Haloarchaea: Life at the Brink

Ben Farmer, University of Kentucky

IMG_20180705_150556The problem: What comes to mind when you think of extreme environments? The freezing tundra of Antarctica, or maybe the fiery lava flows of a Hawaiian volcanic zone? Those particularly interested in marine science may think of the deep ocean, perhaps the Marianas Trench. Whichever drastic environment you think of, one fascinating thing ties all of these extremes together: life finds a way to thrive in each of them.

Earth is home to as many as 1 trillion species, and the bulk of them are microbes (Locey and Lennon 2016). Microbes that are adapted to live in conditions that are inhospitable to most life on Earth are called extremophiles. Archaea and bacteria, the two domains of life aside from eukaryotes, represent the majority of extremophiles. While archaea were long thought to be a type of bacteria since the two appear very similar, archaea are more closely related to humans. Archaea are an important model organism because they have forged a niche in just about every habitat imaginable. Hot springs in Yellowstone National Park were among the first locations where archaea were discovered and owe their vibrant colors to these microbes (Oren and Rodriguez-Valera 2001).

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Man-made salt pans of Bonaire, Dutch Caribbean, tinged pink by archaea. Halophiles dominate these artificial habitats. Credit: Benjamin van de Water, Flickr, 2009.

Haloarchaea are what I am studying this summer at the College of Charleston. Halo– is a prefix meaning “salt,” and haloarchaea are halophilic, or salt-loving. Perhaps the most famous location that haloarchaea have been found is in the Dead Sea – evidently not so dead after all. Haloarchaea are commonly found in water 10 times as salty as the ocean, in conditions known as hypersaline. Our goal is to investigate what adaptations have made that possible.

We know that the amino acid composition of halophiles is unusually acidic (Martin et al. 1999). Proteins of halophiles are therefore also unusually acidic, which allows their proteins to properly fold in hypersaline conditions. What we do not know is whether the expression of proteins can change at different salinities. Better understanding how proteins are adapted in haloarchaea lends itself to understanding extremophiles on a broader scale.

Mechanisms that allowed microbes to function in seemingly inhospitable environments were likely responsible for evolution of life on Earth (Rampelotto 2010). There are many habitats today that mimic extreme environments from both ancient history and current conditions on other planets, such as Mars. Martian soil is incredibly salty, a result of surface water that evaporated long ago (https://dornsife.usc.edu/labs/laketyrrell/life-in-hypersaline-environments/). Halophiles may have once lived in those hypersaline Martian waters. Therefore, knowledge that we gain about haloarchaea adaptations is valuable to our understanding of life both on Earth and elsewhere.


Acknowledgements

Many thanks to my mentor, Dr. Matthew Rhodes, who has introduced me to everything from cell culturing to python. This project is funded through the National Science Foundation and supported by the Fort Johnson REU Program, NSF DBI- 1757899.


 



References

Locey KJ, Lennon JT (2016) Scaling laws predict global microbial diversity. Proc Natl Acad Sci 113:5970–5975

Martin DD, Ciulla R a, Roberts MF (1999) Osmoadaptation in Archaea. Appied Enviromental Microbiol 65:1815–1825

Oren A, Rodriguez-Valera F (2001) The contribution of halophilic Bacteria to the red coloration of saltern crystallizer ponds. FEMS Microbiol Ecol 36:123–130

Rampelotto PH (2010) Resistance of microorganisms to extreme environmental conditions and its contribution to astrobiology. Sustainability 2:1602–1623

 

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Bacteria in the Ocean? That Eat Iron??

Lauren Rodgers, Rutgers University

Version 2The problem: Have you ever asked yourself, what is iron? It is an element? A rock? Some weird orange-ish substance? Is it the tool that you use to get the wrinkles out of clothes? And what does iron even do? Does it just sit there? Does anything eat it? Can we make things out of it? Iron is one of the most abundant elements on earth, yet not many people know much about the important role it plays in our lives.

Iron is more than just an element, or something found within a rock. It’s a nutrient, something necessary for the growth and metabolism of almost every living organism on Earth (Hedrich & Johnson, 2011). In the ocean, iron is found in two different forms, ferrous iron or Fe(II), which is soluble in water, and ferric iron or Fe(III), which is insoluble in water (Hedrich & Johnson, 2011). Because ferrous iron is soluble it is the form of iron that can be used by most organisms in the water (Hedrich & Johnson, 2011). This ferrous iron, however, is limited in the ocean despite its abundance in the Earth’s crust. In fact, Fe(II) is present only in incredibly small concentrations, making it a major limiting factor of growth for all of the plants and algae in the ocean. This is important because these plants and algae serve as the base of many food chains, so if there is a limitation on the growth of these organisms, it affects every other organism throughout the food chain. Though iron is an extremely important nutrient for many living organisms, it is still not well understood. One of the least understood aspects is how iron specifically cycles through different marine environments. Does it ever change form? Does anything add iron to the ocean? Does anything take iron out of the ocean? These questions bring us to Zetaproteobacteria.

Zetaproteobacteria is a recently discovered class of iron-oxidizing microbes. This just means that the bacteria eat iron in the form of Fe(II) and produce Fe(III) as a waste product (Emerson et al., 2007; Chiu et al., 2017). In fact, these waste products can take on the form of hollow tubes, also called tubular sheaths, or twisted stalks that you can see under the microscope!

 

Zetaproteobacteria were initially described in 2007 near hydrothermal vents, utilizing the large concentrations of Fe(II) that were present in the fluid that spewed from the vents (Emerson et al., 2007).

Iron Mat

Iron mat composed of Zetaproteobacteria on a lava rock near the submarine Loihi volcano. (A. Malahoff, Hawaii, Loihi Volcano, July 1988)

How do Zetaproteobacteria relate to the cycling of iron? 

Zetaproteobacteria, with their role in eating iron and transforming it from its soluble Fe(II) state into its insoluble Fe(III) form may have an important role in the cycling of iron through the environment, functioning as an important source of iron removal.

Since their discovery, Zetaproteobacteria have also been observed in many other habitats, including coastal estuarine habitats with lower levels of iron, similar to that of Charleston, SC. (Laufer et al., 2017; Chiu et al., 2017). Our study will try to identify if these Zetaproteobacteria are present in the muddy soils around Charleston, as well as measure the levels of Fe(II) and Fe(III) in the rivers where these bacteria may be found.

 

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Hopefully, through the study of the distribution of Zetaproteobacteria across the globe, including the chemical characteristics of the different environments that they inhabit, we may get a clearer picture of how iron cycles in aquatic environments and the role that these Zetaproteobacteria play.


I would like to thank my mentor, Dr. Heather Fullerton, for guiding me through this research. I would also like to thank the National Science Foundation for funding this research as well as the College of Charleston and Grice Marine Lab for their support.


References 

Chiu, B. K., Kato, S., McAllister, S. M., Field, E. K., & Chan, C. S. (2017). Novel pelagic iron-oxidizing Zetaproteobacteria from the Chesapeake Bay oxic-anoxic transition zone. Frontiers in Microbiology, 8(JUL), 1–16. https://doi.org/10.3389/fmicb.2017.01280

Emerson, D., Rentz, J. A., Lilburn, T. G., Davis, R. E., Aldrich, H., Chan, C. S., & Moyer, C. L. (2007). A novel lineage of proteobacteria involved in formation of marine Fe-oxidizing microbial mat communities. PLoS ONE, 2(8), e667. https://doi.org/10.1371/journal.pone.0000667

Hedrich, S., Schlömann, M., & Johnson, D. B. (2011). The iron-oxidizing proteobacteria. Microbiology,157(6), 1551–1564.

Laufer, K., Nordhoff, M., Halama, M., Martinez, R. ., Obst, M., Nowak, M., … Kappler, A. (2017). Microaerophilic Fe(II)-oxidizing Zetaproteobacteriaisolated from low-Fe marine coastal sediments – physiology and characterization of their twisted stalks. Applied and Environmental Microbiology, 83(February), AEM.03118-16. https://doi.org/10.1128/AEM.03118-16

Mori, J. F., Scott, J. J., Hager, K. W., Moyer, C. L., Küsel, K., & Emerson, D. (2017). Physiological and ecological implications of an iron- or hydrogen-oxidizing member of the Zetaproteobacteria, Ghiorsea bivora, gen. nov., sp. Nov. ISME Journal, 11(11), 2624–2636. https://doi.org/10.1038/ismej.2017.132

The BMA of Today

Christine Hart, Clemson University

2017-06-22 10.29.36

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).

QualitativeLvM-MS

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.

Changes in Grice Cove

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

Interim report picture

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.

Sampling Site.png

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.

Field Work Diagram.png

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

Interim report picture

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

DSC01976

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).

IMG_8018

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.

IMG_7918

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.

DSC01972Unknown-4Unknown-3

Making Renewable Energy An Even Cheaper Alternative!

Yoel Cortes-Pena, Georgia Institute of Technology

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Fig 1. Picture of me

I’m Yoel Cortes-Pena, a chemical engineering senior student at Georgia Tech and future scientist and entrepreneur.  My research interests lie in renewable energy and environmental sustainability. Additionally, although I am an engineering student during the day, I am also part of Hip-Hop culture at night. My hobbies include dancing, beatboxing and rapping. Here is a link to my channel. 

Through this blog, I want to share with you my research experience as part of the Fort Johnson Undergraduate Summer Research Program. When I received the acceptance letter, I was surprised and happy that I would be working with Dr. Harold May in Microbial Electrosynthesis. This new technology uses microbes to fix carbon dioxide and electrons from an electrode to produce fuels and highly valued chemicals such as hydrogen, methane and acetate.

IMG_20150621_214304886

Fig 2.Picture of Microbial Electrosynthesis Reactor. The graphite rod on the left is the cathode (electron donor) and the rod on the right is the anode (electron acceptor). The left side of the reactor is being sparged with CO2. The microbes, located on the left side of the reactor, are fixing the CO2 and producing hydrogen (visible bubbles) and acetate (dissolved in solution).

One of the many applications of microbial electrosynthesis includes the storage of energy without contributing to carbon emissions. Solar, wind and other renewable energy forms output a variable amount of energy that tends to exceed public demand, especially during off-peak hours. Consequently, this surplus electricity becomes stranded energy that cannot be used. Microbial Electrosynthesis can utilize this excess or stranded energy and store it in fuel, valorizing the use of renewable energy technology.

Acknowledgements

Dr. Harold May’s Enviromental Microbiology lab is affiliated to the Medical University of South Carolina (MUSC).This project is possible thanks to funding from the NSF College of Charleston Summer REU program and the Grice Marine Laboratory. Lab space and facilities are provided by the Hollings Marine Laboratory.

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