International Journal of Biotechnology and Bioengineering Research.

ISSN 2231-1238, Volume 4, Number 3 (2013), pp. 191-196

Research India Publications
http://www.ripublication.com/ ijbbr.htm

Engineered Nanomaterials as Disinfectants:

Benefits and Health Concerns
A. Godara1, S. Srivastava2, A. Anandan3, B. Dayal4, and A. Kumar5*
1

Department of Biochemical and Bioengineering,

Indian Institute of Technology Delhi, New Delhi.
2
Department of Civil Engineering, Indian Institute of Technology Delhi, New Delhi.

Abstract
Engineered nanomaterials (ENMs) such as Ag, TiO2, ZnO, etc. are
currently being used as disinfectants for killing pathogens in water
treatment plants. However, this increased usage also raises an
important question of their long-term effects on pathogens defense
systems. This study aims to understand toxic effects of ENMs on
pathogens and identify current research needs for developing better
understanding. First bacterial disinfection with ZnO NPs was
considered and toxic effects were correlated with bacterial defense
mechanisms. Literature reports and findings were synthesized to
identify issues, needing immediate research and regulatory attention.
Analysis indicated that ENMs are better disinfectants when used in
combination with antibiotics by releasing oxygen radicals and inducing
oxidative stress. Toxicity of these effects depends on ENM dose level,
exposure duration, and bacterial defense system. Bacteria employ
defense mechanisms such as release of extracellular polymeric
substance which can reduce toxic effects of ENMs. Experience with
exposure of bacteria to antibiotics indicates that bacteria develop
resistance mechanisms by adapting themselves against antibiotics by
employing phenotypic and genetic changes. This kind of information is
currently not available for exposures of bacteria to ENMs, which
leaves the question of long-term effect of ENMs on bacteria
unanswered. Overall, it is still unclear how bacteria would react to
exposures of ENMs after repetitive exposures and what conditions are
conducive for them to develop resistance against ENMs. These
uncertainties in bacterial defense mechanisms towards ENMs have
implications on effectiveness of ENMs as disinfectants and may pose

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health concerns, if ENMs are not effective to bacteria. Above
mentioned issues indicate the need for developing understanding of
development of pathogenic defense mechanisms against toxic effects
of ENMs, so that ENMs could be used effectively without posing
health risks.
Keywords: Disinfections; Human health; Nanomaterials; Pathogens;
Resistance.

1. Introduction
Engineered nanomaterials (ENMs) such as Ag, TiO2, ZnO, etc. are currently being
used as disinfectants for killing pathogens in water treatment plants. However, this
increased usage also raises an important question of their long-term effects on
pathogens defense systems. Ineffectiveness of ENMs in killing bacteria could result in
human exposures of bacteria and thus increasing chances of microbial exposures from
wastewater effluent or river water. This study aims to understand toxic effects of
ENMs on pathogens and identify current research needs for developing better
understanding.

2. Methodology
First, bacterial disinfection with Ag and ZnO NPs was considered and toxic effects
were correlated with bacterial defense mechanisms. Literature reports and findings
were synthesized to identify issues, needing immediate research and regulatory
attention.

3. Results and Discussion

3.1. ZnO as antimicrobials
Nano particles such as Zinc Oxide were reported to possess antibacterial properties
against Bacillus subtilisand Escherichia coli. ZnO NP toxicity to bacteria depends on
NP size as well as concentration. The interaction of these two factors determines extent
of ZnO NP toxicity to a given type of bacteria. This results in large variability in ZnO
disinfection data and its killing dose, i.e., IC50s (growth inhibition) for different
bacteria ranges from <1 mg/L to several hundred mg/L or higher. The variability also
exists among a same E.colispecies. for example, ZnO NPs with particle size (10
30 nm) suspended in bacterial growth media had a minimum inhibition concentration
(MIC) about 500 mg/L for wild-type E. coli (Premanathan et al., 2011). The ZnO size
also affects extents of toxicity. Particles aggregate in suspension and the resultant size
determines toxicity to bacteria (Adams et al., 2006).
Overall, toxicity studies using ZnO NPs need to consider combined interaction of
these two factors for given bacterial specie for understanding disinfection

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effectiveness. ZnO NP may distort and damage the membrane wall of the bacteria,
resulting in a leakage of intracellular contents and eventually the death of bacterial
cells.Also ZnO NPs have strong affinity for sulphur containing bases in cell
membranes and phosphorus containing macromolecules thus making cell walls and
DNA preferential sites bind to Nano particles which leads to adverse cell effects, our
study here mainly focuses on interaction of ZnO nanoparticles and E. Coli bacteria.
Another possible mechanism for ZnO antibacterial activity is the release of Zn2+ ions
due to presence of H2O2 species (Domnech and Prieto, 1986). However, there is still
a current lack of knowledge on interaction of NPs with the bacterial cell wall and
possible permeation of the NPs into the bacterial cells (Jiang et al. 2009).
3.2. Toxicity mechanisms
In general, NPs release oxygen radicals which interact with cell wall and damage them
(DNA damage, lactate dehydrogenase leakage, GSH depletion, and disruption of
membrane size) (Hussainet al. 2005; Jeng and Swanson 2006; Chen et al.2007;
Brayneret al. 2006). NPs generate these species which depends on NP surface area
(and thus on particle size) (Ahmed et al.2010), indicating the role of NP size in
affecting NP-associated toxicities. Further, analysis indicated that NPs are better
disinfectants when used in combination with antibiotics by releasing oxygen radicals
and inducing oxidative stress. Toxicity of these effects depends on ENM dose level,
exposure duration, and bacterial defense system. Bacteria employ defense
mechanisms, such as release of extracellular polymeric substance which can reduce
toxic effects of NPs. Experience with exposure of bacteria to antibiotics indicates that
bacteria develop resistance mechanisms by adapting themselves against antibiotics by
employing phenotypic and genetic changes. This kind of information is currently not
available for exposures of bacteria to NPs, which leaves the question of long-term
effect of NPs on bacteria unanswered.
3.3. Bacterial defense mechanisms
In response to stress, bacteria employ different defence mechanisms to reduce extent of
stress and toxicity. One of the defence mechanisms is production of Extracellular
polymeric substances (EPS) which binds with external contaminants and reduces its
availability to bacteria and thus effective toxic concentration available for affecting
bacteria is reduced. These EPS are produced due to metabolic activities in bacteria and
is secreted outside the cell well. It forms a protective layer for the cells against the
harsh external environment such as NP exposure, biocides, and sudden changes in pH,
absorbing exogenous nutrients and organic molecules and thus protects the bacteria.
Another method of bacterial defence is resistance development in bacteria due to
change in genetic material. This happens due to bacterial adaptation of external
environment and thus it reduces overall impact of external stressors. This information
leaves the question whether bacteria develop resistance to NPs during long-term
exposure. Review indicates lack of data on development of bacterial resistance to NPs
during interaction of bacteria with NPs. this might be possible however this needs to be

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investigated to finally determine extent of resistance development, if any and upto

what extent and in what conditions? Nanoparticles as well as antibiotics impart stress
to the cell wall of bacteria. So there is a possibility that bacteria might develop
resistance to nanoparticles like in case of antibiotics.
3.4. Exploring analogy between bacterial defense mechanisms against antibiotics
and nanoparticles
An idea about bacterial resistance to NPs could plausibly be drawn by taking analogy
with bacterial resistance to antibiotics. Resistance mechanisms include (1) inactivation
of the compound by detoxifying enzymes, (2)reduced cell permeability or expulsion of
the drug by specific or non-specific pumps, and (3) modification of the antibiotic
targets. Bacteria can also acquire a resistance phenotype via global or specific
transcriptional regulators. It is important to know if bacteria employ these resistance
mechanisms when it is exposed to NPs and if yes, up to what extent. This information
need to be obtained to understand bacterial defence mechanisms against NPs and
finally development of resistance mechanisms in long-term exposure.

4. Summary and Conclusions

Overall, it is still unclear how bacteria would react to exposures of NPs after repetitive
exposures and what conditions are conducive for them to develop resistance against
NPs. These uncertainties in bacterial defense mechanisms towards NPs have
implications on effectiveness of NPs as disinfectants and may pose health concerns, if
NPs are not effective to bacteria. Above mentioned issues indicate the need for
developing understanding of development of pathogenic defense mechanisms against
toxic effects of NPs, so that NPs could be used effectively without posing health risks.

5. Acknowledgement
This work was supported partly by the IIT Delhi, partly through the Department of
Science and Technology (India) grant (No. DST/TM/WTI/2K11/301). Views
presented here do not represent views of the agency and it solely represents views of
authors.

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