Journal of Environmental Science and Health Part A (2009) 44, 1485–1495

Copyright
C Taylor & Francis Group, LLC

ISSN: 1093-4529 (Print); 1532-4117 (Online)

DOI: 10.1080/10934520903263231

Aggregation and toxicity of titanium dioxide nanoparticles

in aquatic environment—A Review
VIRENDER K. SHARMA
Chemistry Department, Florida Institute of Technology, Melbourne, Florida, USA

The use of nanoparticles—particles with size ∼1–100 nm is increasing worldwide. This is particularly the case for applications
of titanium dioxide nanoparticles (nano-TiO2 ) in consumer products, which have expanded at a fast rate in the last decade. The
properties of nano-TiO2 differ significantly from bulk-TiO2 of the same composition because of an increase in surface area. A release
of nano-TiO2 from application sources to the aquatic environment may pose possible risks due to their bioavailability and toxicity.
The aggregation of nano-TiO2 plays an important role in the environmental effects of nanoparticles because the size and shape of
nanoparticles will determine the magnitude of any potentially toxic effect. Aggregation is affected by pH, ionic strength, and ionic
identity (inorganic and organic) of aqueous suspensions and is reviewed in this paper. The current information on the evaluation of
ecotoxicological hazards of nano-TiO2 to bacteria, algae, invertebrates, nematodes, and rainbow trout is also given.
Keywords: TiO2 , aggregation, ph, ionic strength, organic matter, aquatic toxicity, reactive oxygen species, bacteria, invertebrates,
algae.

Introduction
Nanoparticles are usually defined as particles smaller than
100 nm in at least two dimensions. The physical, chemi-
cal, and biological properties of nanoparticles are gener-
ally different from the properties of the same materials at
a larger scale. Nanomaterials have been shown to offer sig-
nificant potential applications for providing solutions to
technological and environmental challenges in fields such
as medicine, solar energy conversion, catalysis, and water
purification.[1–6] Because of such diverse applications of
nanomaterials in a wide range of industries, the production
of engineered nanomaterials is expected to increase from
400 tons to 58,000 tons in 2011–2020.[7] In other words, the
nanomaterials industry is growing with an estimated enter-
prise of $1 trillion by 2015.[8] With this growth, synthesis of
nanomaterials requires the use of the 12 principles of green
chemistry in order to eliminate hazards. Figure 1 demon-
strates the applications of these principles to nanoscience,
which suggest the need of developing new synthetic and
characterization tools.[9] A better understanding of bio/eco
techniques may also be needed to design safe novel mate-
rials efficiently.[9] Furthermore, the interaction of nanoma-
Address correspondence to Virender K. Sharma, Chem-
istry Department, Florida Institute of Technology, 150 West
University Boulevard, Melbourne, Florida 32901, USA. E-mail:
vsharma@fit.edu
Received June 25, 2009.
terials with DNA, lipids, cells, tissues, proteins, and bio-
logical fluids needs to be considered for evaluation of their
safety to humans and to the environment.[10–16] Moreover,
nanoparticles in aquatic environments may influence envi-
ronmental processes differently than the same materials of
larger size.[17] As shown in Figure 2, the reactivity of the
particle does not always increase with decreasing particle
size, but depends on the chemical reaction and the material
involved.[17] Four interrelated factors have been suggested
to cause changes in the reactivity at the nanoscale. These
are: (i) the effect on the thermodynamics of chemical reac-
tivity through the dependence of the surface free energy on
the particle size; (ii) increase in surface area as nanoparticles
get smaller and smaller; (iii) the variation in atomic struc-
ture in terms of bond angles, bond lengths, and vacancies
and other defects near and on surfaces; and (iv) modifi-
cation of electronic structure with size. The material and
the size range of the particles ultimately determine whether
one or all four of the factors contribute to size-dependent
chemical reactivity.[17]
Of the various nanomaterials, titanium dioxide (TiO2 )
has been of great interest. There are natural and man made
sources of TiO2 in the environment.[18–21] Natural sources
of TiO2 are dominated by the minerals ilmenite (FeTiO3 )
and rutile (TiO2 ), which are commonly found in platonic
and metamorphic rocks and also found as distinct minerals
in beach sands.[18] Because of the use of nano-TiO2 in paints
and coatings as self-cleaning, antimicrobial, and antifoul-
ing agents and in cosmetics as a UV-absorber, it is produced
1486 Sharma

Fig. 1. Translating the 12 green chemistry principles for application in the practice of green nanoscience. The principles are listed,
in abbreviated form, along with the general approaches to designing greener nanomaterials and nanomaterial production methods
and specific examples of how these approaches are being implemented in green nanoscience. Within the figure, PX, where X = 1−12,
indicates the applicable green chemistry principle. (Reproduced from [9] with the permission of the American Chemical Society)

in a large scale.[19–22] Although natural nano-TiO2 has been
found in river water,[23] recently, direct evidence of the re-
lease of synthetic nano-TiO2 from urban applications into
the aquatic environment has been documented.[24] It was
shown that nano-TiO2 leached out from the nano-TiO2
containing painted house facades and entered into receiv-
ing waters; giving concentrations in the range of a few µg
L−1 in a small stream.[24]
Model calculations on the release of nano-TiO2 into the
environment have been done using substance flow analy-
sis from products to air, soil, and water in Switzerland.[25]
The predicted environmental concentrations (PEC) un-
der a realistic scenario (RE) and high emission scenario
(HE) are given in Table 1. The PEC values for nano-
TiO2 in water ranged from 0.7–16 µg L−1 , which is nearly
equal to or higher than the predicted no effect concentra-
tion (PNEC) of <1 µg L−1 . These values gave risk quo-
tients (PEC/PNEC) of nano-TiO2 as >0.73 µg L−1 and
>15.83 µg L−1 in the RE and HE scenarios, respectively.
This indicates a possible ecotoxicity of nano-TiO2 in wa-
ter. It is clear that the behavior of nano-TiO2 needs to be
studied in the aquatic environment.
The toxicity of nanoparticles depends on both the
size and surface chemistry of nanoparticles in water.[26,27]
Aggregation of nanoparticles thus becomes critical in un-
derstanding toxicity, because of the relationship between
aggregation state, size, and observed effects.[28–31] This pa-
per therefore first reviews the aggregation of nano-TiO2
under natural conditions, followed by a discussion on tox-
icity of nano-TiO2 in the aquatic environment.

Table 1. Predicted environmental concentrations (PEC) of nano-

TiO2 in air, water, and soil. (Data were taken from[25] ).

Unit RE HE

Air µg m−3 1.5×10−3 4.2×10−2

Fig. 2. Generalized trend for size-dependent reactivity change of a Water µg L−1 0.7 16
material as the particle transition from macroscopic (bulk-like) to Soil µg kg−1 0.4 4.8
atomic. (Reproduced from [17] with the permission of the Royal
Chemical Society) RE: realistic scenario; HE: high emission scenario.
TiO2 nanoparticles in aquatic environment 1487

Fig. 3. Modification of nanoparticles in the environment. (Reproduced from [32] with the permission of Springer)

Aggregation It is therefore expected that the mobility of nano-TiO2

in natural environments may be dependent on the size of
Free nanoparticles usually tend to aggregate in the envi- nanoparticles. Aggregation of nano-TiO2 may limit trans-
ronment and then can possibly be eliminated or trapped port due to settling out of solution. Besides pH, ionic
through sedimentation (Fig. 3). Generally, aggregated strength and the nature of the electrolyte(s) in aqueous
nanoparticles are less mobile and can interact with filter suspensions influence the aggregation of nano-TiO2 .[40] At
feeders and sediment-dwelling animals.[32] Agglomeration pH ∼4.5 and in 0.0045 M NaCl suspensions, 4–5 nm TiO2
is thus important in understanding the fate and transport readily aggregated to average diameter sized particles of 50–
of nanoparticles in the natural environment. Though stud- 60 nm. If the ionic strength increased to 0.0165 M NaCl,
ies have been published on the transport of small par- the formation of micro-sized aggregates occurred within 15
ticles, experiments often involve particles of an organic minutes. In increasing the pH to 5.8–8.2, the formation of
nature, such as polystryrene beads, fullerols, and carbon
nanotubes.[33–36] Limited information on the transport of
nano-TiO2 is available.[37,38] The pH of experiments was
found to be an important parameter in the aggregation of
nano-TiO2 .[37]
Figure 4 shows the effect of pH on aggregate size over the
pH range of 1–12. As the pH approached the point of zero
charge (pHzpc ), the aggregation size increased. This trend
is similar to phenomena observed in the settling of parti-
cles. It seems that electrorepulsion is fundamental in the
understanding of the conditions under which aggregation
of nanoparticles occurs. As the solution pH approaches
the pHzpc , the repulsion between nanoparticles of similar
surface potentials decreases, making it easier for particles
to aggregate (Fig. 4). Research has shown that more than
80% of suspended particles and aggregates can be mobilized
over the pH range of 1–12, except when close to pHzpc .[37]
It has been shown independently that the pHzpc of nano- Fig. 4. Particle/aggregate size populations as a function of pH
TiO2 changes with size. The smallest particle sizes show the as measured by dynamic light scattering (DLS). The bars repre-
lowest pHzpc (pHzpc (3.6 nm) = 4.8) while largest particles sent the range. Reproduced from [37] with the permission of the
have a higher pHzpc (pHzpc (8.1 nm) = 6.2).[39] American Chemical Society)
1488
micro-sized aggregates happened within 5 minutes at even
low ionic strength of 0.0084–0.0099 M NaCl.
Importantly, a significantly faster aggregation of nano-
TiO2 took place in replacing NaCl with CaCl2 aqueous
suspensions at an ionic strength of 0.0128 M at pH 4.8.[40]
The results suggest that cations present in soils and surface
waters may play an important role in the aggregation of
nano-TiO2 .
The studies mentioned above do not account for the in-
fluence of the presence of natural organic matter (NOM)
such as microbial and humic substances, which are present
in the natural environment. In the environment, nanopar-
ticles can adsorb NOM to form complexes (or NOM coat-
ings) which may cause particles to acquire negative surface
charge. This alters the fate and transport of nanoparti-
cles in the aquatic environment.[41,42] This may also alter
the sorption of toxic hydrophobic organic compounds by
NOM.[43–48]
Humic substances can significantly influence the trans-
port of nanoparticles by modifying electric charge, size,
and the chemical nature of the particles.[49] Recent work
on the aggregation of nano-TiO2 in the presence of nat-
ural organic acids, showed such influences on the stabil-
ity of TiO2 suspensions.[50] The presence of oxalic and
adipic acids destabilized suspensions.[50] However, experi-
ments were performed at pH values far from the pHzpc and
may not realistically represent natural conditions found in
the environment. Only recently, the effects of humic sub-
stances on the aggregation of nano-TiO2 were studied by
using Suwannee River Fulvic Acid (SRFA) under various
physicochemical conditions.[51]
Diffusion coefficients and weight average diameters of
TiO2 were determined as a function of pH (2–8), ionic
strength (0.005–0.1 M), and SRFE (0.5–2.0 mg L−1 ). An
increase in ionic strength usually resulted in increased ag-
gregation at a given pH. Interestingly, adsorption of the
SRFA onto particles, caused less aggregation of nano-TiO2 ,
perhaps due to increased steric repulsion. This is in contrast
to earlier experiments performed with oxalic acid.[50] The
influence of the concentration of SRFA on the aggrega-
tion of nano-TiO2 is shown in Figure 5. An increase in
the concentration of SRFA increased disaggregation at all
studied pH values. At pH 8.0, which is above the pHzpc , col-
loidal suspensions were less disaggregated in the presence
of 1.0 mg L−1 SRFA than its absence. At concentration
of 2.0 mg L−1 SRFA, increasing steric stabilization of the
particles likely contributed to a decrease in hydrodynamic
diameters (see Fig. 4). Results of the study[51] suggest that
the dispersion and mobility of nano-TiO2 may occur to a
greater extent than predicted from laboratory experiments.
Recently,transport properties of anatase polymorph
TiO2 nanoparticles (ANTNPs) were evaluated in clean
silica, amorphous aluminum, and iron hydroxide-coated
sands.[52] In this study, the nanoparticles were also encapsu-
lated by carboxymethyl cellulose (CMC) and were referred
to as CMC-ANTNPs. CMC is used in several industries
Sharma
Fig. 5. Variation of diffusion coefficients and weight average di-
ameters of 1.0 mg L−1 of TiO2 nanoparticles as a function of the
concentration of SRFA for an ionic strength of 0.01 M and for
three different pH values: 2.0 (
), 5.5 (O) and 8.0 (). The arrow
indicates the size of the disaggregated TiO2 nanoparticles as pro-
vided by NanoAmor. (Reproduced from [51] with the permission
of the American Chemical Society)
including paper and detergents, paints, ceramics, glues and
adhesives and a release of such compounds may influence
the mobility of TiO2 nanoparticles in the aquatic environ-
ment. Initially, influence of CMC on the surface charge
and particle size of ANTNPs was studied (Fig. 6A). The
isoelectric point (IEP) of the particles decrease with the
addition of CMC. This reduction in the IEP creates steric
and double layer repulsions between particles and collector
surfaces to restrict attachment.[52]
The influence of pH on the measured electrophoretic
mobility (EPM) is also presented in Figure 5A. The EPM
decreased with increases in pH for both ANTNPs and
CMC-ANTNPs. The magnitude of EPM also decreased
with increase in the concentrations of Na+ and Ca2+ ions
(Fig. 6B). Bivalent Ca2+ had a greater effect than Na+ .
This cationic influence may be associated with a reduc-
tion of surface charge due to complexation with nega-
tively charged groups (e.g. COO− and OH− ) of CMC-
ANTNPs.[53] Interestingly, the hydrodynamic diameter in
the presence of Ca2+ increased from 228 to 423 nm while the
addition of Na+ decreased the diameter from 228 to 147 nm
(Fig. 6B).
This indicates that Ca2+ strongly suppresses the electric
double layer and the van der Waals attractive forces be-
came predominant. The addition of Ca2+ ions may also
result in aggregation of ANTNPs; similar to the forma-
tion of large-size aggregates of fullerene nanoparticles with
increased concentration of Ca2+ ions.[54] Comparatively,
the hydration shell effect of Na+ may result in the smaller
hydrodynamic diameter at high salt concentrations.[55]
Overall, results suggest that high molecular weight com-
pounds such as CMS influence the mobilization of
ANTNPs. TiO2 nanoparticles may thus enter into natu-
ral and engineered water resources more easily when CMC
encapsulated.[52]
TiO2 nanoparticles in aquatic environment 1489

Fig. 6. Measured electrophoretic mobility (EPM) and particle size hydrodynamic diameter (nm) of ANTNPs: (A) in the absence and
presence of CMC over pHs [ANTNPs concentration: 20 mg L−1 , CMC: 0.2%, background electrolyte: 1 mM NaCl; (B) as a function
of ionic strength and cation type: pH 6.4 ± 0.4, ANTNPs: 20 mg L−1 , CMC: 0.2%. (Reproduced from [52] with the permission of
the American Chemical Society).

Toxicity etration of nanoparticles into the skin.[57,58] However, it is

Mechanism
Because the size of nanoparticles is similar to that of typical
cellular components and proteins, nanoparticles can travel
inside the human body.[56] Some studies have shown pen-
important to know whether TiO2 particles are anatase and
rutile or monodispersed or aggregated in order to under-
stand their dermal toxicological impact.[59] TiO2 nanopar-
ticles can cause DNA and pulmonary damages.[60] In ex-
posing TiO2 nanoparticles to human endothelial cells, an
1490 Sharma

increase in the levels of IL-8 was observed.[61] Evidence is

accumulating that nanoparticles can induce the production
of reactive oxygen species (ROS), oxidative stress (OS), and
inflation in the vasculature and lungs.[5,62] During OS, the
production of ROS overwhelms the antioxidant defense ca-
pacity of the cell, causing adverse biological consequences.
It is also known that OS is the major source of
ROS, which included the superoxide anion (O·− 2 ), hy-
drogen peroxide (H2 O2 ), hydroxyl radicals (· OH), and
peroxynitrites.[63]
This would result in cell death due to oxidative DNA
damage and increases in the level of cellular nitric
oxide.[56,64,65] Several in vitro studies have reported that
nano-TiO2 caused OS mediated toxicity in a variety of cell
types, including liver, skin fibroblast, endothelia, epithelia,
Salmonella bacteria, and alveolar macrophages.[66–74] The
toxicity of nano-TiO2 to fish cells in vitro and to algae due
to ROS has also been suggested.[75,76]
Tests were conducted to measure ROS using a fluores-
cent probe by exposing Degussa P25 nano-TiO2 to brain
microglia (BV2).[63] Figure 7 shows measured concentra-
tions of O−·2 and H2 O2 as generated by the oxidative burst.
Significant release of O−· 2 did not happen until 60 min-
utes post-exposure (Fig. 7A). Comparatively, formation of
H2 O2 was rapid (Fig. 7B). Overall, the results of this study
suggest cellular and morphological expressions of free radi-
cal formation in response of brain microglia (BV2) to nano-
TiO2 . Fig. 7. (A) Increases in O−
2 were measured by the fluorescent probe
The toxic effect of TiO2 nanoparticles in mouse fi- MitoSOX Red. Cells, incubated in 2 µM MitoSOX (10 min, 37◦ C)
broblast cells (L929) has also been studied using homo- showed significant (p < 0.05) increases in fluorescence (measured
geneous and weakly aggregated TiO2 nanoparticles in at 510/580 nm) after 60 min exposure to g20 ppm P25, and flu-
orescence continued to increase for 150 min postexposure. (B)
aqueous solution.[77] As the concentration increased from
Extracellular release of H2 O2 was measured using OxyBURST.
3–600 µg mL−1 of TiO2 , the cellular shape became more Cells were incubated (30 min, 37◦ C) in 10 µg/mL OxyBURST in
spherical and levels of ROS and lactate dehydrogenase reduced-serum media, washed, and then exposed to P25 (2.5-120
(LDH) increased. The levels of methyl tetrazolium cyto- ppm). Significant (p < 0.05) increases of fluorescence (measured
toxicity (MTT), glutathione (GSH), and superoxide dis- at 508/528 nm) occurred in cells exposed to ≥ 10 ppm at 5 min
mutase (SOD) decrease; suggesting involvement of ROS in and continued to increase throughout the 120 min recording pe-
the cytoxicity of the TiO2 nanoparticles to L929 cells.[77] riod. (Reproduced from [63] with the permission of the American
Figure 8 shows TEM images of the internalization of Chemical Society)
TiO2 nanoparticels in L929 cells. Some particles were en-
closed in lysosomes while others were engulfed. This sug-
gests that incorporation of TiO2 nanoparticles into cellular in growth occurred at 1 g TiO2 L−1 . Nano-TiO2 suspension
membranes[78] caused an increase in the number of lyso- at 20 g L−1 were not acutely toxic to V. fischeri.[80]
somes and damaged some cytoplasmic organelles (Fig. 8).
Algae
A test of nano-TiO2 on green alga Desmodesmus subspi-
Ecotoxicologic studies
catus has been conducted using 25 nm and 100 nm TiO2
particles.[81] It was found that the size and crystalline form
Bacteria
of nano-TiO2 determined the toxicity on D. subspicatus.
Toxicity of nano-TiO2 has been tested on bacteria Vibria fis- Smaller particles were found to be more toxic to D. subspi-
cheri, Escherichia coli, and Bacillus subtilis.[79,80] High con- catus (72 hr EC50 = 44 mg L−1 ) than larger particles (72
centrations of nano-TiO2 are required to inhibit the growth hr EC50 >50 mg L−1 ). Recently, toxicity tests of nano-TiO2
of bacteria. For example, 72% reduction in the growth of E. (25–70 nm) and bulk-TiO2 were performed on Pseudokirch-
coli occurred at 5 g TiO2 L−1 . Comparatively, B. subtilis hap- neriella subcapitata.[77] Nano-TiO2 were more toxic than
pened to be slightly more sensitive and a similar reduction bulk-TiO2 . The EC50 values determined were 5.83 mg Ti
TiO2 nanoparticles in aquatic environment 1491

Fig. 8. TEM image of TiO2 nanoparticles internalization in L929 cells. (a) Normal cellular and mitochondrial morphology of cells
cultured in DMEM (control). (b) Increase in the number of lysosomes and disappearance of cytoplasmic organelles in L929 cells
cultured in media containing TiO2 nanoparticles (300 µg/mL) for 48 h. (c and d) Higher magnification of the agglomerated region
shown in b. (c) Lysosomes engulfed TiO2 nanoparticles in L929 cells by phagocytosis. Internalized TiO2 nanoparticles were present
primarily along and inside the ysosomes. (d) Apparent discontinuities in the cell membrane and cytoplasm were observed in L929
cells. (Reproduced from [77] with the permission of the American Chemical Society)

L−1 and 35.9 mg Ti L−1 for nano-TiO2 and bulk-TiO2 , re-
spectively. Interestingly, both nano- and bulk- TiO2 formed
aggregates during incubation (Figs. 9A and D). Phase con-
trast microscopy and red fluorescence confirmed the visibil-
ity of algal cells (Figs. 9B, C, E and F). Significantly, large
aggregates were observed that entrapped almost all algal
cells in the case of nano-TiO2 (Figs. 9 E and F). However,
in using culture with bulk-TiO2 , free algal cells in addi-
tion to cells trapped on small TiO2 aggregates were seen. It
seems aggregation of nano-TiO2 reduced the availability of
light to entrapped algal cells, thus inhibiting their growth.
It has been shown independently that adsorption of P. sub-
capitata onto surfaces of nano-TiO2 carried 2.3 times their
own weight in TiO2 particles.[82] Furthermore, kinetics and
extent of adsorption depended on the pH with maximum
adsorption occurring at pH 5.5.

Fig. 9. Aggregates of bulk and nano TiO2 particles in Pseudokirchneriella subcpitata culture. Aggregates of bulk TiO2 (A,B,C) and
nano (D,E,F) in test medium, as visible to the naked eye (A,D), in phase contrast microscopy (B,E) and fluorescence microscopy
*C,F). Arrows indicate algal cells. (Reproduced from [76] with the permission of Elsevier, Inc.)
1492 Sharma
Table 2. Body length, number of eggs inside the worm body, and number of offspring per individual of C. elegans after
exposure to different concentrations of TiO2 nanoparticles and bulk TiO2 . E. coli was added as a food to C. elegans. Each
test was repeated for four times and n = 425. All values were expressed as mean ± standard deviation (SD) of four replicates.
(Reproduced from [52] with the permission of Elsevier, Inc.)

Treatment Body length (µm) Eggs inside body Offspring per worm

Control 1126 ± 3 11.1 ± 1.2 28.4 ± 3.5

24.0 mg L−1 nano-TiO2 1080 ± 24 9.8 ± 2.3 21.7 ± 5.8
47.9 mg L−1 nano-TiO2 801 ± 82∗ 4.8 ± 1.9∗ 3.4 ± 3.1∗∗
95.9 mg L−1 nano-TiO2 760 ± 29∗∗ 2.1 ± 0.8∗∗ 0.9 ± 1.2∗∗
167.8 mg L−1 nano-TiO2 740 ± 15∗∗ 0∗∗ 0∗∗
167.8 mg L−1 nano-TiO2 (supernatant) 980 ± 72 8.9 ± 1.6 18.5 ± 2.4
24.0 mg L−1 bulk TiO2 975 ± 66 9.1 ± 3.0 22.5 ± 4.2
47.9 mg L−1 bulk TiO2 923 ± 10 7.9 ± 2.0 16.3 ± 5.5
95.9 mg L−1 bulk TiO2 812 ± 59∗ 5.8 ± 1.5∗ 5.6 ± 3.2∗∗
167.8 mg L−1 bulk TiO2 770 ± 48∗∗ 2.2 ± 1.3∗∗ 2.3 ± 2.6∗∗
∗
Significantly different from the control within each column at P < 0.05 level.
∗∗
Significantly different from the control within each column at P < 0.01 level.

Invertebrates Rainbow trout

Nano-TiO2 were exposed to terrestrial arthropods (Por-
cellio scaber, Isopoda, Crustacea) and feeding parameters,
weight change, mortality, and the activities of catalase and
glutathione-S-transferase were evaluated after 3 or 14 days
of dietary exposure.[83] It was found that the effects of nano-
TiO2 depended on total consumed quantity, size, and pre-
treatment of particles as well as on exposure concentration
and duration.
Daphnia magna as a test organism for aquatic inverte-
brates was exposed to nano-TiO2 .[84] A low mg L−1 range
of nanoparticles caused low toxicity, however the use of
high µg L−1 ranges gave some indication of chronic toxicity
and behavioral changes. Another study of toxicity of nano-
TiO2 was also conducted on Daphnia magna and Thamno-
cephalus platyurus.[80] The tests showed 60% mortality of
D. magna at 20 g L−1 TiO2 , but no toxicity forT. platyurus
was observed even at such a high TiO2 level. However il-
lumination of nano-TiO2 resulted in acute toxic effects on
daphnids.[81]
Nematode
Toxicity of nano-TiO2 and bulk-TiO2 were examined for
the nematode Caenorhabditis elegans with Escherchia coli as
a food source.[85] Nano-TiO2 and bulk-TiO2 has diameters
of 50 nm and 285 nm, respectively. The results are presented
in Table 2. No demonstrated toxicity of bulk-TiO2 to the
nematode was observed until the concentration was>95.9
mg L−1 . Comparatively, nano-TiO2 showed toxicity effects
at 47.9 mg L−1 . Significantly, the toxicity of nano-TiO2 su-
pernatant at 167.7 mg L−1 was not too different from the
respective controls. Furthermore, exposure of 47.9, 95.9,
and 167.8 mg L−1 nano-TiO2 gave lower numbers of off-
springs per worm than those exposed to bulk-TiO2 .
An in vitro study has been carried out by exposing rain-
bow trout (Oncorhynchus mykiss) to 0.1, 0.5, and 1.0 mg
L−1 nano-TiO2 for up to 14 days.[86] Exposure caused some
gill pathologies including odema and thickening of lamel-
lae. Levels of metals (Na+ , K+ , Ca2+ , and Mn2+ ) in tissues
did not change, but concentration-dependent changes of
Cu and Zn levels were observed, particularly in the brain.
Conclusions
Properties of nano-TiO2 differ from those of bulk-TiO2
of the same composition, which results in many novel ap-
plications of the nanoparticles. However, possible harmful
interactions of nano-TiO2 with biological systems cause
potential size-dependent toxic effects. The extent of ag-
gregation of nano-TiO2 in the aquatic environment deter-
mines the size of nano-TiO2 , which indicates whether nano-
TiO2 remains dispersed or produces larger-sized aggregates.
Though progress has been made in understanding the in-
fluence of pH, ionic composition, and ionic strength of
aqueous suspensions on aggregation, little is known about
the effects of the natural organic matter on the aggregation
of nano-TiO2 .
Furthermore, the effects of structure and properties of
humic substances in terms of aromaticity, conformation,
and polarity of aggregation of TiO2 needs examination.
Available data on the toxicity of nano-TiO2 in the aquatic
environment is limited. Future efforts should be made in
towards understanding the role of properties such as size,
shape, degree of agglomeration, chemical and catalytic of
particles contribute to toxic effects on bacteria, algae, inver-
tebrates, and vertebrate species. There is a possibility that
other toxicants can be associated with TiO2 nanoparticles
TiO2 nanoparticles in aquatic environment
and therefore likely synergistic effects must be examined for
additional toxicological concerns.
Acknowledgment
Author wishes to thank Drs. Mary Sohn and Ria Yngard
for useful comments on this paper. This paper has also been
greatly improved by the comments of the reviewer.
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