North Carolina high school students examine the phytoplankton microbiome at the BPC

June 17th, 2016 @   - 

The Outer Banks is a narrow strip of land jutting out into the Atlantic off the coast of North Carolina. Life on the sandbar, as many locals call it, is full of adventure and challenges. Loved by many and with a rich history, this area is known as the first English settlement, Graveyard of the Atlantic, and launching pad of the Wright Brothers. Inhabited by Blackbeard, wild horses, surfers, artists, anglers, and dreamers and visited each summer by thousands of tourists and the occasional nor’easter and hurricane, this rugged coastline is also home to the Phytofinders, a small group of students at First Flight High School (FFHS), Kill Devil Hills, NC, who share a passion for learning about and protecting the waters which surround their community.

The Phytofinders was founded in 2005 by a group of eager, ready-to-learn students with the help of their science teacher, Katie Neller, a marine science enthusiast. What began as a few kids interested in the ocean soon evolved into something special when the Oceanic Engineering Society of the Institute of Electrical and Electronics Engineers (IEEE/OES) got involved, initially providing financial support and, more recently, significant mentoring.

Now, each week, a large crew of Phytofinders ventures out in all kinds of weather and all seasons to Jennette’s Pier in Nags Head and The Army Corps of Engineers Field Research Facility (FRF) Pier in Duck, NC. There they collect samples of phytoplankton from the ocean using a tow net and water sampler. Meteorological and oceanographic measurements are documented and the samples brought back to FFHS. The next morning, before most students have even left for school, the samples are analyzed by microscopy and the presence of various target species is quantified and recorded.

Since 2010, the IEEE/OES has provided the Phytofinders with a $5,000 annual grant, which has helped significantly to advance the program, enriching capabilities and enabling more students to participate. The funds have been used to purchase equipment and to enable student travel to the IEEE/OES sponsored OCEANS conferences. The first group of FFHS students attended the Virginia Beach, VA, OCEANS Conference in 2012. More recently, a group of Phytofinders attended the 2015 OCEANS Conference in Washington, DC. In each case, students submitted abstracts for review by the technical committee, had papers accepted for publication in the Proceedings of the conference, and presented their research in the technical sessions.

“We all worked really hard on our research papers, and being able to present them at a real conference was really cool. We were the youngest to present, but our research was just as relevant,” senior Sam Weybright said.

Back at home, one species on which the Phytofinders keep a particularly watchful eye is Pseudo-nitzschia, a diatom that produces domoic acid, a neurotoxin. When consumed in large amounts, domoic acid can cause disorientation, hallucinations, seizures, and even lead to death. Because domoic acid is retained in food webs, this can have disastrous effects on many species: shrimp and zooplankton ingest the phytoplankton, whales eat the zooplankton, fish eat the shrimp, and birds, sharks, and humans eat the fish. When high levels of domoic acid are present, birds can forget where to fly, whales may beach themselves, and humans can get amnesiac shellfish poisoning.

The Phytofinders have twice been the first to detect a bloom of Pseudo-nitzschia, providing a warning before the toxin reached harmful levels. Most recently, in 2014, an elevated presence of Pseudo-nitzschia appeared in some of the students’ samples. After the team finished their analysis, water was bottled up and sent to National Oceanic and Atmospheric Administration (NOAA), who came to the same conclusion the students had: There was an imminent toxic bloom threatening the coast of the Outer Banks. Because NOAA had been made aware of the situation, they monitored the coastal ocean closely for any unusual activity, ready to provide a warning to the public if needed.

Dylan Owens and Meghan Savona of the First Flight High School's Phytofinders sampling off the Army Corps of Engineers Field Research Facility (FRF) Pier in Duck, NC. The students extracted genomic DNA for microbiome sequencing at the MBL. Three recent FFHS graduates, long-term members of the Phytofinders, visited the BPC in June to see how their samples are amplified and sequenced

Dylan Owens and Meghan Savona of the First Flight High School’s Phytofinders sampling off the Army Corps of Engineers Field Research Facility (FRF) Pier in Duck, NC. The students extracted genomic DNA for microbiome sequencing at the MBL. Three recent FFHS graduates, long-term members of the Phytofinders, visited the BPC in June to see how their samples are amplified and sequenced. Photo credit: Todd Morrison

Students are learning that partnerships with organizations like NOAA and OES are vital to the success of their research program. In April, with the support of IEEE/OES, Drs. Todd and Hilary Morrison, an oceanographer and a molecular biologist from Woods Hole, Massachusetts, paid a visit to the Outer Banks and spent two full days with the Phytofinders , working with the students to improve the program as a whole and to develop future initiatives. Todd is a Senior Ocean Engineer with the Woods Hole Group and Hilary is a Senior Scientist at the Marine Biological Laboratory.
The students took a day off from regular classes to collect phytoplankton samples from FRF and Jennette’s Pier with the Morrisons. The weather was less than ideal, with cold air and even colder wind, but everyone showed great perseverance and ingenuity and focused on successful sampling. The day began with a bus ride to Duck, NC, where the students collected phytoplankton using a tow net and water sampler. Usually, just three samples are taken 1,000 feet along the pier: surface, mid-level, and deep. On this occasion, the students attempted to take 9 samples: surface, mid-level, and deep samples from three different positions along the pier to see if there was any variation between the locations. High winds and waves prevented depth-controlled sampling. However the students were able to acquire near-surface samples from three different locations along the pier. The students also collected metadata, including conductivity, temperature, and depth (CTD) and Secchi disk casts (a measurement of water transparency). After the samples were taken, the group had an opportunity to learn a little more about FRF Pier, which is closed to the general public. Some scientists who work at the pier showed the students how they collect data, monitor the waves and tides, and why their research is important.
“I didn’t really realize how many different jobs there are that revolve around ocean research,” junior Lake Lige said. “It’s made me seriously consider going into a marine science field when I go to college.”

After they sampled from FRF, the group traveled to Jennette’s Pier in Nags Head, NC, to collect phytoplankton, following a similar sampling protocol (and fighting the wind throughout). All samples were carefully labeled and brought back to FFHS. There, one or more students examined each sample under a microscope and the observations were recorded. This sample processing has been relatively standard at FFHS for several years, but now, thanks to IEEE/OES support, something new is being added to the scientific repertoire of the Phytofinders, taking their sample analysis capabilities to a whole new level.

First Flight High School graduates Jodie Awtrey, Nathan Butcher, and Parks Kelly at the Keck Sequencing Facility.

First Flight High School graduates Jodie Awtrey, Nathan Butcher, and Parks Kelly at the Keck Sequencing Facility. Photo credit: Todd Morrison

Working under the guidance of Dr. Hilary Morrison, the FFHS students learned how to extract DNA from the phytoplankton in their samples. The first DNA samples were hand carried back to MBL and successfully PCR-amplified and sequenced (microbial community structure determined) with IEEE/OES financial support. Since the Morrisons’ visit, the Phytofinders have continued to extract DNA from their field samples. In mid-June, three students will travel to Woods Hole, where they will be hosted by the Morrisons and will participate in sequencing their newest samples. The Woods Hole trip is expected to become a yearly event, so that students will continue to be involved in this aspect of the ongoing research program.

Determination of the microbial diversity of the samples, particularly how the community structure may change with sampling location, over time, and with environmental conditions, promises to be a new tool for monitoring the environmental health of the waters of the Outer Banks. Hilary predicts that the sequencing results may eventually lead to a paper in a peer reviewed journal with some of the Phytofinders as authors. A paper is also planned for OCEANS 2018 in Charleston, NC.

The Phytofinders are looking optimistically into the future with some ambitious new endeavors now being developed. During the visit, the group also discussed ideas for improving the program with Dr. Todd Morrison. These include the development of an interactive, web-based SQL database, where all of their research results can be stored, organized, and readily queried. The goal is to improve phytoplankton collection and analysis methods, while teaching the students how to design, structure, and build the system. This work is expected to result in a paper at OCEANS Charleston. The students will also design and fabricate a new net tow system that will give them greater control of depth and position and some quantitative measure of fluid flux through the mouth of the phytoplankton net during sampling from the piers. This is also expected to lead to an OCEANS Charleston paper.

“Getting a solid plan for the next couple of years, for the database, new nets, and DNA collection, was really important. I think that it’ll really help further the program in the future,” junior Joe Sawin said.

At this writing, Ms. Neller and Drs. Todd and Hilary Morrison are helping to the students plan the first trips to Woods Hole. There, in June and again in August, two teams of Phytofinders will get hands-on experience helping to sequence the DNA that they previously extracted. The FFHS Phytofinders are enthusiastically embracing these opportunities and look forward to presenting their research results at OCEANS 2018 in Charleston, South Carolina, just two short years away.

–article by Meghan Savona

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