Source: OHIO STATE UNIVERSITY submitted to NRP
TOWARDS AN UNDERSTANDING OF FUNDAMENTAL CHEMICAL INTERACTIONS BETWEEN MICROORGANISMS OR VIRUSES AND INORGANIC SUBSTRATES IN THE ENVIRONMENT
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0217825
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Feb 1, 2009
Project End Date
Sep 30, 2014
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
OHIO STATE UNIVERSITY
1680 MADISON AVENUE
WOOSTER,OH 44691
Performing Department
Unknown Department
Non Technical Summary
1. By developing new imaging capabilities that combines the nanometer-scale resolving power of the scanning probe microscope with the chemical identification capabilities of a fluorescence microscope we will open up interesting possibilities for the use of this instrument in various fields of research, including material, geological, and life sciences. 2. Understanding how soil microorganisms utilize specific membrane-proteins for metal-oxide reduction and biomineralization will give us a greater insight into how these processes can affect the mobility of pollutants (e.g., uranium, chromium) in the subsurface environment. Furthermore, by understanding the molecular mechanism that microbes direct the synthesis of mineral nanoparticles we can develop an important paradigm for bioinspired materials synthesis that will provide enormous insight into the strategies of controlled crystal synthesis used by other organisms, including multi-cellular organisms, and provide the basis for bio-controlled approaches to synthesize tailor-made inorganic nanostructures for applications across a diverse span of technologies. 3. Determining the mechanism(s) by which microorganisms or viruses use proteins to recognize specific solid substrates to produce pathogenic biofilms on the solid support or bind to a mineral substrate in a manner that promotes the infectious nature of the microbe or virus. By understanding this process we will be able to help prevent environmental outbreaks with human or economic consequences.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
1024010110020%
1334010110020%
7234010110020%
7234030110110%
7237299110010%
1027299110110%
1027299110010%
Knowledge Area
133 - Pollution Prevention and Mitigation; 102 - Soil, Plant, Water, Nutrient Relationships; 723 - Hazards to Human Health and Safety;

Subject Of Investigation
4010 - Bacteria; 4030 - Viruses; 7299 - Research equipment and methods, general/other;

Field Of Science
1101 - Virology; 1100 - Bacteriology;
Goals / Objectives
The goal of this project is to understand how fundamental chemical interactions that occur within a biomolecule (e.g., protein), between biomolecules, or between a biomolecule and an inorganic surface drive the various molecular processes needed to sustain life and how these interactions in turn affect the environment and human health. To achieve this goal we have the following two objectives: 1. Develop novel methods of fluorescent and scanning probe microscopy to characterize organic-inorganic interactions on a single-cell and single-molecule level. 2. Isolate, identify, and characterize microbial (e.g., magnetotactic bacteria, Shewanella, Staphylococcus) and viral (e.g., prions from chronic wasting disease, scrapie) proteins that are involved in organic-inorganic processes such as dissimilatory metal reduction, biomineralization of nanominerals, development of environmental and/or pathogenic biofilms, and environmental transmission of microbial- or viral-based diseases.
Project Methods
Objective 1 - An integrated atomic force microscope (AFM; Veeco/Digital Instruments; Bioscope AFM and NanoScope IV controller) / confocal laser scanning microscope (Zeiss; Axiovert 200M and LSM 510 META) will be used to collect simultaneous fluorescence and force measurements between inorganic substrates and biomolecules or cells. This instrument is located in Dr. Lower's laboratory at The Ohio State University Columbus, Ohio. Microbial cells will be located on a cover slip by using transmitted light and a 100X/1.45 N.A. objective lens on the confocal laser scanning microscope and an AFM cantilever will be positioned directly over a monolayer patch of cells (or uniform layer of biomolecules) prior to collecting force data. Fluorescence images will be collected by exciting cells with a 458/488 nm Argon laser and collecting the emission on a photodiode detector after passing the emitted light through a filter. Briefly, the tip of a cantilever will be brought into contact with a cell or biomolecule (i.e., "approach" force curves) and then pulled from the substrate surface (i.e., "retraction" force curves). The raw-data will be collected as the output of the photodiode detector (which is directly proportional to the deflection of the cantilever) relative to the position of the tip, which is translated by a piezoelectric scanner. These raw data are plotted as so-called "voltage-displacement" curves that may then be converted into "force-distance" curves and analyzed with SPIP (Image Metrology) and Igor Pro (WaveMetrics) software. For direct imaging, scanning tunneling microscopy, tapping mode or phase contrast mode will be used to collect detailed image (i.e., nanometer-scale resolution) of the samples. In addition to imaging the protein-mineral sample using single-molecule recognition force microscopy, we will also use AFM and STM to collect force and tunneling measurements between single prion molecules and mineral/soil substrates. Objective 2 - Microbes will be grown under a variety of conditions (e.g., aerobic, anaerobic) and proteins will be purified from the cells for Objective 1 or for identification and characterization. By growing the microorganisms under different conditions and subsequently purifying membrane-associated proteins from these cells we will be able to identify proteins that are involved in specific biogeochemical processes (e.g., biomineralization) and understand the molecular mechanisms that these microbes are utilizing to live in a particular environment and, in turn, examine the environmental and societal consequences of their metabolic processes. Soluble cytoplasmic and peripheral proteins are first removed using the sodium carbonate solution and insoluble material collected by centrifugation. The resulting pellet, now enriched with membrane proteins, will be solubilized using a solution of much stronger solubilization reagents allowing for subsequent resolution of the membrane proteins using 2D gel electrophoresis. Protein spots were excised, in-gel tryptic digestion and mass spectrometry (MS) will be performed. Protein identification will be preformed using both automated and manual database searches.

Progress 02/01/09 to 09/30/14

Outputs
Target Audience: Undergraduate and graduate students and high-school and middle-school students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Below is a list of opportunities for training and professional development that this project allowed us to accomplish. 1. This grant allowed one female Latino student to earn a M.S. degree from The Ohio State University. 2. The grant allowed one female student to earn a B.S./M.S. degree from The Ohio State University. 3. This grant allowed one female student to earn a Ph.D. degree from The Ohio State University. 4. This grant allowed one male student to earn a Ph.D. degree from The Ohio State University. 5. This grant allowed two students to travel to Africia to isolate magnetotact bacteria. 6. This grant allowed 3 students to travel to 4 different international conferences to present their research. 7. This grant allowed 1 student to travel to UNLV to work with Prof. Dennis A. Bazylinski, who is perhaps the world's leading authority on magnetotactic bacteria. 8. This grant allowed 1 student to travel to Oregon and British Columbia to collect novel species of magnetotactic bacteria. 9. This grant allowed the PI to design, develop and offer a new course on Apple iTunes U, which is freely available to anyone in the world. 10. This grant has also help allow the PI to teach high-school students (approximately 15 per year) from Columbus, Ohio. 11. This grant has also help allow the PI to teach high-school students (approximately 15 per year) from Columbus, Ohio. The students earned undergraduate credit from The Ohio State University. How have the results been disseminated to communities of interest? We disseminated our results through peer-reviewed journal articles, review articles, presentations at international conferences, presentations at universities, and in the classes that the PI teaches at The Ohio State University. The PI has also taught students from Metro High School, Columbus, Ohio. The PI also develop and offers a new course on Apple iTunes U (Intro Environmental Science), which is freely available to anyone in the world. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Below is a list of major research findings that this project allowed us to accomplish. 1. We successfully cloned three Mms genes, mms6, mms7 and mms13 from Magnetospirillum magneticum using PCR, inserted them into an expression vector, expressed their protein products. 2. We successfully developed a protein purification protocol for recombinant proteins Mms6, Mms7 and Mms13. 3. We successfully synthesized antibodies for Mms6 and Mms 13. 4. We isolated the first magnetotactic bacteria from Pavilion Lake, British Columbia. 5. We isolated the first magnetotactic bacteria from Mickey Hot Springs, Oregon. 6. We successfully used atomic force microscopy to study Mms-protien-magnetite binding. 7. We successfully used atomic force microscopy to image single Mms proteins on magnetite. 8. We successfully used transmission electron microscopy and confocal fluorescence microscopy in combination with nano-gold conjugated Mms antibodies to determine that Mms6 and Mms13 are localized to the magnetosome membrane in magnetotactic bacteria. 9. We developed a novel, inexpensive and simple isolation and enrichment technique to isolate magnetotactic bacteria from the environment. 10. We published 10 journal articles, 3 book chapters. 11. We received funding for Two grants from the National Science Foundation for a total of ~800k. 12. We successfully graduated 2 M.S. students, 1 B.S. student and 1 Ph.D. student. 13. We taught 2 undergraduate courses and 1 graduate course each year.

Publications


    Progress 01/01/13 to 09/30/13

    Outputs
    Target Audience: University undergraduate and graduate students, university professors, scientists, high school students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? 1. Taught Introduction to Environmental Science (ENR2100) to 500 undergraduate students during the Sp13 semester at OSU. 2. Taught Environmental Molecular Sciences course to 7 graduate students at OSU. 3. Taught Introduction to Environmental Science (ENR2100) to 500 undergraduate students during the Au13 semester at OSU. 4. Chaired the 2013 Environmental Science Student Symposium on Nov. 12, 2013 with 625+ undergraduate and graduate student poster presentations at OSU. 5. Advised one Ph.D. student. 6. Advised one B.S./M.S. student. 7. Served at an adviser to 7 undergraduate students majoring in Environmental Science with a focus in Molecular Sciences. 8. Developed and taught an iTunes U course through Apple iTunes to 25,000+ students in 2013. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

    Impacts
    What was accomplished under these goals? 1. Isolated several new strains of magnetotactic bacteria from Pavilion Lake, British Columbia, Canada.This discovery of magnetotactic bacteria (MTB) in Pavilion Lake offers an opportunity to better understand the diversity of MTB habitats, the geobiological function of MTB in unique freshwater ecosystems, and search for magnetofossils contained within the lake’s microbialites. 2. Identified two genes/proteins responsible for mineralizing magnetite. The discovery that magnetite, which was precipitated in the presence of purified recombinant-Mms7 and Mms13 proteins, resembles magnetite that is synthesized in MTB in-vivo supports the hypothesis that Mms6, Mms7, Mms13 control biomineralization of nanomagneite in MTB. 3. Isolated several new strains of magnetotactic bacteria from Mickey Hot Springs in Oregon, USA. This was the first time that a MTB was isolated from a ecosystem that contained high concentrations of arsenic. Intellectional Merit: The molecular mechanism by which magnetotactic proteins (e.g., Mms) catalyze the formation of magnetite is largely unknown. If funded, this grant will allow us to identify key amino acids and catalytic sites in Mms proteins, and determine their role in controlling the size, shape and intrinsic magnetic properties of the resulting Fe3O4 crystals. This knowledge would allow for the exciting possibility that we could then fabricate nanometer scale, single domain magnetic crystals that are tuned to elicit a particular magnetic response. Broader impacts: Magnetic nanoparticles are increasing in demand in emerging areas of technology and medicine (e.g., high-density data storage, ferrofluidic devices, targeted drug delivery, tissue regeneration scaffolding). For most applications, ideal “nano-magnets” have uniform shapes and morphologies, narrow size distributions, high crystallinity, single domain magnetism, the ability to remain dispersed in physiological solutions, have a maximal magnetic moment per particle, and controlled magnetic anisotropy. Magnetotactic bacteria synthesize magnetic crystals that meet all of these challenges. We have, in essence, attempted to mimic billions of years of evolutionary perfection by developing a protocol(s) to synthesize the nanomagnets in-vitro.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2013 Citation: Brian H. Lower and Dennis A. Bazylinski, "The Bacterial Magnetosome, a Unique Prokaryotic Organelle". Biomineralization in magnetotactic bacteria, Journal of Molecular Microbiology and Biotechnology. Vol. 23, 63-80. 2013.
    • Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Zachery Oestreicher, Steven K. Lower, Dennis A. Bazylinski and Brian H. Lower. "Magnetotactic bacteria from Pavilion Lake, British Columbia." Frontiers in Microbiology. 2014. (In-press)
    • Type: Journal Articles Status: Under Review Year Published: 2014 Citation: Carmen Valverde-Tercedor, Lijun Chen, Zachery Oestreicher, F. Abad�a-Molina, M. Martinez-Bueno, Steven K. Lower, Brian H. Lower, Dennis A. Bazylinski, Concepcion Jimenez-Lopez. "Subcellular localization of the magnetosome protein MamC in the marine magnetotactic bacterium Magnetococcus marinus strain MC-1 using immunoelectron microscopy," submitted to Archives of Microbiology. 2014. (In-review)


    Progress 01/01/12 to 12/31/12

    Outputs
    OUTPUTS: Class taught Spring 2012. The Ohio State University, School of Environment and Natural (SENR) Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. Class taught Autumn 2012. The Ohio State University, SENR. Research in Natural Resources, ENR 8000, 2 credit hours. Class taught Autumn 2012. The Ohio State University, School of Environment and Natural (SENR) Resources. Introduction to Environmental Science, ENR 2100, 3 credit hours. Served as a graduate student advisor for one B.S./M.S. student, one M.S. student and one Ph.D. student at The Ohio State University. Serving as a graduate student committee member for five Ph.D. students at The Ohio State University. Dissemination: Class taught to 300 students. Spring 2012. The Ohio State University, School of Environment and Natural Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. Class taught to 300 students. Autumn 2013. The Ohio State University, School of Environment and Natural Resources. Introduction to Environmental Science, ENR 2100, 3 credit hours. Class taught to 24 students. Autumn 2013. The Ohio State University, School of Environment and Natural Resources. Research in Natural Resources, ENR 8000, 2 credit hours. Change in knowledge: We identified the subcellular location of two different proteins of magnetotactic bacteria. This finding has helped in our understanding of which proteins these bacteria use to synthesis nanometer-sized magnetic magnetite minerals. We demonstrated how to image these tiny magnets using atomic force microscopy. We developed a simple protocol to purify magnetotactic bacteria from freshwater sources. One M.S. student who was working on this project graduated from OSU in the summer of 2012. One Ph.D. student who was working on this project graduated from OSU in the winter of 2012. PARTICIPANTS: 1. Brian H. Lower, Assistant Professor, The Ohio State University, PI/Project Director. 2. Steven K. Lower, Associate Professor, The Ohio State University, PI. 3. Lijun Chen, PhD student, The Ohio State University, student researcher. 4. Lumarie Perez-Guzman, MS student, The Ohio State University, student researcher. 5. Zachery Oestreicher, PhD student, The Ohio State University, student researcher. 6. Dennis Bazylinski, Professor/Director, UNLV, Co-PI. 7. Concepcion Jimenez-Lopez, Universidad de Granada, Spain, Co-PI. 8. Carmen Valverde Tercedor, Universidad de Granada, Spain, PhD student. TARGET AUDIENCES: University undergraduate and graduate students, university professors, scientists, high school students. Efforts: Taught an undergraduate course at The Ohio State University, School of Environment and Natural Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. Taught a graduate course at The Ohio State University, School of Environment and Natural Resources. Research in Natural Resources, ENR 8000, 3 credit hours. Instructor for graduate/faculty seminar series during Autumn Quarter 2010 at The Ohio State University, School of Environment and Natural Resources, Seminar Series, ENR 880, 1 credit hour. Served as a graduate student advisor for one B.S./M.S. student, one M.S. student and one Ph.D. student at The Ohio State University. Served as a graduate student committee member for five Ph.D. students at The Ohio State University. PROJECT MODIFICATIONS: Not relevant to this project.

    Impacts
    This research resulted in the development of a protein cloning and purification protocol that can be used by other laboratories to clone and purify bacterial proteins that catalyze the synthesis of nanometer-sized magnetic minerals. The research also resulted in the development of a new bacteria isolation protocol that can be used in any aquatic ecosystem to isolate and enrich the sample for magnetotactic bacteria. The protocol is simple (3 steps) and inexpensive (less than $25) and so it can be used by any school, college or university to teach students as a simple experiment that will teach students about environmental microbiology. It can also be used as an efficient method to isolate magnetotactic bacteria from remote field sites where access to reagents, electricity or lab ware is limited.

    Publications

    • Zachery Oestreicher, Carmen Valverde-Tercedor, Lijun Chen, Concepcion Jimenez-Lopez, Dennis A. Bazylinski, Nadia N. Casillas-Ituarte, Steven K. Lower and Brian H. Lower (2012). Magnetosomes and magnetite crystals produced by magnetotactic bacteria as resolved by atomic force microscopy and transmission electron microscopy. Micron. Vol. 43.: 1331-1335.
    • Nadia N. Casillas-Ituarte, Brian H. Lower, Supaporn Lamlertthon, Vance G. Fowler and Steven K. Lower (2012). Dissociation rate constants of human fibronectin binding to fibronectin-binding proteins on living Staphylococcus aureus isolated from clinical patients. Journal of Biological Chemistry. Vol. 287.: 6693-6701.
    • Zachery Oestreicher, Steven K. Lower, Wei Lin and Brian H. Lower (2012). Collection, isolation and enrichment of naturally occurring magnetotactic bacteria from the environment. Journal of Visualized Experiments. Vol. 69:e50123.
    • Brian H. Lower and Dennis A. Bazylinski (2012). The bacterial magnetosome: a unique prokaryotic organelle. Journal of Molecular Microbiology and Biotechnology. (in-press).


    Progress 01/01/11 to 12/31/11

    Outputs
    OUTPUTS: Activities: 1. Class taught Spring 2011. The Ohio State University, School of Environment and Natural (SENR) Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. 2. Class taught Spring 2011. The Ohio State University, School of Environment and Natural (SENR) Resources. Environmental Molecular Sciences, ENR 894, 3 credit hours. 3. Class taught Autumn 2011. The Ohio State University, SENR. Research in Natural Resources, ENR 800, 3 credit hours. 4. Instructor for graduate/faculty seminar series during Autumn Quarter 2011 at The Ohio State University, SENR, Seminar Series, ENR 880, 1 credit hour. 5. Served as a graduate student advisor for one M.S. student and one Ph.D. student at The Ohio State University. Serving as a graduate student committee member for five Ph.D. students at The Ohio State University. Events: 1. Presentation. Zachery Oestreicher, Steven K. Lower and Brian H. Lower (August 9, 2011). Magnetotactic Bacteria Containing Phosphorus-Rich Inclusion Bodies. Presented at Microscopy Society of America Microscopy and Microanalysis 2011 Annual Meeting. Nashville, Tennessee. 2. Presentation. Brian H. Lower (April 26, 2011 - Invited Presentation). Using Atomic Force Microscopy to Study Mineral-Microbe Interactions. Presented at 2011 Materials Research Society Spring Meting, to Materials Research Society. San Francisco, California. 3. Conference Symposium. Brian H. Lower served as a Co-Chair for a symposium. April 24 - 27, 2011. Materials Research Society Spring Meting in San Francisco, California. Symposium KK: Microbial Life on Surfaces - Biofilm-Material Interactions. 4. Presentation. Brian H. Lower (May 5, 2011 - Invited Presentaton). Prokaryotes with Backbones. Presented at Department of Evolution, Ecology and Organismal Biology, The Ohio State University, to Department of Evolution, Ecology and Organismal Biology. The Ohio State University. Columbus, Ohio. Products: 1. A new Environmental Molecular Science Option for the B.S. degree in Environmental Science at The Ohio State University. New collaboration: 1. Dr. Concepcion Jimenez-Lopez from Universidad de Granada, Spain to study biomineralization. Dr. Jimenez-Lopez sent one of her students to work with Dr. Lower at OSU. 2. Dr. Deric Learman from Central Michigan University. Dr. Learman visited Dr. Lower's research group and gave a presentation to the School of Environment and Natural Resources at The Ohio State University. Dissemination: 1. Class taught to 155 students. Spring 2011. The Ohio State University, School of Environment and Natural Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. 2. Class taught to 15 students. Spring 2011. The Ohio State University, School of Environment and Natural Resources. Environmental Molecular Sciences, ENR 894, 3 credit hours. PARTICIPANTS: 1. Brian H. Lower, Assistant Professor, The Ohio State University, PI/Project Director. 2. Steven K. Lower, Associate Professor, The Ohio State University, PI. 3. Lijun Chen, PhD student, The Ohio State University, student researcher. 4. Lumarie Perez-Guzman, MS student, The Ohio State University, student researcher. 5. Zachery Oestreicher, PhD student, The Ohio State University, student researcher. 6. Dennis Bazylinski, Professor/Director, UNLV, Co-PI. 7. Concepcion Jimenez-Lopez, Universidad de Granada, Spain, Co-PI. 8. Carmen Valverde Tercedor, Universidad de Granada, Spain, PhD student. TARGET AUDIENCES: University undergraduate and graduate students, university professors, scientists, high school students. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

    Impacts
    Change in knowledge: We successfully purfied three novel proteins from a magnetotactic bacteria. This will enable other scientists who are studying mineral biomineralization to use this technique to clone genes and purify proteins for their research. We also successfully isolated a magnetotactic bacteria from a geothermal spring in Oregon. This is the first time that such an organism was isolated from a geothermal spring. This discovery demonstrates that these microbes inhabit hyperthermic aquatic ecosystems and encourages the exploration for other species of magnetotactic bacteria in other "extreme" environments. In addition Taught an undergraduate course at The Ohio State University, School of Environment and Natural Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. Dr. Lower served as a graduate student advisor for one M.S. student and one Ph.D. student at The Ohio State University. Served as a graduate student committee member for five Ph.D. students at The Ohio State University. Dr. Lower taught a new course titled Environmental Molecular Sciences (ENR894) for undergraduate and graduate students at The Ohio State University and Universidade Federal de Pernambuco, Recife, Brazil. This class allowed OSU students to learn about science in South America and interact with students from Brazil.

    Publications

    • Zachery Oestreicher, Carmen Valverde Tercedor, Lijun Chen, Concepcion Jimenez-Lopez, Dennis A. Bazylinski, Steven K. Lower and Brian H. Lower (2011). Achieving nanoscale resolution when studying protein-catalyzed biomineralization. Micron (In review).
    • Nadia N. Casillas-Ituarte, Brian H. Lower, Supaporn Lamlertthon, Vance G. Fowler and Steven K. Lower. (2011 - In Press) Dissociation rate constants of human fibronectin binding to fibronectin-binding proteins on living Staphylococcus aureus isolated from clinical patients. Journal of Biological Chemistry.
    • Steven K. Lower, Nadia Casillas-Ituarte, Supaporn Lamlertthon, Roberto Lins, Ruchirej Yongsunthon, Eric Taylor, Alex DiBartola, Catherine Edmonson, Lauren McIntyre, Brian H. Lower, and Vance Fowler (2011). Polymorphisms in fibronectin binding protein A of Staphylococcus aureus are associated with infection of cardiovascular devices. Proceedings of the National Academy of Sciences USA 108:18372-18377.
    • Brian H. Lower and Steven K. Lower (2011). Force Spectroscopy of Mineral-Microbe Bonds. In Life at the Nanoscale: Atomic Force Microscopy of Live Cells, p. 301-316. Edited by Yves F. Dufrene. Singapore: Pan Stanford Publishing Pte Ltd.


    Progress 01/01/10 to 12/31/10

    Outputs
    OUTPUTS: Activities: Spring 2010. The Ohio State University, School of Environment and Natural (SENR) Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. Taught a graduate course at The Ohio State University, SENRs. Research in Natural Resources, ENR 800, 3 credit hours. Instructor for graduate/faculty seminar series during Autumn Quarter 2010 at The Ohio State University, SENR, Seminar Series, ENR 880, 1 credit hour. Serving as an graduate student advisor for one M.S. student and one Ph.D. student at The Ohio State University. Serving as a graduate student committee member for two Ph.D. students at The Ohio State University. Events: Presentations. Brian H. Lower (December 3, 2010 - Invited Presentation). "Biogeochemistry on a Molecular Scale." School of Life Sciences, University of Nevada, Las Vegas, Nevada. Lijun Chen and Brian H. Lower (September 1-4, 2010). "Using Single-Molecule Atomic Force Microscopy to Study Magnetite Biomineralization in Magnetotactic Bacteria. "The Second International Symposium on Magnetotactic Bacteria and Biomineralization, Beijing, China. Services: EMSL User Advisory Committee. Government. U.S. Department of Energy, Environmental Molecular Science Laboratory. Pacific Northwest National Laboratory. Richland, Washington, USA. EMSL Geochemistry, Biogeochemistry & Subsurface Science Science Review Panal. Government. U.S. Department of Energy, Environmental Molecular Science Laboratory. Pacific Northwest National Laboratory. Richland, Washington, USA. August 12-14, 2009. Products: A new Environmental Molecular Science Option for the B.S. degree in Environmental Science at The Ohio State University. New collaboration with Dr. Dennis Bazylinski from UNLV to study biomineralization. Dr. Brian Lower was awarded a three-year, $404,000 grant from the National Science Foundation to study the molecular mechanism of nanomagnetite synthesis in bacteria. Developed a new website to showcase our research and accomplishments: http://web.me.com/brianlower/Homepage/Welcome.html . Dissemination: Spring 2010. The Ohio State University, School of Environment and Natural Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. PARTICIPANTS: Brian H. Lower, Assistant Professor, The Ohio State University, PI/Project Director. Steven K. Lower, Associate Professor, The Ohio State University, PI. Lijun Chen, PhD student, The Ohio State University, student researcher. Lumarie Perez-Guzman, MS student, The Ohio State University, student researcher. Zachery Oestreicher, PhD student, The Ohio State University, student researcher. Dennis Bazylinski, Professor/Director, UNLV, Co-PI. TARGET AUDIENCES: Target Audiences: University undergraduate and graduate students, university professors, high school students. Efforts: Summer student internship for one high school student from Olentangy School District, Ohio during the summer of 2010. Taught an undergraduate course at The Ohio State University, School of Environment and Natural Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. Taught a graduate course at The Ohio State University, School of Environment and Natural Resources. Research in Natural Resources, ENR 800, 3 credit hours. Instructor for graduate/faculty seminar series during Autumn Quarter 2010 at The Ohio State University, School of Environment and Natural Resources, Seminar Series, ENR 880, 1 credit hour. Served as an graduate student advisor for one M.S. student and one Ph.D. student at The Ohio State University. Served as a graduate student committee member for two Ph.D. students at The Ohio State University. Developed a new course titled Environmental Molecular Sciences for undergraduate and graduate students at The Ohio State University. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

    Impacts
    Change in knowledge: Developed a new method for cloning genes from magnetotactic bacteria. We successfully cloned three genes (mms5, mms7, and mms13) from a magnetotactic bacteria. This will enable other scientists who are studying mineral biomineralization to use this technique to clone genes for their research. We also successfully isolated a magnetotactic bacteria from the Olentangy River in Columbus, Ohio. To my knowledge, this is the first time that such an organism was isolated from this river. This discovery demonstrates that these microbes inhabit freshwater ecosystems in Ohio and encourages the exploration for other species of magnetotactic bacteria in other Ohio watersheds.

    Publications

    • Lijun Chen, Dennis A. Bazylinski and Brian H. Lower. 2010. Bacteria that Synthesize Nano-sized Compasses to Navigate Earth. Nature Education Knowledge. Vol. 1, no. 10. : 14.
    • Lumarie Perez-Guzman, Kyle R. Bogner and Brian H. Lower. 2010. Earth's Ferrous Wheel. Nature Education Knowledge. Vol. 1, no. 10. : 8.
    • Steven K. Lower, Ruchirej Yongsunthon, Alex C. DiBartola, Brian H. Lower, Andrew W. Buck and Vance G. Fowler. 2010. A Tactile Response in Staphylococcus aureus. Biophysical Journal. Vol. 99, no. 9. : 2803-2811.
    • Brian H. Lower. 2011. Antibody recognition force microscopy (Ig-RFM) to identify and map the nanoscale distrubution of protein molecules on the surface of live microorganisms. In Microscopy: Science, Technology, Applications and Education. Edited by A. Mendez-Vilas and J. Diaz. Badajoz, Spain: Formatex Research Center.


    Progress 01/01/09 to 12/31/09

    Outputs
    OUTPUTS: Activities: Spring 2009. The Ohio State University, School of Environment and Natural (SENR) Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. Taught a graduate course at The Ohio State University, SENRs. Research in Natural Resources, ENR 800, 3 credit hours. Instructor for graduate/faculty seminar series during Autumn Quarter 2009 at The Ohio State University, SENR, Seminar Series, ENR 880, 1 credit hour. Serving as an graduate student advisor for one M.S. student and one Ph.D. student at The Ohio State University. Serving as a graduate student committee member for two Ph.D. students at The Ohio State University. Events: Poster Presentation. Zachery W. Oestreicher, Brian H. Lower, Roberto D. Lins, and Steven K. Lower. Peptides as Templates for Biomineralization. Presented at American Society for Microbiology 109th General Meeting. Phildelphia, Pennsylvania, USA. May 17 - 21, 2009. Services: EMSL User Advisory Committee. Government. U.S. Department of Energy, Environmental Molecular Science Laboratory. Pacific Northwest National Laboratory. Richland, Washington, USA. EMSL Geochemistry, Biogeochemistry & Subsurface Science Science Review Panal. Government. U.S. Department of Energy, Environmental Molecular Science Laboratory. Pacific Northwest National Laboratory. Richland, Washington, USA. August 12-14, 2009. Products: A new Environmental Molecular Science Option for the B.S. degree in Environmental Science at The Ohio State University. New collaboration with Dr. Colleen Hansel from Harvard University to study biomineralization. New collaboration with Dr. Dennis Bazylinski from UNLV to study biomineralization. Dr. Brian Lower was awarded a three-year, $404,000 grant from the National Science Foundation to study the molecular mechanism of nanomagnetite synthesis in bacteria. Developed a new website to showcase our research and accomplishments: http://web.me.com/brianlower/Homepage/Welcome.html . Dissemination: Spring 2009. The Ohio State University, School of Environment and Natural Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. Columbus Dispatch, April 5, 2009 Sunday Edition Science Article titled "Tricking Toxins". http://www.dispatch.com/live/content/science/stories/2009/04/05/sci_t oxbacteria.ART_ART_04-05-09_G3_QKDEBCE.htmlsid=101 Discovery Channel News, online news article, March 27, 2009 titled "Microbes' Toxic Metal-Digestion Secret Revealed". http://dsc.discovery.com/news/2009/03/27/bacteria-uranium.html MSNBC News Article, March 27, 2009 titled "Microbes digest, detoxify dangerous metals". http://www.msnbc.msn.com/id/29915669 The Ohio State University Research News Article posted online on their homepage http://researchnews.osu.edu/archive/bactmetal.htm Applied and Environmental Journal Cover, Vol. 75, Issue 9. Department of Energy News Article for May 2009 titled "Tips for Nanomapping". http://www.emsl.pnl.gov/news/highlights/lower20090601.pdf Pacific Northwest National Laboratory June 2009 News Article titled "Breathing, Bacterium's Role in Remediation". http://www.pnl.gov/science/highlights/highlight.aspid=616 PARTICIPANTS: Brian H. Lower, Assistant Professor, The Ohio State University, PI/Project Director. Steven K. Lower, Associate Professor, The Ohio State University, PI. Lijun Chen, PhD student, The Ohio State University, student researcher. Lumarie Perez-Guzman, MS student, The Ohio State University, student researcher. Zachery Oestreicher, PhD student, The Ohio State University, student researcher. Dennis Bazylinski, Professor/Director, UNLV, Co-PI. Roberto D. Lins, Associate Professor, Universidade Federal de Pernambuco, Recife, Brazil, Co-PI. Jean-Francois Boily, Associate Professor, Umea University, Sweden, Co-PI. Hermann Gruber, Professor, Johannes Kepler Universitat, Linz, Austria, Co-PI. Liang Shi, Senior Scientist, Pacific Northwest National Laboratory, Co-PI. Michael F. Hochella, Professor, Virginia Tech, Co-PI. Saumyaditya Bose, Post Doctoral Fellow, University of California Berkeley, Co-PI. Yuri A. Gorby, Senior Scientist, J. Craig Venter Institute, Co-PI. Andrew S. Madden, Assistant Professor, University of Oklahoma, Co-PI. TARGET AUDIENCES: Target Audiences: University undergraduate and graduate students, university professors, high school students. Efforts: Summer student internship for one high school student from Olentangy School District, Ohio during the summer of 2009. Taught an undergraduate course at The Ohio State University, School of Environment and Natural Resources. Introduction to Environmental Science, ENR 201, 5 credit hours. Taught a graduate course at The Ohio State University, School of Environment and Natural Resources. Research in Natural Resources, ENR 800, 3 credit hours. Instructor for graduate/faculty seminar series during Autumn Quarter 2009 at The Ohio State University, School of Environment and Natural Resources, Seminar Series, ENR 880, 1 credit hour. Served as an graduate student advisor for one M.S. student and one Ph.D. student at The Ohio State University. Served as a graduate student committee member for two Ph.D. students at The Ohio State University. Developed a new course titled Environmental Molecular Sciences for undergraduate and graduate students at The Ohio State University. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

    Impacts
    Change in knowledge: Developed a new method of imaging single protein molecules called antibody recognition force microscopy that was published in the journal Applied and Environmental Microbiology, 75: 2931-2935. This method will allow scientists in biology and geology to image live cells in-situ with nanometer resolution. Showed that a soil microorganism called Shewanella oneidensis synthesizes and targets specific proteins to the bacterium's outer surface when the proteins function to "detoxify" dangerous metals, such as uranium and technetium, that may be present in the subsurface environment. This information will lead to the development of new bioremediation strategies for cleaning up DOE superfund sites that are contaminated with radionuclides and heavy metals.

    Publications

    • Brian H. Lower, Ruchirej Yongsunthon, Liang Shi, Linda Wildling, Hermann J. Gruber, Nicholas S. Wigginton, Catherine L. Reardon, Grigoriy E. Pinchuk, Timothy C. Droubay, Jean-Francois Boily, and Steven K. Lower. 2009. Antibody-Recognition Force Microscopy shows that Outer Membrane Cytochromes OmcA and MtrC are Expressed on the Exterior Surface of Shewanella oneidensis MR-1. Applied and Environmental Microbiology. Vol. 75: 2931-2935.
    • Saumyaditya Bose, Michael F. Hochella Jr., Yuri A. Gorby, David W. Kennedy, Dave E. McCready, Andrew Madden, and Brian H. Lower. 2009. Bioreduction of Hematite Nanoparticles by the Dissimilatory Iron Reducing Bacterium Shewanella oneidensis MR-1. Geochimica et Cosmochimica Acta. Vol. 73: 962-976.