Iran Polym J
DOI 10.1007/s13726-014-0258-3
ORIGINAL PAPER
Fabrication and characterization of chitosan/poly(vinyl alcohol)
electrospun nanofibrous membranes containing silver
nanoparticles for antibacterial water filtration
Siavash Adibzadeh Saeed Bazgir
Ali A. Katbab
Received: 8 December 2013 / Accepted: 31 May 2014
Iran Polymer and Petrochemical Institute 2014
Abstract Electrospun nanofibrous membranes (ENMs)
were fabricated based on chitosan/poly(vinyl alcohol) (CS/
PVA) with a 70/30 mass ratio containing silver nanopar-
ticles (AgNPs) via the electrospinning method. AgNPs
were produced on the surface of CS/PVA nanofibers by
adding AgNO3 to a CS/PVA blend solution as a silver
rendering component. The presence of AgNPs in the
polymer blend solution was detected by UV spectropho-
tometry. The morphology of nanofibers before and after
cross-linking with glutaraldehyde was investigated by the
field emission scanning electron microscopy. The forma-
tion and size distribution of AgNPs onto the surface of
nanofibers were observed by transmission electron
microscopy and confirmed by energy dispersing X-ray
spectroscopy. As-spun and cross-linked CS/PVA nanofi-
bers revealed a smooth surface with diameters ranging
from 58 to 73 nm and 95 to 109 nm, respectively. The
effect of AgNP formation on the chemical structure of
nanofibers was explored by Fourier transform infrared
spectroscopy. Static and dynamic antibacterial filtration
efficiencies of CS/PVA ENMs, containing differing
amounts of AgNO3, have been tested against Escherichia
coli, a gram negative bacterium. The antibacterial assess-
ment results exhibited a significant increase in both static
and dynamic antibacterial filtration efficiencies of the
prepared CS/PVA ENMs by addition of AgNO3 as a bac-
tericidal agent.
S. Adibzadeh  S. Bazgir (&)  A. A. Katbab
Department of Polymer Engineering, Science and Research
Branch, Islamic Azad University, PO Box: 14155-4933,
Tehran, Iran
e-mail: bazgir@srbiau.ac.ir
Keywords Chitosan  Poly(vinyl alcohol)  Electrospun
nanofibrous membrane  Silver nanoparticle  Antibacterial
water filtration
Introduction
Conventional water purification and disinfection methods,
like chemical treatment and filtration by conventional
membranes, are not without problems. Chemical treatment,
defined as the addition of biocides like chlorine-based
components, leads to the production of carcinogen disin-
fection by-products (DBPs) [1]. In contrast, filtration of
microorganisms by conventional membranes leads to the
formation of biofilms, which significantly decrease the
quality of filtered water [2]. These problems in conven-
tional water purification and filtration systems make the
improvement of water filtration systems an attractive
research trend. Electrospinning has been known as a simple
and effective method in producing continuous non-woven
fibers with nanometric diameters [3, 4]. Electrospun
nanofibers, as a result of unique characteristics like high
surface area to volume, high aspect ratio, random orienta-
tion, high air and water permeability; have been widely
studied for different applications including tissue engi-
neering, drug delivery, and filtration [57]. As recently,
antibacterial water filtration using functionalized ENMs
has been investigateddue to high porosity, low pore size,
and high water fluxas an alternative to conventional
water purification and disinfection methods. Tan and
Obendorf [8] used nylon 6 ENMs to stabilize different N-
halamines as bactericides to improve the antibacterial
activity of the prepared ENMs. The obtained ENMs com-
posite showed high antibacterial activity and low N-hal-
amine leaching. Daels et al. [9] worked on the application
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of different biocides, like polychloride [(dimethylimino)(2-
hydroxy-1,3-propanediyl)] (WSCP) and AgNPs, have been
tested to bond antibacterial properties to polyamide
nanofibers for the removal of gram negative and gram
positive bacteria. WSCPs containing polyamide nanofibers
illustrate higher microorganism removal efficiency from
water samples between different bactericides. In other
works, CS-based ENMs were tried due to intrinsic anti-
bacterial properties and biocompatibility, as next genera-
tion filters for antibacterial water filtration applications [10,
11].
CS with a chemical structure of 1-4-linked-2-amino-2-
deoxy-b-D-glucopyranose is a biodegradable and biocom-
patible polycation acquired from chitin, the second most
abundant polysaccharide after cellulose [12]. CS, because
of the presence of amine groups in its backbones, shows
inherent antibacterial activity, as well as metal binding
properties in aqueous beds [13, 14]. Direct electrospinning
of CS is a challenging issue [15]. Internal hydrogen
bonding between hydroxyl and amine groups in CS back-
bones is one of the main reasons for the electrospinning
weakness of CS. The application of guest polymer with
good electrospinning ability, like PVA and PEO, is one
way to moderate the internal interaction between CS
backbones [1618]. In addition, solution properties such as
viscosity, ionic conductivity, surface tension, solvent type,
blend weight ratio, and polymer concentration are some of
the important factors which have direct effects on the
spinnability of the CS solution [19]. Another way to
improve the spinnability of CS is to lower CS solution
surface tension by using concentrated acetic acid as a
solvent. It is stated that, surface tension determines lower
and upper boundaries of the electrospinning window if
other factors are held constant. It is demonstrated that
although increasing the CS solution acidity leads to a slight
increase in solution viscosity, the CS solution surface
tension is decreased significantly [20, 21]. The CS can act
as a reducing agent to produce AgNPs from AgNO3. An
et al. [22] reported that the addition of AgNO3 to a CS/PEO
solution, as a silver rendering component, could produce
AgNPs containing CS/PEO nanofibers after electrospin-
ning, yielding higher antibacterial activity in comparison to
plain CS/PEO nanofibers. Moreover, toxicity of the silver
has been admitted to be significantly lower to human cells
than bacteria [23]. The Ag ion could interact with lone pair
electrons of hydroxyl groups in PVA backbones, as well as
amine and hydroxyl groups in CS. These interactions alter
Ag? to Ag, stabilizing Ag in the CS and PVA backbones
[24].
In the present study, CS/PVA ENMs, with and without
AgNPs, were fabricated onto a polyester substrate by
electrospinning technique. Ionic conductivity of CS/PVA
solutions was measured with different CS contents, as well
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as CS/PVA solutions with 70/30 mass ratios, including
differing amounts of AgNO3. The prepared CS/PVA ENMs
were then cross-linked using GA vapor as a cross-linking
agent. The effect of the addition of AgNO3 on the chemical
structure and morphology of the prepared nanofibers were
observed using the Fourier transform infrared spectroscopy
(FTIR) and field emission scanning electron microscopy
(FESEM) analyses, respectively. The formation of AgNPs
in the polymer solution and onto the surface of CS/PVA
nanofibers was illustrated by UV spectrophotometry and
transmission electron microscopy (TEM), respectively. The
presence of AgNPs on the surface of CS/PVA nanofibers
was confirmed by energy dispersing X-ray spectroscopy
(EDX). The leaching behavior of AgNPs from CS/PVA
ENMs was determined using both static and dynamic
methods. Static and dynamic antibacterial filtration effi-
ciencies of CS/PVA ENMs, with and without AgNPs,
against water samples containing E. coli, a gram negative
bacterium, have been evaluated. To our knowledge, this is
the first study which investigates different aspects of
chitosan-based electrospun nanofibrous membranes con-
taining silver nanoparticles capability to be utilized in
antibacterial water filtration.
Experimental
Materials
Medium molecular weight CS with a 7585 % degree of
deacetylation (DDA) was purchased from Sigma Aldrich.
PVA (72,000 molecular weight), acetic acid (99.5 % gla-
cial), GA (25 % in water), and silver nitrate powder were
provided by Merck. Distilled water was used. All chemi-
cals were used as received without any further purification.
Solution preparation and electrospinning
CS solution was prepared by dissolving CS powder in
90 % acetic acid/water solvent under extensive stirring
conditions at 80 C for 4 h. PVA powder was dissolved in
double-distilled water under gentle stirring conditions at
80 C for 4 h in order to produce a homogeneous solution.
AgNO3 was added to the CS/PVA solution with a 70/30
mass ratio to prepare blend solutions containing 0, 2, and 4
wt% AgNO3 and a polymer concentration of 3 wt%. The
obtained solutions were stirred for 5 h at 70 C to obtain a
homogeneous solution. The blend solutions were inserted
into a 60 mL syringe with a metallic needle (inner diameter
gage of 18) that connected to a high voltage power supply.
The voltage was adjusted to 18 kV and a solution flow rate
was set to 0.1 mL/h using a syringe pump (AMPALL,
Korea). A rotational steel collector which was covered by
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polyester substrate collected nanofibers onto the surface of
the substrate during electrospinning. Needle tip to collector
distance was adjusted at 15 cm.
Measurement and characterization
The ionic conductivity of the solutions was measured
using a CRISON conductimeter (GLP32, Spain). The
morphology of CS/PVA ENMs comprising of various
amounts of AgNO3 was observed using a Hitachi FESEM
(S-4160, Japan). The presence of AgNPs in the polymer
solution was shown using a UV spectrophotometer
(HACH-DR 5000, USA) that detected UV spectra with
waves ranging from 350 to 600 nm. The formation of
AgNPs onto the surface of the CS/PVA nanofibers was
observed by TEM (Zeiss-EM10C, Germany) and con-
firmed using EDX analyses. The average fiber diameter
and fiber size distribution for all ENMs were measured by
averaging the diameter of 50 random fibers using Image J
software. The mean pore size diameter of CS/PVA
membranes has been determined using image analysis
method [25]. CS/PVA ENMs were cross-linked in GA
vapor at room temperature for 24 h and heated for 18 h at
110 C to complete cross-linking and eliminate residual
GA. FTIR spectra of bulk CS and CS/PVA with a mass
ratio of 70/30, with and without AgNPs, were recorded
using a Thermo-Nicolet Nexus spectrometer (670 FTIR
Spectrometer, USA).
Leaching properties
The leaching properties of the CS/PVA nanofibers con-
taining AgNPs were measured using both static and
dynamic methods. Static silver leaching properties of CS/
PVA nanofibers containing 4 wt% AgNO3 were deter-
mined by immersing 10 mg of nanofibers containing Ag-
NPs into the phosphate buffer saline (PBS) and to
determine the amounts of silver trace the PBS sample
contained at different exposure times. In the dynamic
method, silver leaching from the CS/PVA ENMs contain-
ing 4 wt% AgNO3 was determined by filtering 100 mL of
PBS through the CS/PVA ENMs and consecutively mea-
suring the silver content released into the PBS after each
filtration. Four batches of 100 mL PBS were consecutively
filtered through the CS/PVA ENMs and the silver content
released was measured by the flame atomic absorption
spectrophotometry method (Varian Spectra AA-200). The
test was repeated three times.
Antibacterial assessments
The antibacterial properties of CS/PVA ENMs, with and
without AgNPs, were evaluated against PBS samples
Fig. 1 Ionic conductivity of the CS/PVA solution with different CS
contents and different AgNO3 contents
containing E. coli, a gram negative bacterium, using static
and dynamic antibacterial filtration methods. For the static
antibacterial test, 10 mg of nanofibers containing various
AgNO3 contents were added into tubes containing 10 mL
of PBS by pH 7.4 and inoculated with E. coli to prepare a
bacterial suspension with a concentration of 1.5 9 105
colony forming unit (CFU)/mL. A suspension of 0.1 mL
was removed and quickly poured into plates containing the
nutrient agar at different immersion times. The prepared
plates were incubated at 37 C for 24 h and the colonies
were determined using the pour plate method. The reduc-
tion in bacterial colonies was measured using the following
equation:
log reduction Log initial bacteria concentration
 Log final bacteria centration:
For the dynamic antibacterial test, 100 mL of bacterial
suspension with a 1.5 9 105 CFU/mL concentration was
prepared using the PBS as described before, and filtered
through the CS/PVA ENMs (diameters of 4.7 cm2) using a
dead end filtration unit. The filtration unit was autoclaved
at 121 C for about 15 min. The reduction in bacterial
concentration after each filtration was measured using the
pour plate procedure.
Results and discussion
Morphology of CS/PVA ENMs
The CS/PVA nanofibers with 70/30 mass ratios containing
AgNPs were fabricated onto the polyester substrate to
improve the mechanical characteristics of the ENMs nee-
ded for the water filtration application. The effect of
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Fig. 2 FESEM images of CS/
PVA nanofibers containing a 0,
b 2, and c 4 wt% AgNO3
varying CS contents in the CS/PVA solution on the ionic
conductivity of the blend solution, as well as the effect of
adding different amounts of AgNO3 into the CS/PVA
solution with a 70/30 mass ratio on the ionic conductivity
of the CS/PVA solution is depicted in Fig. 1. As shown in
Fig. 1 by increasing the CS/PVA ratio of the blend solu-
tion, the ionic conductivity is decreased. It is reported that
the viscosity of the CS/PVA solution is lowered by
increasing the CS content [26]. In addition, the addition of
AgNO3 into the blend solution leads to higher ionic con-
ductivity of the solution.
The low ionic conductivity of the CS/PVA solution with
a 90/10 mass ratio led to the formation of nanofibers with
beaded structures (results not shown). By decreasing the
CS/PVA mass ratio from 90/10 to 70/30, the ionic con-
ductivity of the CS/PVA solution was increased from
860 8 to 964 10 ls/cm which facilitated the forma-
tion of jet polymer during the electrospinning process and
subsequently contributed in the formation of bead-free
fibers. The FESEM images of CS/PVA nanofibers with
70/30 mass ratios containing 0, 2, and 4 wt% AgNO3 are
shown in Fig. 2. It can be seen from Fig. 2 that as-spun
nanofibers containing 0, 2, and 4 wt% AgNO3 show bead-
free structures with mean diameters of 73 15, 65 12
and 59 10 nm, respectively. A slight decrease in the
mean fiber diameters of CS/PVA nanofibers containing
AgNPs can be correlated with the higher ionic conductivity
of the solution, derived from the presence of AgNO3 in
polymer solution, as presented in Fig. 1. Also, the addition
of AgNO3 increases the viscosity of CS/PVA solution,
which could prevent fracturing of polymer jets that may
contribute in the formation of smooth and continuous
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nanofibers under synergistic effect of viscosity and ionic
conductivity [27].
In Fig. 3, the FESEM images and fiber size distributions
of CS/PVA nanofibers with 70/30 mass ratios containing
4 wt% AgNO3 before cross-linking (Fig. 3a, b) and after
cross-linking (Fig. 3c, d) are shown. It can be determined
from Fig. 3 that the mean fiber diameter is shown to
increase from 59 10 to 96 12 nm after cross-linking.
It is well-known that increases in mean fiber diameter,
along with white to yellow color changing, are character-
istic of a cross-linking reaction between hydroxyl and
amine groups of CS with aldehyde groups of GA [28].
Characterization of Ag nanoparticles
As functionalizing agent, the formation of AgNPs into the
blend solution and onto the surface of the CS/PVA
nanofibers was confirmed by UV spectra of the CS/PVA
solutions containing different amounts of AgNO3 and by
using EDX analyses, respectively (Fig. 4a, b). It can be
observed from Fig. 4a that by adding AgNO3 into the blend
solution, a broad peak appears in its UV spectra around
430 nm. The appearance of this peak is evidence of the
formation of AgNPs in the blend solution [29]. By
increasing the AgNO3 content from 0 (Fig. 4c) to 2
(Fig. 4d) and to 4 wt% (Fig. 4e), the peak intensity is
increased. This increase in AgNO3 causes an increase in
the AgNPs content of the solution. The EDX analysis was
used to confirm the presence of AgNPs on the surface of
nanofibers by detecting the appearance of a peak around
3 keV in the EDX graph, which is characteristic of the Ag
element (Fig. 4b) [30].
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Fig. 3 FESEM images of CS/PVA nanofibers containing 4 wt% AgNO3: a, b before cross-linking and c, d after cross-linking
In Fig. 5 the TEM images of CS/PVA nanofibers con-
taining 2 (Fig. 5a) and 4 wt% AgNO3 (Fig. 5b) are shown.
It can be seen from Fig. 5 that by increasing the AgNO3
content from 2 to 4 wt%, the mean diameter of the particles
increased from 10 4 to 34 10 nm without signifi-
cantly increasing the number of AgNPs. In Fig. 5b,
aggregation of the AgNPs into clusters on the fiber surface
can clearly be seen. It is found that AgNP formation starts
with the nucleation of reduced Ag? ions into Ag atoms
followed by the growth of Ag nuclei. The nucleation of the
AgNPs requires higher activation energy than the amount
needed for the growth stage of AgNPs [29]. It is reported
that increases of the CS content in CS/PVA/Ag? solution
up to 12.5 wt% lead to lowering number of stabilized
AgNPs and an increase in particle size [31]. In this study,
the CS content of the blend solution has been increased up
to 70 wt%, which provides higher amine and hydroxyl
groups in the polymer solution that could interact with the
Ag? ions by chelation and gathering together. These
attractions between Ag? and amine groups increase under
the electrostatic force during electrospinning. Moreover,
increasing the AgNO3 content in the solution provides
more Ag? ions that could be enclosed by chelation. Ngu-
yen et al. [31] have stated that large AgNPs are formed on
the electrospun nanofibers by diffusion and aggregation of
the residual amount of Ag ions after electrospinning during
the heat treatment. Furthermore, it is clearly shown that
AgNPs are formed on the surface of nanofibers. This can be
explained by the presence of higher amine groups on the
surface of nanofibers, where there is a higher CS content.
This is in line with the observations of Penchev et al. [32].
Phase separation of the CS and PVA in the CS/PVA
nanofibers is reported by Zhang et al. [19].
FTIR analyses
The FTIR spectra of CS bulk, as-spun CS/PVA nanofibers,
and CS/PVA nanofibers containing AgNPs are shown in
Fig. 6. From FTIR spectra of bulk CS it is evident that
absorption peaks at 1,599 and 1,661 cm-1 are attributed to
amide II and amide I, respectively. The CO stretching
band is shown at 1,025 cm-1 and the CH stretching band
is observed at 2,873 cm-1. An absorption peak at
3,376 cm-1 is characteristic of the OH and NH
stretching vibrations. FTIR spectra of CS/PVA nanofibers
show that the addition of PVA to CS in CS/PVA nanofibers
would sharpen the absorption peaks of CS/PVA nanofibers,
which allows for hydrogen bonding between OH groups
of PVA with NH and OH groups of CS backbones [19].
An additional absorption peak at 1,727 cm-1 is charac-
teristics of C=O bands with acetic acid solvent remaining
in the nanofibers. The comparison of FTIR spectra of as-
spun CS/PVA nanofibers, with and without AgNPs, dem-
onstrates that the addition of AgNPs significantly lowers
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the amine absorption peaks at 1,599 and 3,419 cm-1.
These changes in peak intensities of the amine groups
illustrate the consumption of NH2 groups and the
Fig. 4 a UV spectra of CS/PVA solution containing: c 0 wt%, d 2
wt%, e and 4 wt% AgNO3; and b EDX analyses of CS/PVA
nanofibers containing AgNPs
Fig. 5 TEM images of CS/PVA nanofibers containing: a 2 and b 4 wt% AgNO3
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stabilization of Ag by the formation of NAg groups
through the chelation process.
Leaching kinetics
The leaching behavior of CS/PVA nanofibers containing
4 wt% AgNO3 was evaluated using the static and dynamic
methods under equal condition. Figure 7 illustrates the
amounts of silver released into the PBS samples in both the
static and dynamic silver release tests. During the static
method, leaching kinetics of AgNPs containing nanofibers
were evaluated by immersing 10 mg of CS/PVA nanofibers
containing 4 wt% AgNO3 into 50 mL of PBS. The amount
of silver leached from the nanofibers at different exposure
times was determined by flame atomic absorption spec-
trophotometry. The static silver release test is demonstrated
in Fig. 7 and it is shown that the amount of silver released
is increased rapidly during the first 3 h of exposure time,
while its release trend is dropped for the 6 and 24 h
immersion times. The total Ag? content released into the
PBS from the CS/PVA nanofibers containing 4 wt%
AgNO3 during the 24 h time frame was just below 0.8 mg/L,
which is equal to 15 % of the total Ag content. The
obtained results are consistent with data reported by Zhao
et al. [27]. Cao et al. [33] observed that there was about
9 % of the silver release from sulfonated polyethersulfone
membranes, including AgNPs in de-ionized water over a
24 h time frame. The amount of silver release increased up
to 14 % in saline solution during the same time period. For
the dynamic leaching test, four consecutive batches of
100 mL PBS were filtered through the CS/PVA ENMs
containing 4 wt% AgNO3 and the silver content was
determined after each filtration. It was observed that the
concentration of the Ag ions lost during the filtration was
rather high at the beginning of the filtration, while this
amount dropped with further filtration, as shown in Fig. 7.
Indeed, about 10 % of the total Ag? was released from
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Fig. 6 FTIR spectra of CS
bulk, CS/PVA nanofibers, and
CS/PVA nanofibers containing
AgNPs
Fig. 7 Dynamic silver release test and static silver release test
ENMs containing 4 wt% AgNO3, with a mass of 35 mg,
during the 0.4 l filtration. Increasing of the Ag ions into the
PBS sample can be explained by the release of AgNPs,
which have lower stabilization degrees on the surface of
the nanofibers during the first and second filtrations. Zod-
row et al. [34] indicated that about 10 % of the total Ag
ions leached into the filtrate sample was originated from
immobilized AgNPs on the surface of the polysulfone
membrane. Although the silver release of the CS/PVA
ENMs decreased by further filtration, the percentage of Ag
released content was rather high which limited its appli-
cation for water filtration.
Antibacterial assessments
The antibacterial behavior of prepared CS/PVA ENMs was
explored using both static and dynamic filtration methods
as described earlier. In the static method, bactericidal
kinetics of CS/PVA nanofibers were measured by
Fig. 8 Log of E. coli concentration in PBS samples after soaking
10 mg of nanofibers containing a 0, b 2, and c 4 wt% AgNO3
immersing approximately 10 mg of CS/PVA nanofibers
with different AgNO3 contents into 10 mL of bacterial
suspension with a 1.5 9 105 CFU/mL concentration. The
results of the static antibacterial test for nanofibers con-
taining 0, 2, and 4 wt% AgNO3, are demonstrated in Fig. 8.
About 2 log reductions can be seen for plain CS/PVA
nanofibers. Desai et al. [10] reported almost the same log
reduction for CS/PEO nanofibers. The antibacterial activity
of cross-linked CS/PVA nanofibers is related to its Schiff
base imine functionality, which was produced after the
interaction between CS amine groups and GA carbonyl
groups during cross-linking [28, 35]. CS/PVA nanofibers
containing 2 wt% AgNO3 killed all bacteria within 60 min
exposure time. By increasing the AgNO3 concentration in
the blend solution from 2 to 4 wt%, time needed to com-
pletely eliminate all bacteria decreased from 60 to 30 min.
It is noticeable that static antibacterial efficiencies of the
CS/PVA ENMs containing 2 and 4 wt% AgNO3 can
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Fig. 9 Log reduction of E. coli concentration after dynamic filtration
of bacterial suspension through CS/PVA ENMs containing 0, 2, and 4
wt% AgNO3
partially be related to the silver release content during the
exposure time, according to the silver released content
during the 30 and 60 min exposure time, as illustrated in
Fig. 7 [28].
Dynamic antibacterial tests were performed using a dead
end filtration unit by passing 100 mL of PBS samples
containing 2 9 105 CFU/mL through CS/PVA ENMs
containing 0, 2, and 4 wt% AgNO3 after filtration once and
thrice, as indicated in Fig. 9. In this study, the plain CS/
PVA ENMs with a mean pore size diameter of 0.36 lm
showed less than a 0.5 log CFU/mL reduction after filtra-
tion once and thrice (Fig. 9), which was comparable with
the removal of less than 0.5 log CFU/mL after one pass
obtained with the CS/PEO nanofibrous membrane [10].
Mille et al. [36] indicated that a membrane with a pore size
of 0.2 lm had about 1.5 log CFU/100 mL reduction. Sadr
Ghayeni et al. [37] reported that by decreasing the mean
pore size of the membrane to 0.05 and 0.1 lm, the filtration
efficiency increased with 2 log CFU/100 mL. Daels et al.
[9] reported that a reduction of 2.2 log CFU/100 mL was
acquired with non-functionalized polyamide nanofibrous
membrane with pore sizes ranging from 0.2 to 0.4 lm after
hospital wastewater filtration. Although it has been repor-
ted that filtration of the solution containing micron to
submicron contaminants leads to the formation of a cake
layer on the surface of the filter consequently clogging the
filter which can increase filtration efficiencies of the
membranes, there has been a slight increase in the anti-
bacterial filtration efficiency of the plain CS/PVA mem-
brane after second and third filtration [38]. This can
possibly be attributed to the reduction in the volume of the
bacteria cells under pressure during filtration which is the
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main reason for separation of the bacteria with larger size
than membrane pore size through the filter [35]. Further-
more, it is shown that the presence of the CS and PVA has
positive effect on membranes resistance against fouling
[11, 39, 40]. Filtration results of ENMs containing 2 and 4
wt% AgNO3 illustrate about a 1.2 and 1.8 log CFU/mL
reduction after one pass (Fig. 9). It can be deduced that by
increasing the passing number from one to three, antibac-
terial filtration efficiencies of ENMs increase from 1.2 to
1.5 and from 1.8 to 2.3 log CFU/mL reductions for CS/
PVA ENMs containing 2 and 4 wt% AgNO3, respectively.
AgNPs also, have been found to be effective against bio-
fouling or adhesion of bacteria to the surface of the
membrane [37]. Daels et al. [9] have found that polyamide
nanofibrous membranes containing 1 and 3 wt% AgNPs
exhibited a bacteria reduction of about 3.2 and 4 log CFU/
100 mL, respectively. Also, it can be clearly seen that
antibacterial filtration efficiencies of ENMs per passing
number significantly decrease in both ENMs containing 2
and 4 wt% AgNO3 after the first filtration.
These reductions in antibacterial filtration efficiencies
could be due to the release of some stabilized AgNPs from
the surface of nanofibers into the bacterial suspension, as
well as the deactivation of some AgNPs by the collected
bacteria on the surface of the stabilized AgNPs on the
surface of CS/PVA ENMs during the first filtration. In
addition, it can be determined from the dynamic antibac-
terial filtration efficiencies of CS/PVA ENMs containing 0,
2, and 4 wt% that an increase in AgNO3 content from 0 to
2 wt% exhibits a higher increase in antibacterial filtration
efficiencies in comparison to the increase in AgNO3 con-
tent from 2 to 4 wt%. This could be due to the aggregation
of additional AgNPs produced by the increase in the
AgNO3 content. The aggregation of AgNPs onto the sur-
face of nanofibers leads to an increase in the size of AgNPs
and a decrease in the resulted AgNPs surface area to vol-
ume (Fig. 5b). Desai et al. [10] demonstrated that the
dynamic antibacterial filtration efficiencies of CS/PEO
ENMs do not show any dependency on the nanofiber
grammage, mean fiber diameter, or the CS DDA. While the
results of the dynamic antibacterial test imply that the
antibacterial filtration efficiencies of the CS/PVA ENMs
containing AgNPs have direct dependency on the content
and size of the stabilized AgNPs on the surface of the CS/
PVA nanofibers.
Conclusion
CS/PVA ENMs with 70/30 mass ratios containing different
amounts of AgNPs were successfully fabricated onto
polyester substrate as supported by the electrospinning
method. The CS/PVA ENMs with differing AgNO3
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contents were cross-linked using GA to improve mechan-
ical and chemical stability of nanofibers in aqueous beds. It
is observed that large AgNPs are formed mainly onto the
surface of CS/PVA ENMs. Static and dynamic silver
release contents of the prepared ENMs have been tested to
be about 15 and 10 % of the total Ag content, respectively.
The antibacterial properties of the CS/PVA nanofibers were
investigated against PBS samples containing E. coli using
static and dynamic antibacterial test methods. Results of
the antibacterial tests demonstrated that the addition of
AgNO3 significantly increased the antibacterial filtration
efficiencies of the CS/PVA nanofibers in both static and
dynamic antibacterial filtration methods. In the static
antibacterial test, time needed to completely remove all
bacteria significantly decreased by 30 min for CS/PVA
nanofibers containing 4 wt% AgNO3. Dynamic antibacte-
rial filtration efficiencies of CS/PVA ENMs increased from
below 0.5 to 1.2 and 1.8 log reductions after one pass with
the addition of 2 and 4 wt% AgNO3, respectively. The
results of this study thereby demonstrated the capability of
using CS/PVA ENMs containing AgNPs for antibacterial
water filtration and disinfection. Further study should be
conducted to control the silver release properties of ENMs,
which can limit the application according to the drinking
water standards of different countries, and to test the long
term performance of the ENMs containing AgNPs. In
addition, various functionalized ENMs with different bac-
tericides, including organic biocides, should be tested to
obtain a cost effective membrane with a proper balance
between antibacterial properties and leaching behavior.
Acknowledgments This work is the result of two years of research
in the Nanopolymer Laboratory of Islamic Azad University, Science
and Research Unit. The authors would like to thank Jessica Wilker-
son, Tooraj Adibzadeh, and Hamid Bornasi for their co-operations.
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