metabolic depression (indirectly caused by the immune response) did not have an impact on recovery (aerobic). At the same time, the most unexpected finding of all suggested that bacterial exposure actually increased anaerobic tail-flipping activity in Atlantic brown shrimp juveniles! Thus, this result called for a complete change in focus from the aerobic part to the anaerobic part of this particular escape response.

So, how could I possibly explain the increase in anaerobic processes found through this experiment? After much pondering and going through scientific literature, I formulated a new hypothesis. An important enzyme in crustaceans called arginine kinase is involved in the storage and creation of anaerobic energy that can be used for tail-flipping. Recent studies involved injecting bacteria into live crustacean tissue and comparing arginine kinase expression levels with controls. Results indicated a significant increase in expression in bacteria-injected tissue, especially in abdominal muscle (important for tail-flipping!). Based on these investigations, I now think that there may be a link between immune response and levels of anaerobic metabolism. Further research is required to explore this.

The final stages of the internship included creating and presenting a Powerpoint presentation of our work, and submitting a manuscript of my summer investigation. Overall, this REU internship experience has been challenging yet exciting, and has confirmed my love for marine biological research. As I mentioned at the end of my presentation, “expect the unexpected in science”.

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Picture of me right before giving my Powerpoint presentation (c) 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.

Gruschczyk, B., Kamp, G., 1990. The shift from glycogenolysis to glycogen resynthesis after escape swimming: studies on the abdominal muscle of the shrimp, Crangon crangon. J Comp Physiol B, 753-760.

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.

Yao, C., Ji, P., Kong, P., Wang, Z., Xiang, J., 2009. Arginine kinase from Litopenaeus vannemai: Cloning, expression, and catalytic properties. Fish Shellfish Immunol 26, 553-558.

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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A day in the Shrimp Lab

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Alessandra Jimenez, Whitworth University

Have you ever wondered what it’s like to be a lab researcher who works with live animals? Through this internship, I am experiencing this firsthand in Hollings Marine Laboratory, along with all the responsibilities involved!

A normal workday in the life of a “shrimp intern” is like this: A big part of it is animal care and maintenance. It starts in the morning with a daily visit to the wet lab, where approximately 80 brown shrimp juveniles are kept in four large tanks with circulating water. After feeding them a round of commercial shrimp pellets, I test the salinity of the water in each tank using a refractometer to make sure that each tank has a certain salinity value: 30 parts per thousand, to be exact. I use dechlorinated freshwater and seawater to adjust this value if needed. Besides salinity, I also need to watch out for harmful levels of ammonia (it’s a part of shrimp waste!), nitrates, etc. In usual circumstances, I conduct a water change (replacing old water with new) once a week in order to dilute these chemicals. For the past couple of weeks, however, I have been conducting water changes daily in order to keep ammonia levels neutral in three tanks. Ah, the life of a caretaker of tons of baby shrimp!

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Wet lab. @AlessandraJimenez

Besides animal husbandry, I work on my experiment involving the effects of injection of bacteria on tail flipping (Want to learn more about what I’m doing? click here). I have two shrimp at a time in separate, well-aerated tanks, and they are both from the same treatment group. Shrimp are randomly assigned to one of four treatment groups. These treatment groups are designated according to the treatment type (injection of bacteria or saline) and according to the amount of time between the moment of injection and the tail-flipping procedure (4 or 24 hours). I randomly select two shrimps from the wet lab, weigh them, and keep them in the two experimental tanks overnight so they can get used to the new environment, temperature, etc. The next day, I take each shrimp out of the tank momentarily and quickly inject them with bacteria, or a saline buffer if they are part of the control group. Then, I give them 4 or 24 hours (depending on group type) to rest before conducting the actual tail-flipping experiment. Using a stir-rod (basically, a straight stick), I poke the shrimp lightly to induce tail-flipping, and count how many flips they perform before fatigue. The number of flips here is called ‘initial activity’. Then, I give them 20 minutes to recover in the tank before tail-flipping them again. The number of flips this time is called ‘recovery activity’.

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Experimental tanks @AlessandraJimenez

Why tail-flip them twice? Well, we hypothesize that recovery activity will be impaired in bacteria-injected shrimp versus the controls, while initial activity would probably not be. This is based on how recovery from tail-flipping activities involves aerobic (or oxygen-fueled) metabolism. Since bacteria accumulate in the gills of shrimp and block oxygen uptake (want to learn more? click here), it would make sense that recovery activity would be reduced. Stay tuned for results later on!

Works Cited:

Gruschczyk, B., Kamp, G., 1990. The shift from glycogenolysis to glycogen resynthesis after escape swimming: studies on the abdominal muscle of the shrimp, Crangon crangon. J Comp Physiol B, 753-760.

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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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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