Dirt in the Wound: Evaluating the Role of Iron in Antibacterial Minerals
Abstract
The recent discovery of antibacterial clay deposits which are effective in killing antibiotic resistant bacteria may lead to the discovery of mineral based antibacterial mechanisms. These antibacterial clays have been shown to prevent the growth of a broad spectrum of bacteria, including methicillin-resistant Staphylococcus aureus MRSA and extended-spectrum beta lactamase (ESBL) Escherichia coli (antibiotic resistant strains) when tested in vitro. This study investigates the first antibacterial mineral deposit identified in the United States, the Oregon Mineral Technologies (OMT) mine, which formed from the hydrothermal alteration of porphyry andesites. Our hypothesis is that mixed-layered clay minerals containing nano-iron sulfides can release soluble transition metals at low pH which are antibacterial due to the rapid influx and precipitation of intracellular metal-oxides while generating reactive oxygen species (ROS) and damaging bacterial membranes. To test this hypothesis, E. coli (ATCC 25922) was reacted with clay suspensions and clay leachates (solutions equilibrated with clays for 24 hrs). Inductively coupled plasma mass spectrometry (ICP-MS) was used to measure the soluble transition metals that are leaching from the clays. Bioimaging using scanning transmission electron microscopy-electron energy loss spectroscopy (STEM-EELS) and scanning transmission X-ray microscopy (STXM) were used to investigate the precipitation of intracellular mineral particles and redox state of the soluble metals reacting with the bacteria. Reactive oxygen species (ROS) were measured using a spectrophotometric hydrogen peroxide assay (H2O2) assay. Aldehydes were measured using HPLC-UV-Vis (high-performance liquid chromatography-ultraviolet-visible). Antibacterial susceptibility testing and ICP-MS elemental analysis of the leachates reveals that low pH (2.5-3.1) samples containing mM levels of soluble Fe, Al and Ca are antibacterial. All other potential toxins are below the minimum inhibitory concentrations for bactericide. The acidic pH is not the only factor contributing to the antibacterial effect. The intracellular particles observed upon cell death were determined to be Fe-oxides by STEM-EELS. STXM iron maps of single cells indicate that soluble Fe2+ and Fe3+ are adsorbing to the bacterial cell walls. The adsorption of reduced iron to the cell walls of bacteria can result in lipid peroxidation and the concurrent release of toxic aldehydes. Results from the HPLC-UV-Vis aldehyde assay reveal that the antibacterial leachates cause lipid peroxidation and the release of mono-aldehydes at μM levels from bacterial cell walls. The hydrogen peroxide and ferrous/ferric iron assay of the mineral leachates indicates that H2O2 is being generated in the presence of Fe2+, ultimately generating hydroxyl radicals which are toxic to bacterial lipids, proteins and DNA.
- Publication:
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AGU Fall Meeting Abstracts
- Pub Date:
- December 2013
- Bibcode:
- 2013AGUFM.B23E0600M
- Keywords:
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- 0461 BIOGEOSCIENCES Metals;
- 0463 BIOGEOSCIENCES Microbe/mineral interactions;
- 0448 BIOGEOSCIENCES Geomicrobiology;
- 0499 BIOGEOSCIENCES New fields (not classifiable under other headings)