BMA, our potential superheroes…pending

Connor Graham, Francis Marion University

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Findings: At the beginning of this summer my mentors and I had specific objectives and questions we wanted to answer regarding the biogeography of benthic microalgae and of course like any experimental hypotheses, things change. Our main objective was to identify the community structure on five barrier islands on South Carolina’s coast and see if there were differences. If there were differences were they because of geographic distance or environmental factors?  As the summer progressed our questions changed slightly to look more at community biomass instead. Of course our questions link back to the larger picture of using these diatoms as bioindicators for environmental health.

Community structure is composed of two main components: biomass and DNA composition. Biomass is the mass of the organisms present in a given area. Even though we collected samples for DNA, we had an allotted time which only allowed for analyzation of the biomass samples which were chlorophyll a. So, now our main questions were: Are there differences in community biomass among islands? Are those differences due to geographic distance or environmental factors like water temperature, nutrients, wind, pressure and so many more.

Based on the results from the data we have, biomass does indeed differ among islands, geographic distance is not the reason, but instead a few environmental factors. Those significant environmental factors are located in the table below. Still taking in account that we have pending analysis for DNA composition, nutrients and grain size, our original questions could be supported quite differently.

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The result of an ANOVA test which showed biomass differences among islands. The p-value was less than 0.0001.

 

 

 

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This p-value of 0.439 shows that Geographic distance is not correlated with community BMA biomass.

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These are the significant environmental factors that correlate with BMA biomass, with water temperature being the most significant with a p-value of 0.001.

However, if we do see that community structure is not affected by the differences in locations, then potentially there is no dispersal limitation on our microbes. Also, if community structure is also impacted by environmental like biomass, then we could potentially use this to measure bioindication by adding in a new factor.

As of now, we are not sure if diatoms can be used as bioindicators, and if they are the superheroes we need. However, we do know that more research is needed to find out and until then our great state awaits its savior.

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A picture of me covered in mud at Hunting Island after a day of sampling. Photo: Max Cook

Acknowledgements

I would like to thank my mentors: Dr. Craig Plante and Kristina Hill-Spanik (CofC). Also, I would like to thank my lab partner Max Cook (CofC). This project is supported by the Fort Johnson REU Program, NSF DBI-1757899.

 

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Not all superheroes wear capes!

Connor Graham, Francis Marion University

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The problem: When you think of superheroes, does the man in the red cape and ‘S’ on his chest come to mind? That’s understandable, but could it be possible that our greatest protectors are embedded in the sediment along our saltmarshes? Well, it is and these potential protectors are known as Benthic diatoms.

Benthic diatoms, plant-like microorganisms, are bioindicators, which means they can be used to determine the health of an environment. In South Carolina, environmental health is crucial to the prospering tourist areas, booming commercial fishing, and overall human health of the year-round residents. Poor environmental health could lead to a decline in economic benefits, decrease in seafood-and-shellfish heavy diets, and the fitness of the human population living in those areas. Benthic microalgae (BMA) are considered to be great bioindicators because of they have a short lifespan, they are abundant, easy to sample, sessile, and respond to specific stimuli (Desrosiers et al. 2013). But the question is can we use diatoms as bioindicators for South Carolina’s various salt marshes? Are they the superheroes we did not even know we had?

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Sampling site at Folly Beach. Photo: Max Cook.

My project this summer consists of sampling saltmarsh mud on at least five barrier islands along South Carolina’s coast to better understand the biogeography of BMA and assess their potential as bioindicators for saltmarshes. Barrier islands are land areas that are now inhabited by humans that protect inland territories from natural disasters.

I am comparing the community structure of the BMA’s on the various islands. If there is little to no variation in the benthic microbial communities gathered from the islands, bioindication can be used to determine their health. To use them as bioindicators will require the community structure to be similar on all the islands.

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Measuring the amount of light at Folly Beach. Photo: Max Cook.

Whether or not the community structure is similar or different will then be compared to the geographical distance of the sample sites and islands. Looking at the biogeography (geographical distribution of living things) of the BMA community has not been a priority, because we assume “everything is everywhere” (Baas-Becking 1934, as cited in Janne Soininen 2012) when speaking of microorganisms. Hopefully, by determining the diatoms’ community diversity on the islands, South Carolina is one step closer to thriving.

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Kristina, Max, and I in the clean room at Hollings Marine Lab analyzing grain sizes of sediment samples. Photo: Jennifer Ness.

Acknowledgments

I would like to thank my mentors: Dr. Craig Plante and Kristina Hill-Spanik (CofC). Also, I would like to thank my lab partner Max Cook (CofC). This project is supported by the Fort Johnson REU Program, NSF DBI-1757899.

Literature Cited:

Desrosiers, C., J. Leflaive., A. Eulin. and L. Ten-Hage. (2013) Bioindicators in marine waters: Benthic diatoms as a tool to assess water quality from eutrophic to oligotrophic coastal ecosystems. Ecological Indicators. 32: 25–34.

Soininen J. (2012) Macroecology of unicellular organisms – patterns and processes. Environmental Microbiology Reports, 4(1): 10-22.

The BMA of Today

Christine Hart, Clemson University

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In previous blog posts I described the sand-dwelling microalgae, also known as benthic microalgae (BMA), which are essential to estuary ecosystems. Not only do they produce the air we breathe and food we eat, they also inform us about the subtle changes that are occurring in our environment. Changes that otherwise may go unnoticed.

How do BMA show these environmental changes? By forming the foundation of estuarine energy, they provide a snapshot of how the estuary is functioning as a whole. If changes occur in BMA patterns, this may indicate changes in the overall ecosystem. BMA are also easily characterized and compared using modern molecular approaches. These qualities make BMA living indicators, or bioindicators, that are important in monitoring future ecosystem health.

BMA become visible in the upper layers of sediment at low tide. Later, they decrease in density—or biomass—as the tide rises. Our project studied the mechanism for the increase of biomass during low tide. Previous studies suggested that the mechanism for biomass increase is vertical migration of BMA from lower layers to upper layers of sediment. We also tested whether BMA growth due to high light exposure contributes to the biomass increase.

Our results indicated that both vertical migration and growth due to sunlight exposure were important to the increase in biomass. This is the first contribution to literature that recognizes a multifaceted approach to BMA biomass changes.

Additionally, we studied in how the biomass increase was connected to patterns in the type of BMA in Charleston Harbor. Previous studies suggested that increasing biomass was connected to changes in the abundance of BMA species; therefore, we expected to see the amount of certain BMA species change based on their exposure to migration and sunlight.

We were surprised by our findings. In this study, we found that BMA did not vary over short time periods (by tidal stage or by exposure to migration and sunlight). Instead, we found that BMA varied spatially and over a period of 6 years. In fact, only one of the dominant species of BMA remained the same from 2011 to 2017 (Figure 1).  The long-term change in community coincides with geological changes in the sampling site (Figure 2).

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Figure 1. The relative abundance of each dominant BMA species from 2011 to 2017 is shown immediately after sediment exposure (T0) and 3 hours later (TF). Only one species—Halamphora coffeaeformis—remains dominant in 2017. This is evidence of a dramatic change in the dominant type of BMA in Grice Cove.

These are positive results for the use of BMA as bioindicators. If types of BMA are invariable over short periods of time, measurements of BMA will be more precise. Bioindicators must be capable of showing changes that are occurring on a larger environmental scale; therefore, it would be a good sign if the change in BMA community reflects the changing geological environment (Figure 2). Still, more studies on the temporal and spatial patterns of BMA communities should be conducted before BMA can be used as bioindicators.

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Figure 2. Aerial view of Grice Cove sampling site over time. The approximate location of the sampling site is shown by the white line. Sampling sandbar has changed over time, possibly contributing to community changes. Source: “Grice Cove” 32 degrees 44’58”N 79 degrees 53’45”W. Google Earth. January 2012 to March 2014. June 20, 2017.

This study contributed new information to the studies of BMA biomass during low tide, and showed that the BMA of today in Grice Cove are significantly different than in previous years.

 

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 College of Charleston’s Grice Marine Laboratory.

 

Literature Cited:

Holt, E. A. & Miller, S. W. (2010) Bioindicators: Using Organisms to Measure Environmental Impacts. Nature Education Knowledge 3(10):8.

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.

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.

Searching in the Sand

Christine Hart, Clemson University

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In “Exploring the Secret Garden” I discussed our studies of the benthic microalgae (BMA) that inhabit the intertidal regions of beaches. The goal of our study is to identify the mechanisms involved in the visually noticeable increase of BMA during low tide. This mechanism will be linked to changes in the type of BMA dominating the sand flat. To accomplish these goals our study will incorporate field work, molecular techniques, and DNA analysis.

During field work we will collect and manipulate sediment to distinguish between an increase in BMA by either vertical migration or growth mechanisms. The sediment will be collected on a sand flat in Grice Cove (Figure 1). Sand will be sampled using corers, which pick up a layer of sand without disturbing the vertical organization. The collected sand will be split between measurements of biomass, or BMA density, and DNA analysis. Biomass is measured by finding the concentration of chlorophyll a in the sediment. BMA synthesize chlorophyll a; therefore, the concentration of chlorophyll a is proportional to the density of BMA.

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Figure 1. Aerial view of Grice Cove sampling site with the approximate location of the 50 m sand flat transect site. Sampling sand flat is open to the Charleston Harbor. Source: “Grice Cove” 3244’58”N 7953’45”W. Google Earth. March 20, 2017. June 20, 2017.

The methods for field work are represented in Figure 2. There are two vertical migration treatments: filter and mesh. Filter treatments prevent vertical migration between cored and surrounding sediment. Mesh treatments permit vertical migration. If migration is important to the biomass increase, biomass measurements in mesh will be greater than in filter treatments. Filter and mesh treatments will also be exposed to shade and light conditions to interpret the impact of growth on biomass. Sunlight provides the energy necessary for BMA growth. Without sunlight growth will be limited. If growth is the mechanism of biomass increase, the shaded samples will have a lower biomass than the light exposed samples.

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Figure 2. Field work methods visualization. Locations of replicates along the 50 m transect are chosen using a random number generator and marked with flags. Random coordinates and a quadrat of 50 cm by 50 cm are used to determine where sediment will be sampled and treatments will be placed. Three controls (T0, TM, and TF) are taken at time intervals 1.5 hours apart after sand exposure. During TM and TF time points, samples are taken from the 4 treatments shown above: filter, mesh, filter + shade, and mesh + shade. Filter treatments prevent vertical migration, while mesh treatments permit vertical migration. Shaded and non-shaded filter and mesh treatments will be important in determining the role of sun exposure in biomass increase.

To link the mechanism of biomass increase to the BMA composition, we will use molecular techniques and analyze the DNA found in the sediment. DNA will be extracted from the sediment and amplified using a polymerase chain reaction (PCR). The DNA will be sequenced using High Throughput Ion Torrent technology. The results from sequencing will identify the BMA present at each time point and within each treatment. This information will link the mechanism of biomass increase to the changes in BMA composition. Our understanding of BMA dynamics will establish a basis for the BMA ecology in the Charleston Harbor. In the future, BMA dynamics could be compared to our study to assess changes caused by human influences in Charleston estuaries.

 

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 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.

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.

 

What’s living in the sand?

Jessie Lowry, Coker College

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Visible microalgae seen on the surface of wet sand at Folly Beach.

Next time you go to the beach this summer, I want you to think about the sand that you are walking on. Did you know that there are tons of microscopic photosynthetic organisms, aka microalgae, that live on the surface of sand? Before this summer, I didn’t know about these organisms either. Here is a picture of visible microalgae on the surface of the sand. Look for this next time you’re at the beach!

Microalgae communities in sand are made up of single-celled eukaryotic algae and cyanobacteria living in the top several millimeters of the sand (Miller et al., 1996). These organisms play important roles in ecosystem productivity and food chain dynamics, as well as in sediment properties, such as erodibility (Miller et al., 1996).

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Dr. Craig Plante and Jessie Lowry collect samples of sediment from Folly Beach. Photo credit: Kristy Hill-Spanik.

I am studying these microalgal communities and what factors influence community structure. For example, does pH, salinity, nutrients, or grain size shape microalgal community structure? Or does geographic distance shape communities? To answer these questions, I am collecting samples from Kiawah Island, Folly Beach, Isle of Palms, and Pawley’s Island, SC. We are measuring environmental variables at each location, and using molecular tools to study microalgal community structure.

I am extracting the DNA from samples collected, amplifying specific regions from these samples using polymerase chain reaction (PCR), and then we will be getting these regions sequenced using Ion Torrent technology. We will then use QIIME to determine how similar these benthic microalgal communities are.

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Jessie Lowry preparing samples for PCR, or polymerase chain reaction, which is used to make millions of copies of a piece of DNA.

Diatoms, a group of microalgae, have been proposed as bioindicators of environmental health (Desrosiers et al., 2013). Bioindicators are really cool because instead of telling a snapshot of an environmental condition, such as pH, temperature, or amount of oxygen in an environment, biological indicators reflect those changes and can give an idea of how the ecosystem is being affected. This research will further our knowledge of what factors shape benthic microalgal communities, and give a better understanding of these organisms as a potential bioindicator. In addition, this research will add to knowledge about the distribution of microorganisms, which is also not fully understood.

Learn more:

http://web.vims.edu/bio/shallowwater/benthic_community/benthic_microalgae.html

http://www.aims.gov.au/docs/research/water-quality/runoff/bioindicators.html

References

Desrosiers, C., Leflaive, J., Eulin, A., Ten-Hage, L. (2013). Bioindicators in marine waters: benthic diatoms as a tool to assess water quality from eutrophic to oligotrophic coastal ecosystems. Ecological Indicators, 32, 25-34.

Miller, D.C., Geider, R.J., MacIntyre, H.L. Microphytobethos: The ecological role of the “Secret Garden” of unvegetated, shallow-water marine habitats. Estuaries, 19(2A): 186-212.

Acknowledgements

Thank you so much to my mentors Dr. Craig Plante, and Kristy Hill-Spanik. This research is funded through the National Science Foundation and College of Charleston’s Grice Marine Lab.

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Benthic Microalgae Research – A day at the beach

Jessica Lowry, Coker College

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I’m Jessie Lowry, a rising senior Biology major at Coker College, which is located in the tiny, homy town of Hartsville, SC. I am really excited to be here in my hometown this summer with opportunity to do research through the College of Charleston REU at Ft. Johnson. My research project this summer that I am working with Dr. Craig Plante on is investigating what factors influence the communities of benthic microalgae, or photosynthetic microorganisms in sediment.

Before we begin researching what species of diatoms make up the benthic microalgal communities, we need to do some preliminary sampling to measure things like pH, salinity, temperature, grain size, and moisture, at the three local beaches where I will be sampling at.

I’m not sure what I had envisioned for doing research this summer, but what I did not expect was for it to be like a day at the beach! Yesterday, Dr. Craig Plante and I went to Isle of Palms and Folly Beach, and today we are going to Kiawah.

My mentor, Dr. Craig Plante carrying research supplies out to the water at Folly Beach, SC.

We took several samples of seawater and sediment at each beach and we will do tests back at the lab. Unfortunately, I will not be spending every day researching at the beach. It was really great to get some sand, salt, and sun during a day of research.

Samples of seawater and sediment from Isle of Palms and Folly Beach that we collected to measure pH, salinity, grain size, and moisture.

Samples of seawater and sediment from Isle of Palms and Folly Beach that we collected to measure pH, salinity, grain size, and moisture.

Also, the temperature was in the 90s, the water was slightly warmer than the air, and the sand at one point at Folly Beach was a scorching 120° F! Dipping our feet in the water definitely felt great.

Funding

This research is funded by the National Science Foundation Reseach Experience for Undergraduates program at College of Charleston’s Grice Marine Lab.

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