w a t e r r e s e a r c h 4 7 ( 2 0 1 3 ) 3 3 8 9 e3 3 9 8

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The influence of antiscalants on biofouling of RO membranes

in seawater desalination

Amer Sweity a, Yoram Oren a, Zeev Ronen b, Moshe Herzberg a,*

a
Department of Desalination and Water Treatment, Zuckerberg Institute for Water Research, Albert Katz International School for Desert
Studies, Ben Gurion University of the Negev, Sede-Boqer Campus 84990, Israel
b
Department of Environmental Hydrology and Microbiology, Zuckerberg Institute for Water Research, Albert Katz International School for
Desert Studies, Ben Gurion University of the Negev, Sede-Boqer Campus 84990, Israel

article info abstract

Article history: Antiscalants are surface active polyelectrolyte compounds commonly used in reverse
Received 4 January 2013 osmosis (RO) desalination processes to avoid membrane scaling. In spite of the significant
Received in revised form roles of antiscalants in preventing membrane scaling, they are prone to enhance biofilm
16 March 2013 growth on RO membranes by either altering membrane surface properties or by serving as
Accepted 19 March 2013 nutritional source for microorganisms. In this study, the contribution of antiscalants to
Available online 29 March 2013 membrane biofouling in seawater desalination was investigated. The effects of two
commonly used antiscalants, polyphosphonate- and polyacrylate-based, were tested. The
Keywords: effects of RO membrane (DOW-Filmtec SW30 HRLE-400) exposure to antiscalants on its
Reverse osmosis physico-chemical properties were studied, including the consequent effects on initial
Antiscalant deposition and growth of the sessile microorganisms on the RO membrane surface. The
Biofouling effects of antiscalants on membrane physico-chemical properties were investigated by
Fouling filtration of seawater supplemented with the antiscalants through flat-sheet RO membrane
Scaling and changes in surface zeta potential and hydrophobicity were delineated. Adsorption of
Biofilm antiscalants to polyamide surfaces simulating RO membranes polyamide layer and their
effects on the consequent bacterial adhesion was tested using a quartz crystal microbal-
ance with dissipation monitoring technology (QCM-D) and direct fluorescent microscopy.
A significant increase in biofilm formation rate on RO membranes surface was observed in
the presence of both types of antiscalants. Polyacrylate-based antiscalant was shown to
enhance initial cell attachment as observed with the QCM-D and a parallel plate flow cell,
due to rendering the polyamide surface more hydrophobic. Polyphosphonate-based anti-
scalants also increased biofilm formation rate, most likely by serving as an additional
source of phosphorous to the seawater microbial population. A thicker biofilm layer was
formed on the RO membrane when the polyacrylate-based antiscalant was used. Following
these results, a wise selection of antiscalants for scaling control should take into account
their contribution to membrane biofouling propensity.
2013 Elsevier Ltd. All rights reserved.

1. Introduction of 1000e3500 g/mol, typically consisting of one component or

a combination of polyphosphates, polyphosphonates, poly-
Antiscalants (AS) are scale inhibitors polyelectrolytes (Smith, acrylates, and dendrimeric polymers (Farahbakhsh et al.,
1967) with reported optimal molecular weights in the range 2004; Shih et al., 2006). AS have a superior effect avoiding

* Corresponding author. Tel.: 972 8 6563520; fax: 972 8 6563503.

E-mail address: herzberg@bgu.ac.il (M. Herzberg).
0043-1354/$ e see front matter 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.watres.2013.03.042
3390 w a t e r r e s e a r c h 4 7 ( 2 0 1 3 ) 3 3 8 9 e3 3 9 8
precipitation of scale forming salts even at low dosage by
preventing formation of crystal larger than a critical size
(preventing nucleation) as well as by modifying the surface of
larger crystals (Drak et al., 2000). In supersaturated salt solu-
tions supplemented with AS, a significant delay in the in-
duction time needed for precipitation is usually observed and
the quality of the feed water is not affected (Fritzmann et al.,
2007; Greenlee et al., 2009; Hasson et al., 1998, 2003; Gloede
and Melin, 2006; Darton, 2000). In general, AS play crucial
roles in maintaining efficient RO plant operations at the
highest possible recovery by preventing the need to replace
expensive membranes prematurely; eliminating or reducing
the use of hazardous acids; producing less concentrate,
reducing energy costs, as well as the downtime caused by
frequent membrane cleaning (Greenlee et al., 2009; Gloede
and Melin, 2006; Darton, 2000; Vrouwenvelder et al., 2000).
Despite the significant roles of AS, they are reported to
enhance membrane fouling, which is another setback for
the optimum operation of any RO desalination process
(Fritzmann et al., 2007; Greenlee et al., 2009; Vrouwenvelder
et al., 2000, 2008; Baker and Dudley, 1998; Subramani and
Hoek, 2008; Sadr Ghayeni et al., 1998; Kochkodan et al., 2008;
Abd El Aleem et al., 1998; Fletcher, 1994; Ouazzani and
Bentama, 2008; Chong et al., 2008). Accordingly, it is very
important to use the minimum possible dosage of AS in order
to avoid fouling enhancement at higher concentrations (Al-
Shammiri et al., 2000). AS were shown to induce biofilm for-
mation in RO systems by increasing the microbial growth
potential up to 10 times of their normal growth rate
(Vrouwenvelder et al., 2000, 2010).
Chemicals used in water pretreatment in RO operations can
promotes instability of colloids, particles and bacteria that can
increase fouling rate (Winters, 1997). Biofilm formation is a
progressive and developmental process initiated in bacterial
deposition and irreversible adhesion to surfaces, followed by
formation of micro-colonies encased in self-produced extra-
cellular microbial matrix (also termed as extracellular polymeric
substances e EPS), maturation, and finally, dispersion of the
bacterial cells back to their planktonic stage (OToole et al., 2000).
Deposition and attachment of the bacterial cells onto surfaces is
a critical step in the overall process of biofilm formation, which
is mediated by biological, physical, and chemical factors
(Donlan, 2002). These factors includes substratum properties
such as roughness, charge density and hydrophobicity (Diaz
et al., 2010; Park et al., 2005); a conditioning film of macro-
molecules on the surface; hydrodynamics forces (Eshed et al.,
2008; Purevdorj et al., 2002); solution chemistry such as ionic
strength, pH, and the presence of multivalent cations (Chen
et al., 2009; Chen and Walker, 2007; Rijnaarts et al., 1999); and
bacterial cell surface properties such as hydrophobicity,
expression of flagella and pili, lipopolysaccharides (LPS), and
EPS. The reversible attachment of bacteria involves weak forces
such as van der Waals, electrostatic and hydrophobic in-
teractions between the bacterial cell and the substratum
(Vanloosdrecht et al., 1987). Vrouwenvelder et al. showed how
AS can enhance biofouling in RO membrane applications
(Vrouwenvelder et al., 2000, 2010) by serving as phosphorous
and carbon source of nutrients under limited nutritional con-
ditions. While Vrouwenvelder et al. were focused on the nutri-
tional effects of different AS on biofouling phenomena, in this
work, a broad perspective including both biological and physico-
chemical effects of AS on biofouling of RO membranes were
investigated. One important AS effect includes the alteration of
physico-chemical properties of either the membrane or bacte-
rial surface due to AS adsorption, which consequently increases
attachment of bacterial cells to the membranes surface.
2. Materials and methods
2.1. Model bacterial strain and media
Vibrio fischeri, a well-known marine bacterium used in this
study, was obtained from Stritch School of Medicine in Loyola
University, Chicago (http://www.meddean.luc.edu/lumen/
deptwebs/microbio/kv/kvmain.php). This bacterium, a green
florescent protein ( gfp) chromosomally tagged mutant
KV2682, was grown on LB agar medium supplemented with
double the amount of NaCl (20 g/L), Trizma base (2-amino-2-
(hydroxymethyl)-1,3-propanediol) THAM 6.057 g/L, and
2.5 mg/L chloramphenicol. The strain was incubated and
grown at 25  C and 250 rpm to final OD600 nm of 1. Then, the
late exponential cells (after 5e6 h of incubation) were centri-
fuged (4  C, 2500 g, 20 min), washed three times, and re-
suspended to final OD600 nm of 0.1 with either filtered
seawater or with seawater supplemented with 20 mg/L of
each of the AS. Polyphosphonate- and polyacrylate-based AS
analysis is shown the supplementary material (Table S1).
2.2. Membrane preparation
A laboratory scale RO unit (Fig. 1) comprised a membrane
cross-flow cell, high-pressure pump, feed water reservoir,
chiller equipped with a temperature control system and PID
pH controller for dosing CO2 gas and a data acquisition sys-
tem. Permeate flow rate, conductivity, and pH were monitored
in each experiment. A high flux RO flat-sheet membrane SW30
HRLE 400 (Dow-Filmtec, USA) was compacted with deionized
water (DW) at a pressure of 60 bar before each experiment
until the permeate flux attained a constant value, after 24 h.
A pressure of 60 bar and a temperature of 25  C were kept
constant during all the experiments. After the compaction
stage completed, pretreated seawater sampled from Palma-
chim desalination plant (Palmachim, Israel) (after floccula-
tion, coagulation, and sand and micronic filtration), with or
without AS (20 mg/L) were filtered through the membrane for
24 h in the RO desalination lab unit. Then, the conditioned
membranes were kept in 4  C, in similar seawater used for
conditioning the membranes until further analysis of biofilm
formation or surface properties characterization has been
carried out. Usually, AS dosage in real RO installations varies
between 2 and 8 mg/L. The reason for choosing 20 mg/L can be
justified due to buildup of AS concentration at the tail end of
the RO cascade due to recovery.
2.3. Membrane surface properties
Prior to each biofilm formation experiment, after conditioning
the membranes with seawater in the presence or absence of
AS, membrane surface properties, i.e., surface zeta potential
 w a t e r r e s e a r c h 4 7 ( 2 0 1 3 ) 3 3 8 9 e3 3 9 8
Fig. 1 e Cross-flow, flat-sheet, RO desalination unit used for conditioning RO membranes with AS during desalination of
seawater.
and hydrophopicity, were measured. Membrane surface zeta
potential was measured using a streaming potential analyzer
(SurPass Elektrokinetic Analyzer) at 10 mM NaCl solution with
or without pretreatment with AS. For each solution, mea-
surements were done twice. During each measurement, each
run of the electrolyte solution flow proceeded in two di-
rections (right to left and then left to right). The zeta potential
of the RO membranes was calculated from the streaming
potentials using the HelmholtzeSmoluchowski equation with
the Fairbrother and Mastin substitution (Benavente and
Jonsson, 2000; Deshmukh and Childress, 2001). The hydro-
phobicity of the RO membrane was deduced from the contact
angle analysis method, whereby it is determined by the
captive bubble method (OCA, Data Physics). A droplet size of
air water with diameter of 0.4e0.5 mm was introduced to the
RO membrane surface after the antiscalant treatment stage in
the RO unit. Duplicated experiments, with five different
measurements of contact angle, were carried out for each of
the treated membranes to obtain at least 10 measurements of
contact angle for each set of conditions. In addition, X-ray
photoelectron spectroscopy (XPS) was used to verify the effect
of AS addition on the elemental composition of the polyamide
surface membrane and properties after treating the mem-
brane surface with seawater supplemented with or without
AS. Elemental composition XPS analysis was deduced from
deconvoluted spectra of carbon, nitrogen, oxygen, and phos-
phorous binding energies on membrane surface presented in
the Supplementary material. Figures S1eS4, for pristine
membrane as well as membranes treated with seawater, and
seawater supplemented with either polyacrylate- or poly-
phosphonate-based AS, respectively.
2.4. Bacterial physico-chemical properties
The electrophoretic mobility of the bacterial cells was
measured by zeta potential analyzer (ZetaPlus 1994, Broo-
khaven instruments Co., Holtsville, NY). Cells were washed in
3391
10 mM NaCl with and without antiscalant (20 mg/L). The
relative hydrophobicity of the cells was determined by the
adhesion of microbial cells to hydrocarbon droplets test
(MATH test) with n-dodecane (Pembrey et al., 1999).
2.5. Monitoring bacterial attachment with quartz
crystal microbalance with dissipation (QCM-D)
Polyamide sensors mimicking RO membrane surfaces were
used in E4 module QCM-D (Q-Sense, SWEDEN) for analyzing
the effect of AS on bacterial deposition and attachment. All
QCM-D experiments were performed at flow-through condi-
tions using a digital peristaltic pump (IsmaTec Peristaltic
Pump, IDEX) operating in a pushing mode. The flow rate of the
working solution in the QCM-D flow cell was 150 mL/min. The
following solutions were injected sequentially to the QCM-D
system: (i) double distilled water baseline for 20 min; (ii)
0.2 mm filtered seawater for 20 min; (iii) seawater supple-
mented with 20 mg/L AS; (iv) bacterial suspension in 0.2 mm
filtered seawater with or without AS for 20 min; and (v) finally,
these steps were repeated in a reverse order. When no AS was
supplemented, step (ii) was injected for 40 min. The adsorp-
tion kinetic curves were made by Q-Tools software (Q-SENSE,
Sweden). This software adjusts the incoming and outgoing
electrical currents, regulates the amplitude of the oscillation
and controls the temperature according to the temperature
set. The variations of frequency shift (Df, Hz) and dissipation
factor (DD) were measured for five overtones (n 3, 5, 7, 9 and
11) and the 7th overtone is presented.
2.6. Bacterial deposition experiments using parallel plate
flow cell
After treating both the RO membranes and the bacterial cells
with AS, three sets of duplicated deposition experiments were
carried out to characterize the effect of AS treatment on the
deposition and attachment of the bacterial cells on the RO
3392 w a t e r r e s e a r c h 4 7 ( 2 0 1 3 ) 3 3 8 9 e3 3 9 8
membrane surface. AS concentration used to treat bacterial
cells and RO membranes was 20 mg/L. In each set of experi-
ments, characterization of the initial bacterial deposition to
the RO membrane surface was done by placing a piece of each
of the treated RO membranes in a parallel flow cell (FC81,
Biosurface Technologies, Bozeman, Montana). Then, the
treated bacterial suspension was injected into the flow cell
(140.6 mm long  12.7 mm wide  0.20 mm deep) at a velocity
of 8.5 cm/s. The accumulated bacteria on the membrane
surface were monitored with a fluorescent microscope (Zeiss
AXIO Imager) with magnification of 10X and every image was
taken at fixed spot of the membrane. The monitored spots on
the membrane coupon were visualized every 20 min and ac-
quired with a CCD camera. At least 5 pictures were taken
every 20 min, and the fluorescent signals of the bacteria were
manually counted on a rectangular viewing area of
861  650 mm, which was recorded by the CCD camera at a
magnification of 10. Before snapping the pictures, the same
background solution as in the experiment (without bacteria)
was injected into the flow cell for 20 s for washing the sus-
pended bacteria in order to visualize only the attached cells.
This way, each set of experiment was carried out for three
hours and the gradual increase in the number of deposited
bacteria was recorded and the number of deposited bacteria
per cm2 of the observed RO membrane surface was calculated.
As mentioned, at each time point, 5 different spots on the
membrane were visualized and the number of the deposited
cells was averaged, calculated per unit of area and normalized
to the initial cell concentration in each experiment. The
deposition coefficient was calculated as the number of
deposited cells per min per cm2 and normalized to the injected
bacterial cell concentration (Vanoyan et al., 2010).
2.7. Biofilm growth experiments on RO membranes in a
parallel plate flow cell
The conditioned RO membranes from the RO lab desalination
unit (Fig. 1) were used for biofilm growth experiments, which
were conducted in a parallel plate flow cell (FC81, Biosurface
Technologies, Bozeman, Montana). Seawater with or without
AS (100 mg/L) were injected into the flow cell, which was
occupied with SW30 membrane. It should be mentioned that
enhanced biofilm growth experiments were conducted with a
relatively high concentration of AS in order to compare be-
tween the AS being used and to achieve biofilm growth within
a reasonable experimental period. The flow cell dimensions
were 140.6 mm long  12.7 mm wide  0.20 mm deep and a
flow velocity of 8.5 cm/s was kept constant for ten days period
of biofilm growth. The AS concentration (100 mg/L) in the
inoculum was relatively high, in order to boost the bacterial
growth during the relatively short experimental period. The
microbial inoculum used for this experiment, was bacterial
culture cultivated for four months prepared as following:
sterile flasks (250 mL) with 150 mL of Palmachim desalination
plant treated seawater supplemented with AS were incubated
for 4 months at 30  C and 250 rpm. A weekly replacement of
140 mL of seawater with or without antiscalant was carried
throughout the 4 months period in order to create a selective
pressure for the growth of the antiscalant-degrading bacteria.
Each of the cultures was used as inoculum for the biofilm
growth experiments according to the antiscalant being
analyzed. At the end of each experiment, the membrane was
collected for confocal laser scanning microscope (CLSM)
analysis. Prior to the CLSM, pieces of the membranes
(5 mm  5 mm) from the flow cell were cut from the middle of
the membrane coupon and fixed immediately for all runs,
without drying the membranes. The fixations of the fouled
membranes were done by adding 0.05 M sodium cacodylate
buffer supplemented with 2% glutaraldehyde for 1 h. Then the
fouled membranes were stained with concanavalin A (ConA)
conjugated to Alexa Fluor 633 (Invitrogen, Israel) and with
propidium iodide (PI) for probing EPS and cells, respectively.
Microscopic observation and image acquisition were per-
formed using Zeiss-Meta 510, a CLSM equipped with Zeiss dry
objective LCI Plan-NeoFluar (25X magnification and numerical
aperture of 0.8). CLSM images were generated using the Zeiss
LSM Image Browser. Gray scale images were analyzed, and the
specific biovolume (mm3/mm2) was determined with IMARIS
v7.5 software (IMARIS Bitplane, Zurich, Switzerland). Table 1
summarizes the types of bacterial cultures used in the
different deposition and biofouling experiments.
3. Results and discussion
3.1. Effect of antiscalants on membrane surface
properties
The effect of AS on the surface physico-chemical properties of
the Filmtec SW30 RO membranes was determined by filtration
of Palmachim desalination plant feed seawater with or
without AS through the membranes for 24 h (Fig. 1).
3.1.1. Zeta potential effect
For all cases, at 10 mM NaCl, membranes surface zeta po-
tential was negative for pH values above 3.8 (Fig. 2A). At lower
pH, both AS increased membrane positive charge: a neutral
zeta potential was observed for polyacrylate-based AS at pH
below w3.5 and slightly higher positive zeta potential was
observed for the membrane treated with polyphosphonate-
based AS. Clearly, when the membrane was not exposed to
any of the AS, the membrane was more negatively charged for
the entire pH range.
Table 1 e Types of bacterial cultures used in this research
study.
Experiment Type of culture
Bacterial physico-chemical V. fischeri pure culture
properties
Bacterial deposition and V. fischeri pure culture
attachment using QCM-D
Bacterial deposition and V. fischeri pure culture
attachment experiments
using parallel plate flow cell
Biofilm growth experiments Natural microbial
on RO membranes in a consortium isolated
parallel plate flow cell from a fouled RO
membrane
 w a t e r r e s e a r c h 4 7 ( 2 0 1 3 ) 3 3 8 9 e3 3 9 8
3.1.2. Hydrophopicity effect
Using captive bubble contact angle method (Zhang and
Hallstrom, 1990) enabled us to deduce the effect of
polyacrylate-based and polyphosphonate-based AS under
most realistic conditions: upon exposing to seawater in the
absence of AS, the membrane surface was relatively hydro-
philic (Fig. 2B) with a contact angle of 32  1.5 . Interestingly,
when exposed to seawater in the presence of AS, a significant
increase in hydrophobicity of the RO membrane surface was
observed with contact angle measurements of 54.3  5.4 , and
42  4.3 for the membranes treated with polyphosphonate-
based and polyacrylate-based AS, respectively (Fig. 2B).
While polyacrylates are linear with amphiphilic structure, the
positive shift in zeta potential and the increased hydropho-
bicity of the membrane exposed to polyacrylate-based AS,
implies that carboxylic moieties on this AS interact with the
RO membrane, probably chemically cross-bridged with diva-
lent cations as Ca2 with the hydrophobic tail of this AS
exposed to aqueous surrounding environment. It should be
mentioned that zeta potential analysis of the membranes was
conducted in 10 mM NaCl after treating the membrane with
seawater with or without AS. The results imply insignificant
possible washout of the AS from the membranes during the
streaming potential analysis. Since polyphosphonate and
aminophosphonates, which mainly comprise the poly-
phosphonate-based AS, are smaller in their size and hydro-
philic than polyacrylates, almost no effect on membrane
hydrophobicity was observed. However, interactions of both
of carboxylic and phosphate moieties of the polyphosphonate
and the membrane surface interaction will be facilitated by
Ca2 cations (Guo and Severtson, 2004; Petit-Agnely et al.,
2000; Skwarczynski et al., 2010; Tsiourvas et al., 1997).
3.1.3. XPS analysis
The SW30 HRLE 400 polyamide membranes treated with
seawater supplemented with AS were characterized using
XPS to monitor the presence of AS fingerprinting on the
membrane surface as shown in Table 2. XPS is a sensitive
surface technique providing analysis of surface chemical
groups in a depth range of about 5e10 nm (Ferjani et al., 2000;
Williard, 2007). From the deconvoluted binding energy
10
Blank
Zeta potential, mV
0 CA
PP
-10
-20
-30
-40
2 4 6 8
pH
Fig. 2 e The effect of antiscalant treatment (20mg/L) on membrane surface charge and hydrophobicity: polyphosphonate-
based (PP) and polyacrylate-based (CA) AS were tested. (A) Filmtech-SW30 RO membranes surface zeta potential plotted as a
function of the pH in a background solution of 10 mM NaCl; (B) Captive air bubble contact angle on the surface of Filmtech-
SW30 RO membrane.
3393
spectra, five different types of carbon bonds, two different
types of nitrogen bonds, three types of oxygen bonds, and two
types of phosphorous bonds were revealed. The different
peaks of the binding energies (BE), the related bonds found in
previous studies (Ferjani et al., 2000; Williard, 2007; Boussu
et al., 2007; Liu et al., 2006; Tang et al., 2007, 2009), and our
suggested origin of the peaks (membrane or both membrane
and AS) are summarized in Table 2. As it was impossible to
distinguish between bonds originated from AS and the
membrane due to similarity of the bonds being detected, ev-
idences for the AS adsorption to the membrane were absence
of the aromatic CeC/CeH peak of 285 eV and the CH2eNCO
peak of 288 eV, only for the membrane treated with poly-
phosphonate-based AS (Table 2). In addition, for the mem-
brane treated with carboxylic acid-based AS, the nitrogen
peak that is attributed to amine group (with binding energy BE
between 400.76 and 400.85) was not detectable, probably due
to interaction of carboxylic acids with a surface amine group.
For the oxygen peaks, as presence of the three main types of
oxygen was detected on all membranes, and therefore oxygen
bonds were not useful to determine the presence of adsorbed
antiscalant. Still, the absence of O 1s peak at BE around 529,
rules out the presence of any metal oxides on the membrane
surface. Regarding the phosphorous bonds, the virgin mem-
brane had the least amount of phosphorous: the atomic per-
centage of phosphorous on the membranes were 0.19%, 0.3%,
0.26% and 0.54% for the virgin membrane, membrane treated
with seawater, membrane treated with seawater and poly-
acrylate-based AS and membrane treated with seawater and
polyphosphonates-based AS, respectively (Table 2). As ex-
pected, the membrane treated with polyphosphonate-based
AS showed the highest percentage of phosphorous.
3.2. Effect of antiscalants on bacterial physico-chemical
properties
3.2.1. Hydrophopicity effect
The effect of AS on the relative hydrophobicity of the V. fischeri
KV2682 bacterial cells was analyzed. Interestingly, when
treated with 20 mg/L polyphosphonate-based AS (PP), these
bacteria showed lower hydrophobicity compared to the
Contact angle ,Degrees
60
40
20
10 Blank CA PP
Membranes condtioned
 3394
Table 2 e Elemental compositions were computed based on C (1s) N (1s), O (1s), and peaks, which are centered around 532, 399, and 284 eV, respectively. BE refers to the
peak binding energy and % At refers to the elemental percentage on the surface of the membrane for 4 different membranes. Samples are pristine membrane, membrane
treated with seawater only, membrane treated with seawater supplemented with carboxylic acid-based AS and a membrane treated with seawater supplemented with
polyphosphonate-based AS.
Suggested bond Pristine membrane Seawater w/o AS Seawater CA Seawater PP

Peak BE At %. Suggested origin Peak BE At %. Suggested Peak BE At %. Suggested origin Peak BE At %. Suggested origin

w a t e r r e s e a r c h 4 7 ( 2 0 1 3 ) 3 3 8 9 e3 3 9 8
origin

C1S scan
Aliphatic CeC/CeH 284.65 52.51 Membrane 284.71 49.39 Membrane 284.62 49.48 Membrane or AS 284.59 46.32 Membrane or AS
CeO/CeN 286.31 22.12 Membrane 286.45 25.55 Membrane 286.38 22.07 Membrane 286.13 39.82 Membrane
COO orCH2eNCO 288.14 10.07 Membrane 288.4 11.26 Membrane 288.43 11.78 Membrane or AS Non-detectable: covered by layer of AS
Aromatic CeC/ 285.41 10.39 Membrane 285.49 9.94 Membrane 285.43 11.57 Membrane Non-detectable: covered by layer of AS
CeH stretch
O]CeN 287.33 4.9 Membrane 287.6 3.86 Membrane 287.44 5.09 Membrane 287.86 13.86 Membrane (shifted)
N1S scan
O]CeN 399.79 67.08 Membrane 400.04 92.69 Membrane 400.04 100 399.74 65 Membrane
Amine 400.85 32.92 Membrane 401.49 7.31 Membrane Non-detectable: interactions of COO 400.76 35 Membrane
(shifted) or with
ammonium amine groups
O1S scan e no metal oxides (absence of peaks at BE 529)
COO or OH 532.59 42.3 Membrane 532.54 58.78 Membrane 532.38 41.5 Membrane or AS 532.47 62.51 Membrane or AS
H 2O 533.43 31.8 Moisture 533.69 22.67 Membrane 533.45 30.52 Moisture 533.76 17.33 Moisture
O]CeN 531.53 25.9 Membrane 531.41 18.55 Membrane 531.42 27.98 Membrane or AS 531.17 20.16 Membrane or AS
P2p scan
CeP or PO 133.57 0.19 Membrane 133.74 0.30 Membrane 133.62 0.26 Membrane and 132.74 0.54 Membrane and
and seawater seawater seawater AS
 w a t e r r e s e a r c h 4 7 ( 2 0 1 3 ) 3 3 8 9 e3 3 9 8
polyacrylate-based AS as well as to the case without AS
(Fig. 3A). The values for the percent of partitioning between
the aqueous and the organic phase were 17%  0.62, in the
absence of the AS; 14%  0.42 in the presence of
polyphosphonate-based AS; and 28%  0.54, in the presence of
polyacrylate-based AS (Fig. 3A).
3.2.2. Zeta potential effect
The electrophoretic mobility of the V. fischeri KV2682 cells
treated with different AS is shown in Fig. 3B. The results show
no significant effect on the zeta potential of this strain by the
presence of both AS, with zeta potential values for all cases in
the range between 35 and 38 mV. Similar to the membrane
surface, changes in bacterial cell hydrophobicity in the pres-
ence of polyacrylate- and polyphosphonate-based AS are also
affected by the chemical nature of these compounds and the
effects are similar. Since bacterial cells were washed with
10 mM NaCl and in the absence of Ca2, prior to their zeta
potential measurements with the different AS, no change in
zeta potential was observed, in contrast to the membrane
surface, significantly affected by prior conditioning filtration
step of seawater in the RO lab unit.
3.3. Antiscalants effects on bacterial deposition
In order to study bacterial deposition and the related physical
and chemical interactions, model bacterium with defined
properties must be used. Clearly, a pure strain does not
represent the actual behavior of natural microbial consortium
in the environment but it could be the guidance for such
phenomena. The selected model strain, V. fischeri KV2682, is a
representative marine bacterium and its expression of a green
florescent protein ( gfp) allows precise microscopic tracking.
Recently, Naidu et al. (2013) used V. fischeri to investigate the
role of microbial activity in biofilter used as a pretreatment
stage for seawater desalination.
Here, the effects of AS on bacterial attachment were con-
ducted on a polyamide coated sensor in a QCM-D flow cell.
First, a baseline is achieved with double distilled water (DDW)
for 20 min to assure that the polyamide coated sensors is
absolutely clean. Then, filtered seawaters were injected as a
background solution, followed by injection of seawater with or
without antiscalants used to condition the polyamide surface.
After conditioning the surface with seawater in the presence
or absence of antiscalant, bacterial suspension was injected
under similar aquatic conditions. Frequency shift of the
Fig. 3 e The effect of antiscalant treatment (20 mg/L) of polyphosphonate (PP) and polyacrylate (CA) based AS on the relative
hydrophobicity (A) and the zeta potential (B) of Vibrio fischeri KV2682 bacterial strain in 10 mM NaCl at ambient pH of 6.2.
3395
sensor is shown later in this study, to be positively related to
the amount of attached/detached bacteria to/from the sensor
as confirmed by direct fluorescent microscopy. The results in
Fig. 4 show the AS effects on the adherence of the bacterium V.
fischeri KV2682. In the presence of the polyacrylate-based AS,
the bacteria cells attached to the sensor at higher extent
compared to their attachment in the presence of the
polyphosphonate-based AS, since a higher decrease of the
frequency was observed for the case using polyacrylate-based
AS. These results can be related to the combination of higher
hydrophobicities of the bacterial cells and polyamide surface
treated with polyacrylate-based AS as well as to the more
positive zeta potential of the AS treated membrane (Figs. 2 and
3). Yet, we do not have an explanation for the lower adhesion
of bacteria after treatment with polyphosphonate-based AS,
where in fact, the bacteria were excluded from the poly-
phosphonate treated surface, while attracted to the poly-
acrylate treated one. Possible reason may be the lower
hydrophobicity of the bacteria treated with polyphosphonate-
based AS as shown in Fig. 3A. Careful interpretation of the
frequency shift acquired during bacterial deposition on QCM-
D sensors should be taken into account, as the frequency can
be influenced by cell surface morphology with some cases,
where frequency shift is not positively related to the increase
of bacterial adhesion (Marcus et al., 2012). Therefore a positive
relation between deposition of bacterial cells and frequency
shift was established: similar trend for the antiscalant effect
on the bacterial cells attached to the RO membrane surfaces
was observed using direct fluorescent microscopy with this
bacterium as shown in the inset graph of Fig. 4.
3.4. The effect of antiscalants on biofilm formation
Biofilm growth experiments conducted using parallel plate
flow cell were conducted and the effect of the polyacrylate-
and polyphosphonate-based AS on biofilm formation by nat-
ural microbial consortium was delineated. As already
mentioned, prior to the injection of the seawater, with or
without the AS, inoculums from the incubated seawater were
supplemented to the flow cell in order to boost the biofilm
growth process. CLSM analysis of membrane samples taken
from the parallel plate flow cell shows that addition of anti-
scalant promoted biofilm growth on the surface of the RO
membrane (Fig. 5). Specific biovolume analysis of the biofilms
using IMARIS software (Bitplane, Switzerland) on the mem-
branes showed that polyacrylate treated membrane had more
3396 w a t e r r e s e a r c h 4 7 ( 2 0 1 3 ) 3 3 8 9 e3 3 9 8

DDW SW SW+AS Bacteria SW+AS SW DDW

3.0x10
0 2.5x10

2.0x10

-6
-5

Frequency, Hz X10
1.5x10

1.0x10
-10
5.0x10
0.0
-15 SW CA
Treatment
PP

-20
PP
-25
CA

-30 Blank

0 20 40 60 80 100 120 140

Time, Minutes
Fig. 4 e The effect of antiscalant addition on the attachment of bacterial cells to polyamide coated QCM-D sensor as
measured by change in frequency of oscillation of the 7th overtone. Inset graph describes the deposition coefficient for GFP
tagged Vibrio fischeri on SW30 RO membrane surfaces during cross-flow of 2 mL/min in a parallel plate flow cell (FC81 flow
cell, Biosurface Technologies, Bozeman, Montana).

biomass in comparison to the case of polyphosphonate
treated membranes and the membrane without antiscalant
(Fig. 5). While both polyacrylate- and polyphosphonate-based
AS increased biofilm formation, the increased bacterial
attachment analyzed for polyacrylate-based AS induced more
effectively biofilm formation process. Polyphosphonate, on
the other hand, had an opposite effect on bacterial attach-
ment that could not explain the moderate elevation in biofilm
formation as presented for this case in Fig. 5. The moderate
increase in biofilm formation in the presence of poly-
phosphonates could be attributed to the supplement of a
limiting nutritional element for microbial growth under
phosphorous limiting growth conditions in seawater (unde-
tectable level of total P). Note that in real RO installations, a
lower concentration of AS will be used, but still may reach
close to the levels used in this study, due to seawater desali-
nation recovery of around 50%. Likely, the lower AS concen-
tration being used, but still efficient in avoiding membrane
scaling, the lower biofouling side effects will rise. In should be
mentioned that more study need to be done in order to un-
derstand the contribution of each antiscalant to biofilm
growth. This can be achieved by measuring continuously
other biofilm growth-related parameters, including the pres-
ence of adenosine triphosphate (ATP) in the biofilms, and

Fig. 5 e Left panel represents three-dimensional reconstructed images acquire from CLSM using Imaris Bitplane software
(the red colour represents biomass and the green colour represents EPS) of the fouled SW30 RO membrane surface after
injecting seawater without AS (A); seawater supplemented with polyacrylate AS (B); and seawater supplemented with
polyphosphonate AS (C). The resolution of the perspective images is 450 3 450 mm. Right panel (D) represents the amount of
biomass per unit area of attached bacterial cells and adsorbed EPS in the biofilm layers. (For interpretation of the references
to colour in this figure legend, the reader is referred to the web version of this article.)
 w a t e r r e s e a r c h 4 7 ( 2 0 1 3 ) 3 3 8 9 e3 3 9 8
phosphorous and total organic carbon (TOC) concentration in
the feed and effluent water of the parallel plate flow cell.
4. Concluding remarks
While scale formation in RO desalination systems can be suc-
cessfully prevented by AS, they can enhance biofilm formation
on RO membranes during seawater desalination in several
ways, at different stages of biofilm growth. Polyacrylate-based
AS were shown to enhance biofilm formation, most likely by
altering the physico-chemical properties of the RO membranes
such as hydrophobicity and surface charge, which in turn
promote the initial deposition and attachment of bacterial
cells. Polyphosphonate-based AS were shown to contribute
membrane biofouling probably by acting as a phosphorous
source of nutrients under the phosphorous limitation condi-
tions prevailing in seawater. Comparing these two effects
shows that the physico-chemical effects of AS can severely
increase RO membrane biofouling. Based on the findings of this
work, it is strongly recommended that AS should be screened
for their biofouling contribution and related enhancing steps
such as particulate and microbial attachment, as well as
nutrient source, prior to their application.
Acknowledgements
This research study was funded by the Israel Governmental
Authority for Water and Sewage and by Albert Katz Interna-
tional School for Desert Studies (AKIS) in Ben-Gurion Univer-
sity of the Negev, which partially supported Amer Sweitys
PhD fellowship.
Appendix A. Supplementary data
Supplementary data related to this article can be found at
http://dx.doi.org/10.1016/j.watres.2013.03.042.
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