Separation of Bacteria, Protozoa and Carbon Nanotubes by Density Gradient Centrifugation

Mortimer, Monika; Petersen, Elijah; Buchholz, Bruce; Holden, Patricia

doi:10.3390/nano6100181

Title: Separation of Bacteria, Protozoa and Carbon Nanotubes by Density Gradient Centrifugation

Journal Article · Wed Oct 12 00:00:00 EDT 2016 · Nanomaterials

DOI:https://doi.org/10.3390/nano6100181· OSTI ID:1377787

Mortimer, Monika ^[1]; Petersen, Elijah ^[2]; Buchholz, Bruce ^[3]; Holden, Patricia ^[4]

Univ. of California, Santa Barbara, CA (United States). Bren School of Environmental Science and Management, Earth Research Inst., and Center for the Environmental Implications of Nanotechnology; National Institute of Chemical Physics and Biophysics, Tallinn (Estonia). Lab. of Environmental Toxicology
National Inst. of Standards and Technology (NIST), Gaithersburg, MD (United States). Biosystems and Biomaterials Division, Material Measurement Lab.
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Univ. of California, Santa Barbara, CA (United States). Bren School of Environmental Science and Management, Earth Research Inst., and Center for the Environmental Implications of Nanotechnology

Sustainable production and use of carbon nanotube (CNT)-enabled materials require efficient assessment of CNT environmental hazards, including the potential for CNT bioaccumulation and biomagnification in environmental receptors. Microbes, as abundant organisms responsible for nutrient cycling in soil and water, are important ecological receptors for studying the effects of CNTs. Quantification of CNT association with microbial cells requires efficient separation of CNT-associated cells from individually dispersed CNTs and CNT agglomerates. Here in this paper, we designed, optimized, and demonstrated procedures for separating bacteria (Pseudomonas aeruginosa) from unbound multiwall carbon nanotubes (MWCNTs) and MWCNT agglomerates using sucrose density gradient centrifugation. We demonstrate separation of protozoa (Tetrahymena thermophila) from MWCNTs, bacterial agglomerates, and protozoan fecal pellets by centrifugation in an iodixanol solution. The presence of MWCNTs in the density gradients after centrifugation was determined by quantification of ¹⁴C-labeled MWCNTs; the recovery of microbes from the density gradient media was confirmed by optical microscopy. Protozoan intracellular contents of MWCNTs and of bacteria were also unaffected by the designed separation process. Lastly, the optimized methods contribute to improved efficiency and accuracy in quantifying MWCNT association with bacteria and MWCNT accumulation in protozoan cells, thus supporting improved assessment of CNT bioaccumulation.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)

Sponsoring Organization:: USDOE: National Science Foundation (NSF); USEPA; Estonian Research Council

Grant/Contract Number:: AC52-07NA27344; PUTJD16

OSTI ID:: 1377787

Report Number(s):: LLNL-JRNL-684658; NANOKO; PII: nano6100181

Journal Information:: Nanomaterials, Vol. 6, Issue 10; ISSN 2079-4991

Publisher:: MDPICopyright Statement

Country of Publication:: United States

Language:: English

Citation Metrics:

Cited by: 15 works

Citation information provided by
Web of Science

References (44)

Ecological Nanotoxicology: Integrating Nanomaterial Hazard Considerations Across the Subcellular, Population, Community, and Ecosystems Levels Holden, Patricia A.; Nisbet, Roger M.; Lenihan, Hunter S. Accounts of Chemical Research, Vol. 46, Issue 3 https://doi.org/10.1021/ar300069t	journal	July 2012
Exposure to CuO Nanoparticles Changes the Fatty Acid Composition of Protozoa Tetrahymena thermophila Mortimer, Monika; Kasemets, Kaja; Vodovnik, Maša Environmental Science & Technology, Vol. 45, Issue 15 https://doi.org/10.1021/es201524q	journal	August 2011
A simple separation method for downstream biochemical analysis of aquatic microbes Garrison, Cody E.; Bochdansky, Alexander B. Journal of Microbiological Methods, Vol. 111 https://doi.org/10.1016/j.mimet.2015.01.025	journal	April 2015
“Density equilibrium” method for the quantitative and rapid in situ determination of lipid, hydrocarbon, or biopolymer content in microorganisms Eroglu, Ela; Melis, Anastasios Biotechnology and Bioengineering, Vol. 102, Issue 5 https://doi.org/10.1002/bit.22182	journal	April 2009
The effects of high energy probe sonication on the thermoelectric power of large diameter multiwalled carbon nanotubes synthesized by chemical vapor deposition Hewitt, Corey A.; Craps, Matt; Czerw, Richard Synthetic Metals, Vol. 184 https://doi.org/10.1016/j.synthmet.2013.09.026	journal	November 2013
Toxicity of 12 metal-based nanoparticles to algae, bacteria and protozoa Aruoja, Villem; Pokhrel, Suman; Sihtmäe, Mariliis Environmental Science: Nano, Vol. 2, Issue 6 https://doi.org/10.1039/C5EN00057B	journal	January 2015
Use of the Sucrose Gradient Method for Bacterial Cell Cycle Synchronization Choudhary, Abha; Lin, Lin; Choudhary, Madhusudan Journal of Microbiology & Biology Education, Vol. 13, Issue 1 https://doi.org/10.1128/jmbe.v13i1.346	journal	May 2012
Five reasons to use bacteria when assessing manufactured nanomaterial environmental hazards and fates Holden, Patricia A.; Schimel, Joshua P.; Godwin, Hilary A. Current Opinion in Biotechnology, Vol. 27 https://doi.org/10.1016/j.copbio.2013.11.008	journal	June 2014
Carbon nanotube clusters as universal bacterial adsorbents and magnetic separation agents Moon, Hyung-Mo; Kim, Jin-Woo Biotechnology Progress https://doi.org/10.1002/btpr.294	journal	January 2009
Quantification of Carbon Nanotubes in Environmental Matrices: Current Capabilities, Case Studies, and Future Prospects Petersen, Elijah J.; Flores-Cervantes, D. Xanat; Bucheli, Thomas D. Environmental Science & Technology, Vol. 50, Issue 9 https://doi.org/10.1021/acs.est.5b05647	journal	April 2016
Effects of TiO ₂ and Ag Nanoparticles on Polyhydroxybutyrate Biosynthesis By Activated Sludge Bacteria Priester, John H.; Van De Werfhorst, Laurie C.; Ge, Yuan Environmental Science & Technology, Vol. 48, Issue 24 https://doi.org/10.1021/es504117x	journal	November 2014
Bacterial remediation from effluent containing multi-walled carbon nanotubes Lemes, A. P.; Cordi, L.; Santos, A. Journal of Physics: Conference Series, Vol. 304 https://doi.org/10.1088/1742-6596/304/1/012023	journal	July 2011
Iodixanol: A Nonionic Iso-osmotic Centrifugation Medium for the Formation of Self-Generated Gradients Ford, T.; Graham, J.; Rickwood, D. Analytical Biochemistry, Vol. 220, Issue 2 https://doi.org/10.1006/abio.1994.1350	journal	August 1994
Determination of nanoparticle size distribution together with density or molecular weight by 2D analytical ultracentrifugation Carney, Randy P.; Kim, Jin Young; Qian, Huifeng Nature Communications, Vol. 2, Issue 1 https://doi.org/10.1038/ncomms1338	journal	June 2011
Bulk enrichment and separation of multi-walled carbon nanotubes by density gradient centrifugation Deng, Xiaoyong; Xiong, Dongmei; Wang, Haifang Carbon, Vol. 47, Issue 6 https://doi.org/10.1016/j.carbon.2009.02.016	journal	May 2009
Bioaccumulation of Multiwall Carbon Nanotubes in Tetrahymena thermophila by Direct Feeding or Trophic Transfer Mortimer, Monika; Petersen, Elijah J.; Buchholz, Bruce A. Environmental Science & Technology, Vol. 50, Issue 16 https://doi.org/10.1021/acs.est.6b01916	journal	July 2016
Carbon nanotube compared with carbon black: effects on bacterial survival against grazing by ciliates and antimicrobial treatments Chan, Tiffany S. Y.; Nasser, Fatima; St-Denis, Christine H. Nanotoxicology, Vol. 7, Issue 3 https://doi.org/10.3109/17435390.2011.652205	journal	January 2012
Bioaccumulation of Radio-Labeled Carbon Nanotubes by Eisenia foetida Petersen, Elijah J.; Huang, Qingguo; Weber, Jr., Walter J. Environmental Science & Technology, Vol. 42, Issue 8 https://doi.org/10.1021/es071366f	journal	April 2008
Relevance of octanol-water distribution measurements to the potential ecological uptake of multi-walled carbon nanotubes Petersen, Elijah J.; Huang, Qingguo; Weber, Walter J. Environmental Toxicology and Chemistry https://doi.org/10.1002/etc.149	journal	January 2010
High performance separation of aerosol sprayed mesoporous TiO2 sub-microspheres from aggregates via density gradient centrifugation Zhang, Yichi; Shi, Yifeng; Liou, Ya-Hsuan Journal of Materials Chemistry, Vol. 20, Issue 20 https://doi.org/10.1039/b926183d	journal	January 2010
Uptake, localization and clearance of quantum dots in ciliated protozoa Tetrahymena thermophila Mortimer, Monika; Kahru, Anne; Slaveykova, Vera I. Environmental Pollution, Vol. 190 https://doi.org/10.1016/j.envpol.2014.03.021	journal	July 2014
Degradation of multiwall carbon nanotubes by bacteria Zhang, Liwen; Petersen, Elijah J.; Habteselassie, Mussie Y. Environmental Pollution, Vol. 181 https://doi.org/10.1016/j.envpol.2013.05.058	journal	October 2013
Ultrasonic dispersion of nanoparticles for environmental, health and safety assessment – issues and recommendations Taurozzi, Julian S.; Hackley, Vincent A.; Wiesner, Mark R. Nanotoxicology, Vol. 5, Issue 4 https://doi.org/10.3109/17435390.2010.528846	journal	September 2010
Sucrose density gradient centrifugation separation of gold and silver nanoparticles synthesized using Magnolia kobus plant leaf extracts Lee, So Hyun; Salunke, Bipinchandra K.; Kim, Beom Soo Biotechnology and Bioprocess Engineering, Vol. 19, Issue 1 https://doi.org/10.1007/s12257-013-0561-4	journal	February 2014
Plasmolysis and Cell Shape Depend on Solute Outer-Membrane Permeability during Hyperosmotic Shock in E. coli Pilizota, Teuta; Shaevitz, Joshua W. Biophysical Journal, Vol. 104, Issue 12 https://doi.org/10.1016/j.bpj.2013.05.011	journal	June 2013
Effects of Soluble Cadmium Salts Versus CdSe Quantum Dots on the Growth of Planktonic Pseudomonas aeruginosa Priester, John H.; Stoimenov, Peter K.; Mielke, Randall E. Environmental Science & Technology, Vol. 43, Issue 7 https://doi.org/10.1021/es802806n	journal	April 2009
Colloidal stability and ecotoxicity of multiwalled carbon nanotubes: Influence of select organic matters: Nanomaterials in the environment Cerrillo, Cristina; Barandika, Gotzone; Igartua, Amaya Environmental Toxicology and Chemistry, Vol. 35, Issue 1 https://doi.org/10.1002/etc.3172	journal	November 2015
Dispersion of TiO2 Nanoparticle Agglomerates by Pseudomonas aeruginosa Horst, A. M.; Neal, A. C.; Mielke, R. E. Applied and Environmental Microbiology, Vol. 76, Issue 21 https://doi.org/10.1128/AEM.00324-10	journal	September 2010
Analytical Protocols for Separation and Electron Microscopy of Nanoparticles Interacting with Bacterial Cells Sousa, Cláudia; Sequeira, Diana; Kolen’ko, Yury V. Analytical Chemistry, Vol. 87, Issue 9 https://doi.org/10.1021/ac503835a	journal	April 2015
Use of Tetrahymena thermophila To Study the Role of Protozoa in Inactivation of Viruses in Water Pinheiro, M. D. O.; Power, M. E.; Butler, B. J. Applied and Environmental Microbiology, Vol. 73, Issue 2 https://doi.org/10.1128/AEM.02363-06	journal	November 2006
Effects of Graphene Oxide and Oxidized Carbon Nanotubes on the Cellular Division, Microstructure, Uptake, Oxidative Stress, and Metabolic Profiles Hu, Xiangang; Ouyang, Shaohu; Mu, Li Environmental Science & Technology, Vol. 49, Issue 18 https://doi.org/10.1021/acs.est.5b02102	journal	July 2015
Evaluation of toxicity of nanoclays and graphene oxide in vivo: a Paramecium caudatum study Kryuchkova, Marina; Danilushkina, Anna; Lvov, Yuri Environmental Science: Nano, Vol. 3, Issue 2 https://doi.org/10.1039/C5EN00201J	journal	January 2016
Interactions of multiwalled carbon nanotubes with algal cells: Quantification of association, visualization of uptake, and measurement of alterations in the composition of cells Rhiem, Stefan; Riding, Matthew J.; Baumgartner, Werner Environmental Pollution, Vol. 196 https://doi.org/10.1016/j.envpol.2014.11.011	journal	January 2015
Phase Distribution of ¹⁴ C-Labeled Multiwalled Carbon Nanotubes in Aqueous Systems Containing Model Solids: Peat Zhang, Liwen; Petersen, Elijah J.; Huang, Qingguo Environmental Science & Technology, Vol. 45, Issue 4 https://doi.org/10.1021/es1026097	journal	February 2011
Isolation of exosomes by differential centrifugation: Theoretical analysis of a commonly used protocol Livshits, Mikhail A.; Khomyakova, Elena; Evtushenko, Evgeniy G. Scientific Reports, Vol. 5, Issue 1 https://doi.org/10.1038/srep17319	journal	November 2015
Toxicity of ZnO and CuO nanoparticles to ciliated protozoa Tetrahymena thermophila Mortimer, Monika; Kasemets, Kaja; Kahru, Anne Toxicology, Vol. 269, Issue 2-3 https://doi.org/10.1016/j.tox.2009.07.007	journal	March 2010
NanoE-Tox: New and in-depth database concerning ecotoxicity of nanomaterials Juganson, Katre; Ivask, Angela; Blinova, Irina Beilstein Journal of Nanotechnology, Vol. 6 https://doi.org/10.3762/bjnano.6.183	journal	January 2015
Adaptive Interactions between Zinc Oxide Nanoparticles and Chlorella sp. Chen, Pengyu; Powell, Brian A.; Mortimer, Monika Environmental Science & Technology, Vol. 46, Issue 21 https://doi.org/10.1021/es303303g	journal	October 2012
Biomagnification of cadmium selenide quantum dots in a simple experimental microbial food chain Werlin, R.; Priester, J. H.; Mielke, R. E. Nature Nanotechnology, Vol. 6, Issue 1 https://doi.org/10.1038/nnano.2010.251	journal	December 2010
Nanomaterials in Agricultural Production: Benefits and Possible Threats? Hong, Jie; Peralta-Videa, Jose R.; Gardea-Torresdey, Jorge L. ACS Symposium Series https://doi.org/10.1021/bk-2013-1124.ch005	book	January 2013
Differential Growth of and Nanoscale TiO ₂ Accumulation in Tetrahymena thermophila by Direct Feeding versus Trophic Transfer from Pseudomonas aeruginosa Mielke, Randall E.; Priester, John H.; Werlin, Rebecca A. Applied and Environmental Microbiology, Vol. 79, Issue 18 https://doi.org/10.1128/aem.01680-13	journal	July 2013
Separation of small ciliate protozoa from bacteria by sucrose gradient centrifugation. Berk, S. G.; Guerry, P.; Colwell, R. R. Applied and Environmental Microbiology, Vol. 31, Issue 3 https://doi.org/10.1128/aem.31.3.450-452.1976	journal	January 1976
Assessing the Role of Pseudomonas aeruginosa Surface-Active Gene Expression in Hexadecane Biodegradation in Sand Holden, P. A.; LaMontagne, M. G.; Bruce, A. K. Applied and Environmental Microbiology, Vol. 68, Issue 5 https://doi.org/10.1128/aem.68.5.2509-2518.2002	journal	May 2002
Enhanced Exopolymer Production and Chromium Stabilization in Pseudomonas putida Unsaturated Biofilms Priester, J. H.; Olson, S. G.; Webb, S. M. Applied and Environmental Microbiology, Vol. 72, Issue 3 https://doi.org/10.1128/aem.72.3.1988-1996.2006	journal	March 2006

Cited By (3)

Challenges in characterizing the environmental fate and effects of carbon nanotubes and inorganic nanomaterials in aquatic systems Laux, Peter; Riebeling, Christian; Booth, Andy M. Environmental Science: Nano, Vol. 5, Issue 1 https://doi.org/10.1039/c7en00594f	journal	January 2018
Strategies for robust and accurate experimental approaches to quantify nanomaterial bioaccumulation across a broad range of organisms Petersen, Elijah J.; Mortimer, Monika; Burgess, Robert M. Environmental Science: Nano, Vol. 6, Issue 6 https://doi.org/10.1039/c8en01378k	journal	January 2019
Challenges in characterizing the environmental fate and effects of carbon nanotubes and inorganic nanomaterials in aquatic systems Laux, Peter; Riebeling, Christian; Booth, Andy M. RWTH Aachen University https://doi.org/10.18154/rwth-2018-220605	text	January 2018

Similar Records

Bioaccumulation of Multiwall Carbon Nanotubes in Tetrahymena thermophila by Direct Feeding or Trophic Transfer

Journal Article · Mon Jul 11 00:00:00 EDT 2016 · Environmental Science and Technology · OSTI ID:1377787

Mortimer, Monika; Petersen, Elijah J.; Buchholz, Bruce A.; +2 more

Importance of passive diffusion in the uptake of polychlorinated biphenyls by phagotrophic protozoa

Journal Article · Mon May 01 00:00:00 EDT 2000 · Applied and Environmental Microbiology · OSTI ID:1377787

Kujawinski, E B; Farrington, J W; Moffett, J W

Emerging investigator series: quantification of multiwall carbon nanotubes in plant tissues with spectroscopic analysis

Journal Article · Wed Dec 12 00:00:00 EST 2018 · Environmental Science: Nano · OSTI ID:1377787

Das, Kamol K.; Nava, Valeria; Chang, Che-Wei; +3 more

Related Subjects

77 NANOSCIENCE AND NANOTECHNOLOGY
54 ENVIRONMENTAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRONMENTAL POLLUTANT EFFECTS ON LIVING ORGANISMS AND BIOLOGICAL MATERIALS
59 BASIC BIOLOGICAL SCIENCES
37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
Pseudomonas aeruginosa
Tetrahymena thermophila
carbon-14
sucrose
iodixanol
bioaccumulation
bioconcentration
Stokes' law

Title: Separation of Bacteria, Protozoa and Carbon Nanotubes by Density Gradient Centrifugation

Citation Formats

References (44)

Cited By (3)

Similar Records

Related Subjects