On the mechanism of microbe inactivation by metallic iron 1 
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Chicgoua Noubactepa,b 
aAngewandte Geologie, Universität Göttingen, Goldschmidtstraße 3, D-37077,  Göttingen, Germany. 
bKultur und Nachhaltige Entwicklung CDD e.V., Postfach 1502, D-37005 Göttingen, Germany 
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e-mail: cnoubac@gwdg.de; Tel. +49 551 39 3191, Fax. +49 551 399379. 
 
Abstract 
This letter challenges the concept that the metallic iron (Fe0) surface contributes directly to 
the process of micro-organism inactivation in aqueous solutions. It is shown that any 
antimicrobial properties of Fe0 is related to the cycle of expansion/contraction accompanying 
aqueous iron corrosion. This demonstration corroborates the concept that aqueous 
contaminant removal in the presence of Fe0 mostly occurs at the Fe-oxide/water interface or 
within the oxide-film on Fe0. 
Keywords: Adsorption, Antimicrobial agent, Co-precipitation, Zerovalent iron. 
1 Introduction 
Following the successful use of micro-scale metallic iron (Fe0) for groundwater remediation 
[1-4], micro- and nano-Fe0 have shown promise as strong antimicrobial agents against a broad 
spectrum of bacteria and viruses [5-8]. While the efficiency of Fe0 for micro-organism 
inactivation is certain, the reported inactivation mechanisms are not convincing. The 
antimicrobial effect of Fe0 has been reported to involve the generation of intracellular 
oxidants (e.g. OH° and FeIV) produced by the reaction with hydrogen peroxide or other 
species, as well as a direct interaction of Fe0 with cell membrane components [8]. However, it 
is clear that this elucidation has not properly considered three important facts: (i) at pH > 5.0 
the surface of Fe0 is permanently covered by an oxide-film and is therefore not directly 
accessible to microbes [9], (ii) oxide-film components (Fe-oxides) are antimicrobial agents 
and might independently inactivate microbes [10,11], and more importantly (iii) Fe0 oxidation 
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coupled to Fe-oxide precipitation and oxide-film formation is a dynamic process [9]. 
Accordingly, Fe-oxides are continually produced for micro-organism inactivation, ideally 
until Fe
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0 is totally depleted. Addition of Fe0 as a remediation strategy is therefore appealing 
due to the progressive slow release of highly reactive Fe-oxides. In contrast, in systems using 
less sustainable synthetic Fe-oxides (also as coatings on granular surfaces) the initial 
inactivation capacity may be high but the retention capacity is limited and microbe 
inactivation is due to pure adsorption and/or mechanical trapping. 
The inactivation of pathogens in filtration systems is known to occur through adsorption, 
mechanical trapping (size-exclusion or straining), natural death, and predation [12,13]. 
Pathogen predation is not addressed in this work. Natural death mostly results from transport 
retardation through straining or adsorption and died microbes may be transported across the 
filter. Adsorption results from electrostatic interactions between pathogens and involved solid 
phases. For example, in comparison to bacteria inactivation, slow sand filters have shown 
limited inactivation efficiency for viruses in natural waters (6.0 ≤ pH ≤ 9.0) [12]. This 
observation was attributed to the fact that, under these pH conditions, sand and most viruses 
are negatively charged, leading a net repulsion and thus relative less virus removal by sand 
filtration [12,14]. Fe-oxide amended sand filters have shown improved pathogen inactivation 
[5,12,15] because the positively charged surface of oxide layer may electrostatically adsorb 
viruses [16]. Fe-oxides are either immobilized on granular media (e.g. [15]) or added as 
granular Fe0 (e.g. [12]) or supported nano-Fe0 [17]. 
Investigations regarding the addition of Fe0 to slow sand filter for safe drinking water 
provision at household level have boomed in recent years [5,8,12].  The next section give an 
overview of efforts to elucidate the mechanism of microbe inactivation. 
1.1 Apparent quest for the mechanism of microbe inactivation 
The presentation above has shown that the scientific community is still looking for plausible 
explication of the efficiency of Fe0 for the inactivation of microbes [8,18-20]. For example, 
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Kim et al. [8] reported on the elucidation of the removal mechanism of MS2 coliphage (a 
virus) by Fe
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II and nano-Fe0 and suggested the need of more research to characterize the 
impact of nano-Fe0 on other microbes (e.g. bacterial species, viruses, protozoan cysts, and 
complex matrices). Clearly, the mechanism of micro-organism inactivation is considered 
species-dependant. This approach is the one that has been used for chemical contamination 
but has been proven superfluous because contaminant removal in Fe0/H2O systems is not 
primarily a characteristic of any contaminant, but a characteristic of aqueous iron oxidation at 
pH > 5.0 [9,21,22]. Before recalling, the mechanism of contaminant removal by Fe0, the 
following conclusion of Kim et al. [8] should be given: “The applications of nano-Fe0 to 
inactivate viruses could be broader than for bacteria because nano-Fe0 maintains virucidal 
activity in both the presence and absence of oxygen, whereas aerobic conditions may limit the 
bactericidal activity of nano-Fe0.” 
2 Mechanism of aqueous contaminant removal by Fe0
The suitability of Fe0 as a universal material for safe drinking water production has been 
theoretically discussed during the past three years [23-28]. The basic idea is that iron 
(hydr)oxides are good adsorbents of chemical and microbial contaminants. This idea has 
already led to the development of metal hydroxide-coated granular materials (e.g. gravel, 
sand) as an efficient adsorption medium in water treatment [11,29]. The approach of using Fe0 
as in-situ iron oxide generator for contaminant removal was also known but was tested on a 
case-by-case basis for selected contaminants: e.g. arsenic [30,31] and viruses [5,8]. However, 
filters designed for As removal [30] were able to remove more that 27 other species including 
heavy metals, organics compounds and pathogens [32-34]. This latter observation clearly 
exceeded design expectations and demand for accurate explanations. 
The observed efficiency of Fe0-based filters was explained by considering the dynamic nature 
of Fe0 corrosion within the porous media (filters) [23,24,28]. In fact, iron corrosion is 
volumetric expansive in nature [35,36]. Depending on the oxygen availability, the volume of 
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formed iron oxides may be up to 6.40 times larger that the volume of Fe0 in the metal lattice 
[35]. However, formed oxides go through intermediate stages of more voluminous hydroxides 
which are colloidal in nature and very adsorptive for any dissolved species. Accordingly, the 
process of Fe
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0 corrosion is a cycle of expansion and contraction events. Expansion 
corresponds to the transformation “Fe0 ⇒ voluminous hydroxides”. Contraction corresponds 
to the transformation “voluminous hydroxides ⇒ final oxides”. During these events, 
contaminants are basically enmeshed in the mass of precipitating oxides (within the oxide-
film). Additionally, contaminant adsorption onto the surface of resulted precipitates is also 
efficient (at the interface Fe-oxide/H2O). The overall process in packed beds was termed as 
“reactive filtration” and convincingly explained the reported efficiency of Fe0-based filters 
[24,28]. 
It should be explicitly stated that the extent and “apparent mechanism” of contaminant 
removal in laboratory experiments (including microbe inactivation) depend on the used 
experimental designs. This issue will not be further discussed here. However, it should be 
stated that to be relevant for practical situations, Fe0 materials should be tested under 
conditions in which the formation of oxide-films at their surface is not disturbed. The 
initiation and growth of oxide-films are highly dependent on the Fe0 intrinsic reactivity and 
availability of reactants [37,38]. It is well-documented that after the formation of the oxide-
film on the Fe0 surface, the Fe0 oxidation progressed at a significantly reduced speed. This 
phase of reduced oxidation kinetics corresponds to real-world situations for Fe0-based filters. 
Accordingly, long-term laboratory experiments are suitable for a better understanding of the 
operating mode of Fe0 filters. 
The use of Fe0 for microbe inactivation was first tested on a pragmatic basis based on the 
success of Fe0 in permeable reactive barriers [5]. A science-based introduction of elemental 
metal for microbe inactivation was reported earlier [39] as discussed in the next section. 
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2.1 Elemental metals for microbe inactivation 104 
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Irrespective from the author’s previous works on the Fe0/H2O system summarized in refs. 
[24,28], the mechanism of microbe removal by Fe0 can be derived by analogy to the process 
of electrocoagulation (EC) using Al and Fe as sacrificial anodes. The effects of Al0/Fe0 are 
based on spontaneous dissolution in contact with water, with generation of AlIII/FeIII-species 
and OH- ions, and finally voluminous insoluble Al(OH)3/Fe(OH)3. If O2 is absent or limited, 
less voluminous Fe(OH)2 will be formed. That is the sole difference between oxic and anoxic 
conditions. Bearing in mind the great efficiency of Al0 EC and Fe0 EC for the aqueous 
removal of many chemical pollutants, efficient inactivation of microbiological water pollutant 
is expected too. This principle was used by Bojic et al. [39] to develop a very efficient micro-
alloyed Al0-based composite for water treatment. It should be recalled that conventional Al0 is 
very low reactive as it is instantaneously covered by an impervious film Al2O3 film on Al0 
[40]. The same trend is observed for elemental zinc. From a pure thermodynamic perspective, 
however, Al and Zn are stronger reducing agents than Fe0 [40]. The standard electrode 
potentials for the redox couples of the three elements are: -1.66 V for AlIII/Al0, -0.763 V for 
ZnII/Zn0, -0.44 V for FeII/Fe0 and 0.77 V for FeIII/FeII [40, 41]. 
Considering the thermodynamics of oxide-film formation on the three metals (Al, Fe and Zn), 
it appears that Fe0 is the sole multivalent element (FeII, FeIII) [41]. Because of differences in 
size and chemical properties of  Fe0, FeII and FeIII species, the formation of an impervious 
oxide-film on Fe0 is not likely, this is the rational for the better suitability of Fe0 for 
environmental remediation.  In other words, to render Al and Zn (and other aqueous reactive 
metals) suitable for environmental remediation, tools have to be found to avoid the formation 
of an impervious oxide-film on their surface. Despite this evidence, researchers are continuing 
to discuss the suitability of conventional Zn0 for aqueous contaminant removal [42-48]. For 
Al0, Bojic et al. [39,49-51] have presented an efficient micro-alloyed composite. The 
composite consists of micro-alloyed aluminium coated over a thin iron net. 
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This microalloyed Al0-based composite has been successfully used for the aqueous removal 
of trihalomethanes, textile dyes, natural organic mater, pesticides, heavy metals and 
Escherichia coli [49-51]. The removal mechanisms had been reported in terms of flocks of 
aluminium hydroxide acting “as adsorbents and/or traps for ions, molecules or suspended 
particles thus removing them from the solution by sorption, co-precipitation or electrostatic 
attraction followed by coagulation” [51]. This description corresponds to adsorption and co-
precipitation as a fundamental mechanism of chemical contaminant removal and micro-
organism inactivation [9,52]. It should be recalled that generated iron species will not 
segregate bacteria, chemical contaminants and viruses. All begin or pathogenic species are 
removed from the aqueous phase, provided that enough time is left for sufficient production 
of removing agents. Accordingly, all reports on the demonstration of microbe-specific 
interactions leading to other removal mechanisms (e.g. cytotoxic) were somehow faulty. 
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3 Concluding remarks 
In a recent review on biological research, Brenner [53] stated that the conversion of data into 
knowledge constitutes a great challenge for future research. It is intuitive to conclude that 
this conversion will be very difficult when the data are produced on a pragmatic basis. This 
has been the case for the use of Fe0 for water treatment [8,18,19]. Ideally concepts (e.g. 
theories of the system) should exist which are to be approved or disproved by experimental 
data. In the absence of any concept, there is no guide to constrain the choice of model. In 
addition, most of the observations (e.g. nature of corrosion products, percent removal) made 
by individual researchers are static snap-shots and their measurements could be 
experimentally impacted [53]. Therefore, it will be impossible to use available data to 
understand the dynamic processes of contaminant removal by aqueous iron corrosion. The 
situation is exacerbated by the huge difference of time scales between laboratory experiments 
(hours to days) and field application (years). 
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For a more effective development of the iron reactive wall technology, the state-of-the-art 
knowledge on the mechanism of contaminant removal should be considered by all 
investigators regardless the size of used materials (nm, μm and mm) and the nature of the 
contaminant (biological, chemical or physical). The sole impact of the particle size is on the 
kinetic aspects [2,54]. Factors introducing biases in the experimental protocols have been 
intensively discussed [55]. These factors included [55]: the available reactive sites (Fe
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particle size, Fe0 loading), the intrinsic reactivity of used Fe0 [56], the volume of the solution, 
the contaminant concentration, the mixing type (shaking, stirring, vortex), the mixing 
intensity, and the Fe0 pre-treatment for batch experiments. For column experiments, factors 
influencing the treatment efficiency include: the reactivity of used Fe0, the proportion of Fe0 
in the mixture, the nature of the admixing agent (e.g. gravel, perlite, pumice, sand), the water 
characteristics (dissolved O2 level, pH value, nature of contaminants), and the hydraulic loads. 
In conclusion, the long-lasting debate on the toxixity or the mode of toxixity of Fe0 [3,57,58] 
should be re-oriented. Whether Fe0, (nano-Fe0), released FeII, generated HO° and FeIV are 
cytotoxic or not, quantitative microbe removal is likely. Nano-Fe0 reacts and depletes rapidly 
[2,54], producing more Fe-oxides per time unit. Moreover, it is difficult to understand the 
relevance of Fe0, FeIV and HO° as virucidal and bactericidal agents [8] when microbes are 
readily and irreversibly removed from the aqueous phase by the dynamic process of iron 
corrosion. 
Acknowledgments 
Thoughtful comments provided by Angelika Schöner (FSU Jena - Germany) on the revised 
manuscript are gratefully acknowledged. The manuscript was improved by the insightful 
comments of anonymous reviewers from the Journal of Hazardous Materials. 
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