Desalination 485 (2020) 114457

Contents lists available at ScienceDirect

Desalination
journal homepage: www.elsevier.com/locate/desal

Fouling process of membrane distillation for seawater desalination: An T

especial focus on the thermal-effect and concentrating-effect during
biofouling
Longjie Jiang, Lin Chen , Liang Zhu
⁎ ⁎

Key Laboratory of Integrated Regulation and Resources Development of Shallow Lakes, Hohai University, Nanjing 210098, China
College of Environment, Hohai University, Nanjing 210098, China

GRAPHICAL ABSTRACT

ARTICLE INFO ABSTRACT

Keywords: Membrane distillation (MD) is a promising seawater desalination technology for remote areas, although it is still
Membrane distillation limited by membrane fouling. In this study, a lab-scale direct contact membrane distillation (DCMD) system was
Seawater desalination designed for seawater desalination, operating in concentrating and non-concentrating modes to characterize the
Biofouling fouling process. Systematic analysis showed that scaling and biofouling are dominant fouling types in MD
High throughput sequencing
system, although the high temperature and salinity can inhibit the formation of biofouling layer. Biofouling can
Concentrating-effect
accelerate the formation of scaling, and the mixed foulants can block the membrane pores, leading to significant
flux drop. With the application of high throughput sequencing, Proteobacteria, Bacteroidetes, Firmicutes and
Planctomycetes were found to be the most abundant phyla. Microbial community succession was revealed during
biofilm formation, in which Proteobacteria, Planctomycetes and Bacteroidetes played important roles. Additionally,
a higher abundance of Firmicutes was observed in concentrating mode, representing the selective pressure from
the heating and concentrating process. Finally, a three-phase model was suggested to describe the fouling
process, which may reveal the vital fouling stage, and assist the development of anti-biofouling approaches in
MD operation.

⁎
Corresponding authors at: Key Laboratory of Integrated Regulation and Resources Development on Shallow Lakes, Ministry of Education, Hohai University,
Nanjing 210098, China.
E-mail addresses: chen_lin@hhu.edu.cn (L. Chen), hhuzhuliang@163.com (L. Zhu).

https://doi.org/10.1016/j.desal.2020.114457
Received 5 December 2019; Received in revised form 27 February 2020; Accepted 29 March 2020
0011-9164/ © 2020 Elsevier B.V. All rights reserved.
L. Jiang, et al. Desalination 485 (2020) 114457

resistant, theoretically 100% salt rejection, and a higher water recovery

Nomenclature [5–7].
Membrane fouling is a significant problem after long-time operation
For the sample names: [8,9], and biofouling is a particular fouling type in MD systems since its
Raw: raw water samples from the Yellow Sea process is likely to be limited by the operating temperature, which is
NC: samples from the non-concentrating mode known as thermal-effect [10]. With the extension of operation time, the
C: samples from the concentrating mode salinity in feed solution keeps increasing, and the hypertonic environ-
B: samples from the bulk phase ment may also limit the formation of biofouling. However, once bio-
M: samples from the membrane surface films were developed by thermophilic or halotolerant bacteria, the
significant blockage of membrane pores would occur, causing the de-
crease of heat transfer efficiency and sharp flux drop [11,12]. Ad-
raw water from the Yellow Sea raw
ditionally, Bogler and Bar-Zeev [13] found that biofouling can also
non-concentrating mode membrane samples 5d-NC-M
9d-NC-M induce membrane wetting, leading to the contamination of permeates.
14d-NC-M Considering the bio-fouled membrane is hard to recover once bio-
bulk phase samples 5d-NC-B fouling was developed [14], the investigation of the biofouling beha-
9d-NC-B vior in MD process is quite necessary. Researchers [15,16] have studied
14d-NC-B
concentrating mode membrane samples 5d-C-M
the effect of temperature regime, hydrodynamic conditions and water
9d-C-M quality on biofouling, and these experiments were focused on the re-
14d-C-M lationship between operation conditions and permeate flux, while ne-
bulk phase samples 5d-C-B glected the effect of microbial community in natural seawater. Zodrow
9d-C-B
et al. [17] investigated the microbial communities in a closed-loop MD
14d-C-B
system (constant salinity), revealed a shift of microbial community
structure, and proposed the biofilm develop pattern in this system. Liu
et al. [10] investigated the biofouling formation using natural fresh-
water, and devoted efforts on the effect of feed temperature. However,
1. Introduction the change of feed salinity was not evident in these experiments, and
the concentrating-effect on biofouling needs further study.
The lack of clean water is one of the most pervasive problems As MD can cause a significant high feed salinity in the long-term
throughout the world, and seawater desalination is an alternative process, the concentrating-effect should not be neglected during the
method to increase freshwater supply beyond what is available from the investigation. In this study, experiments were operated in non-con-
hydrological cycle [1,2]. One of the most widely used seawater desa- centrating and concentrating modes in parallel to investigate the
lination process was reverse osmosis (RO) followed by multi-stage flash thermal-effect and concentrating-effect. The foulant composition,
distillation (MSF) [3]. However, such process had very high technical membrane morphology, and permeate flux were monitored, and the
complexity and cannot easily be scaled down to very small systems and succession of microbial communities in fouling layer were discussed,
water demands, which limited the application in remote and backward simultaneously, this work emphasized on the interaction between
area [4]. Membrane distillation (MD) is a promising seawater desali- concentrating process and biofouling formation. Finally, the pattern of
nation technology for remote area, as it demonstrates several distinct biofouling formation was proposed, which may assist the improvement
advantages, including a lower required operating temperature com- of MD process by revealing the critical fouling stage.
pared to conventional thermal desalination processes, a lower operating
pressure, a higher energy efficiency, a better performance of fouling-

Fig. 1. Set up of lab-scale DCMD system.

2
L. Jiang, et al.
2. Material and methods
2.1. Experiment setup
A lab-scale DCMD system (Fig. 1) was composed of a DCMD module,
a feed tank, a storage tank, a permeate tank, a balance, a water-bath, a
chiller, two peristaltic pumps, a dosing pump, and a level controller.
The DCMD systems were divided into two operation modes according to
the solutions in the storage tanks, as the tank in concentrating mode
was filled with seawater (prefiltration with 0.22 μm membrane), while
deionized water was used in the non-concentrating mode. The solutions
in the storage tanks were pumped to the feed tank by the dosing pump
(GOSO technology, China) when the level of feed solution was lower
than the level controller. Additionally, a digital camera (POINTGREY
BFLY-PGE-13S2C-CS, Canada) was located above the DCMD module in
the concentrating mode to monitor the fouling process.
During the MD operation, feed and permeate were cycled by the
peristaltic pumps (Longer BT100–2, China) with the cross-flow rate of
10.5 mm/s, and the temperature of water-bath and chiller was set at
50 °C and 10 °C, respectively. Hydrophobic flat-sheet polytetra-
fluoroethylene (PTFE) membrane (Taoyuan Medical and Chemical
Factory, China), with nominal pore size, porosity and effective areas of
0.22 μm, 75–80% and 50 cm2, respectively, was assembled in DCMD
module.
2.2. Water collection
Surface water from the coast of the Yellow Sea near Xiaoyangkou
Harbor (32°31′23.2968″ N, 121°9′54.3384″ E, supplementary SI-1) was
collected. Seawater was pretreated using a mesh film with a pore size of
50 μm to remove the suspended particles, and then was filled in the feed
tank within 12 h.
2.3. Analytical methods
Water flux was measured using a balance, and recorded by the
computer. To avoid the systematic error, the flux data were presented
in terms of normalized flux (NF) J/J0. Where, J and J0 are the in-
stantaneous flux and initial flux, respectively. The theoretical normal-
ized flux, that only accounting for the increase of salinity, was calcu-
lated according to supplementary SI-2, and the baseline test was
conducted with deionized water.
pH and conductivity of feed solution and permeate were measured
using a pH meter (DENVER INSTRUMENT, America) and a conductivity
meter (DDS-11A, China). To monitor the change of dissolved organic
matters in the feed solution, Hitachi F-7000 fluorescence spectro-
photometer (Hitachi Inc., Japan) was used to conduct the three-di-
mensional excitation-emission matrix (3D-EEM) analysis. The mor-
phology of virgin and fouled membranes was investigated by scanning
electron microscope (SEM, FEI Inspect F50, America). X-Ray diffraction
(XRD, Bruker D8 ADVANCE, Germany) and energy dispersive spectro-
meter (EDS, Bruker Xflash6130, Germany) was used to analyze the
component of fouled membrane. To further characterize the biofilms on
the membrane surface, fouled membranes were stained with a mixed
solution of SYTO 9 and propidium iodide (PI) according to the manual
(Filmtracer LIVE/DEAD Biofilm Viability Kit, Invitrogen) and observed
by a fluorescence microscope (Nikon Eclipse 80i, Japan) with the
Excitation/Emission (nm) of 485/498 and 535/617 for live cells and
dead cells, respectively.
2.4. Amplicon sequencing, data processing and statistical analysis
Samples from raw water were filtered through 0.22 μm Durapore
membrane (Millipore GVWP), and together with the membrane sam-
ples, frozen at −80 °C until extraction analysis. DNA was extracted
using the E.Z.N.A.® soil DNA Kit (Omega Bio-tek, Norcross, GA, U.S.)
3
Desalination 485 (2020) 114457
according to manufacturer's protocols, and were amplified by PCR
(TransStart Fastpfu DNA Polymerase, TransGen AP221–02, TransGen
Biotech, China) using primer F338 (5′–ACTCCTACGGGAGGCAGCA–3′)
and primer R806 (5′–GGACTACVSGGGTATCTAAT–3′). Then V3-V4
region of 16S rRNA was adopted to measure the gene on the Illumina
MiSeq platform according to the standard protocols. Sequences found in
this study were submitted to the SRA database (http://www.ncbi.nlm.
nih.gov) under the project accession number SRP232932.
Sequences analysis was performed by UPARSE software package
using the UPARSE-OTU and UPARSE-OTUref algorithms. Graphical
representations of the relative abundance of the bacterial community
were visualized using Krona chart. The circus analysis was conducted
using Circos-0.67-7 (http://circos.ca/), and the Fisher's exact test and
the canonical correspondence analysis (CCA) was performed in R
Studio version 3.2.3 (http://www.r-project.org) using the packages
vegan.
3. Results and discussion
3.1. Characterization of DCMD system
3.1.1. Fouling layer on the membrane surface
To characterize the foulants on the membrane surface at a macro-
scopic level, a digital camera was used in concentrating mode as shown
in Fig. S2. It can be seen that some organic foulants were clearly de-
tected before 8d, and then crystals of CaCO3 (characterized by XRD,
Fig. S3) were gradually appeared.
To monitor the fouling process at a microcosmic level, morphologies
of virgin and fouled membranes were characterized by SEM (Fig. 2B–G
and Fig. S4–6). For the sample 5d-NC-M and 5d-C-M, the surface
morphologies were quite similar, and the membrane was slightly fouled
by the conditioning film, which was mainly formed by dissolved or-
ganics and colloids [18]. In the concentrating mode, with the extension
of operation time, the bacteria subsequently adhered to the con-
ditioning film, forming a mixed fouling layer of organics and early
colonizing bacteria, which called initial biofilm. Additionally, CaCO3
can be observed mostly on the initial biofilm which means deposition of
crystals occurs more readily in the presence of biofouling layer. This
might be due to the higher hydrophilicity of biofilm [19], which led to
the lower critical activation energy for nucleation on the initial biofilm
than that on the virgin membrane, on the basis of the formula in sup-
plementary SI-4. At 14d in concentrating mode, the membrane was
completely covered by CaCO3, and the thickness of mixed fouling layer
was 83.9 ± 9.5 μm. Comparatively, the morphology of membrane in
the non-concentrating mode was significantly different at 9 and 14d,
wherein CaCO3 was not observed. With the extension of operation time,
the coverage of foulants kept increasing, and the mature biofilm, as the
aggregates of microorganisms in which various bacteria were em-
bedded in a self-produced matrix of EPS [20], had completely covered
the PTFE membrane at 14d. Additionally, the cross-section image
showed that the thickness of fouling layer for sample 9d-NC-M and 14d-
NC-M was 13.5 ± 2.1 μm and 20.3 ± 2.4 μm, respectively, indicating
a developing progress of biofouling layer.
To further characterize the biofilms, the fouled membranes were
stained by SYTO9/PI and observed using a fluorescent microscope. The
distribution of live (green) and dead bacteria (red) after 9 days op-
eration was shown in Fig. 2H–I, indicating obvious occurrence of bio-
fouling. The image also showed that there was a higher coverage of
bacteria in non-concentrating mode (89.2%) compared with the con-
centrating mode (57.5%), as the stress from the hypertonic environ-
ment might inhibit the growth of bacteria in concentrating mode. After
14 days operation, the membrane in non-concentrating mode was al-
most completely covered by biofilms (shown in Fig. S7), while the
biofilm cannot be observed clearly in concentrating mode due to the
heavy loading of CaCO3 crystals.
 L. Jiang, et al.

4
Fig. 2. (A) Water flux during MD operation; SEM image of sample (B) 9d-C-M, (C) 14d-C-M, (D) 14d-C-M (cross-section), (E) 9d-NC-M, (F) 14d-NC-M, (G) 14d-NC-M (cross-section); Image of Live/Dead cells in (H) 9d-NC-
M and (I) 9d-C-M. EEM fluorescence spectra of (J) raw water, (K) 5d-NC-B, (L) 5d-C-B, (M) 14d-NC-B, (N) 14d-C-B.
Desalination 485 (2020) 114457
L. Jiang, et al.
3.1.2. Water quality of the bulk phase
To monitor the water quality during the MD operation, measure-
ments of feed conductivity, pH, dissolved fluorescent substances were
performed. As shown in Fig. S9, the feed conductivity in the con-
centrating mode increased with the operation time from 40.60 to
104.98 mS/cm, while this value stabilized around 40.50 mS/cm in the
non-concentrating mode. The pH also showed different trends, as it
stabilized around 8.0 in non-concentrating mode, while a drop from
8.12 to 7.36 was observed after 9d in concentrating mode. The drop of
pH was correlated to the crystallization process of CaCO3, and will be
discussed in supplementary SI-5.
To analyze the chemical composition of fluorescent substance in
feed solution, 3D-EEM analysis was conducted, and the contour was
subdivided as shown in Fig. 2J–N, wherein the regions I, II, III, IV, V,
and VI were ascribed to the tyrosine, tyrosine-like proteins, tryptophan,
tryptophan-like proteins, fulvic acid-like, and humic acid-like sub-
stances, respectively [21]. The results showed that tryptophan and
fulvic acid-like substance were the major components in raw water,
while peaks of tryptophan and tryptophan-like proteins became major
substances after 5 days operation. The appearance of new peak and the
intensity increment of fulvic acid-like substance might be attributed to
the release of organics from suspended solids and dead cells, as the
higher temperature in the bulk phase can accelerate this process. Ad-
ditionally, the metabolism of bacteria was another factor [22], and it
was reported that tryptophan-like dissolved organic matters re-
presented products of bacterial activity and bioavailable substrates
[23]. For sample 9d-NC-B and 9d-C-B, the major peaks were still
tryptophan and tryptophan-like proteins as 5d samples, and the in-
tensity of tryptophan-like proteins showed an obvious increase (Shown
in Fig. S10). Considering the analysis of microbial community structure
in Section 3.2.2, hypothesis was proposed that some bacteria in the MD
system, for example Rhodobacteraceae [24], can secrete bioavailable
substrates, leading to the increasingly high intensity of tryptophan-like
proteins, making the environment suitable for the bacteria which re-
quire higher nutrient level. With the extension of operation time, tyr-
osine-like proteins were observed to be the major peak at 14d, which
might be correlated with the high abundance of Hyphomonadaceae, a
bacteria that can release different types of EPS [25].
3.1.3. Permeate flux changes
During the MD experiment, the initial flux was around 9.0 L/m2·h,
and the conductivity of permeate was stabilized around 10 μS/cm with
rejection over 99.9%. The trend of normalized flux in both operation
modes, the theoretical normalized flux in concentrating mode, and
baseline tests are shown in Fig. 2A. In the non-concentrating mode, the
NF displayed slow decline, rapid decline, and moderate decline pro-
gress. During stage I (0–150 h), the NF dropped to 95.2% and the de-
cline rate was 0.02%/h. This was a transitional period between organic
fouling and biofouling, in which membrane surface was mainly con-
taminated by large particles and organic conditioning film. With the
action of permeate drag force and gravity force, nanometer-thin “con-
ditioning films” were considered to be formed in this period, which
mainly consisted of organic foulants in the feed solutions, soluble mi-
crobial products (SMP) and secreted extracellular polymeric substances
(EPS) [26]. According to the 3D-EEM analysis, a new peak of trypto-
phan-like proteins was observed, which might be relevant to SMP and
EPS. Besides, the SEM image showed that only a few bacteria-like
structures were observed in the conditioning film, and EDS mapping
(Fig. S5) also showed that fewer phosphorus was found in samples 5d-
NC-M compared with the samples 9d-NC-M. As phosphorus was the
representative element in nuclear acid and cell membrane, the rela-
tively lower abundance might indicate that bacteria was not an major
component in the foulants at 5d. This phenomenon can be attributed to
the same surface charge of bacteria and PTFE membrane. It was re-
ported that PTFE membrane and most bacteria was negatively charged,
and thus the repulsion force can inhibit the adhesion of bacteria on
5
Desalination 485 (2020) 114457
membrane surface [27,28]. The formation of conditioning film would
enhance the adhesion of organic foulants on the membrane surface,
facilitating the attachment of microorganisms via van der Waals, hy-
drophobic and hydrogen bonding interactions [12], leading to the high
coverage of initial biofilm in stage II. During stage II (150–220 h), there
was a dramatical drop of NF from 95.2% to 74.7% with a decline rate
up to 0.31%/h. In this period, membrane was covered by the fouling
layer with a more compact structure, and the microbial analysis showed
a highest abundance of initial biofilm forming bacteria (for example
Rhodobacteraceae). Considering the morphology of foulants and the
shift of microbial community structure, the flux drop might be mainly
attributed to biofouling. The coverage of foulants (78.7%) was much
higher than that in sample 5d-NC-M (28.1%), and the blockage of
permeate sites resulted the significant flux drop. During stage III
(220–335 h), the NF only dropped from 74.7% to 70.3%, and the de-
cline rate was lower than 0.05%/h. The permeability sites on PTFE
membrane was completely covered with rather stabilized biofouling
layer, and the flux drop might be attributed to the increase of fouling
layer thickness, as the thickness increased from 13.5 ± 2.1 μm to
20.3 ± 2.4 μm (according to SEM cross-section image).
Comparatively, the flux in the concentrating mode experienced a
slow decline, moderate decline and rapid decline progress. During stage
I (0−110 h), the NF dropped to 93.3% with a decline rate of 0.06%/h,
which was slightly higher than the theoretical decline rate (0.02%/h).
This was a transitional period between organic fouling and biofouling,
which was similar to the non-concentrating mode. In addition to the
effect of organic fouling, the increase of feed salinity was another im-
portant factor which cause flux drop in concentrating mode, as higher
salinity may cause a lower vapor partial pressure. During stage II
(110−210 h), the NF dropped from 93.3% to 78.8%, and the decline
rate (0.12%/h) was much higher than the calculated values (0.02%/h).
Besides the factors of biofouling mentioned in non-concentrating mode,
the higher decline rate also might be resulted from the effect of con-
centration polarization. During the membrane distillation process, with
the accumulation of rejected solutes and particles, a boundary layer was
formed on the membrane surface and this phenomenon was called
concentration polarization [29]. The concentration of NaCl in the po-
larization layer was much higher than that in the bulk phase [30],
leading to a lower vapor pressure, and hence a severe flux drop was
observed. During stage III (210–335 h), the NF significantly dropped
from 78.8% to 47.1% with a decline rate of 0.23%/h, which was 2 times
of that in stage II. The halotolerant bacteria have become dominant
species in this period, and a mixed layer of inorganic scaling and bio-
fouling with the thickness of 83.9 ± 9.5 μm has completely covered
the membrane, suggesting that inorganic scaling and biofouling were
the major factors that deteriorated the membrane performance.
3.2. Analysis of the microbial community in MD system
The characterization of fouled membrane and bulk phase revealed
that the biofouling is an important factor that affected the membrane
performance in both operation modes. To further investigate the for-
mation of biofouling, high throughput sequencing (MiSeq Illumina) was
employed in this experiment. After denoising and subsampling, a total
of 242,411 sequences (18,647 in each sample) were obtained and
clustered into 582 OTUs with 97.0% sequence similarity. The coverage
level of all the samples were higher than 0.99, indicating that the se-
quencing depth could well reflect the microbial community of the
samples.
3.2.1. The thermal-effect and concentrating-effect on the microbial
community diversity
To generally estimate the change of the microbial community, es-
timators of community richness (Chao, Sobs, Ace) and community di-
versity (Shannon, Simpson) were listed in Table S1. The raw water has
the highest Sobs index and Shannon index of 387 and 4.09,
L. Jiang, et al.
Fig. 3. Shannon index (A) and Sobs index (B) of raw water and membrane samples.
respectively, which means most bacteria from the surface water will be
inhibited by the thermal-effect in the MD system, leading to the de-
crease of community richness and diversity. The richness and diversity
displayed different changing trend as shown in Fig. 3.
For the Sobs index, the changing trend was quite different between
concentrating and non-concentrating modes. The Sobs index kept de-
creasing from 5 to 14d in the concentrating mode, while that in the non-
concentrating mode fluctuated around 200. This phenomenon can be
attributed to the stress of concentrating-effect, which can inhibit some
bacteria by the increasingly high salinity. Comparatively, the Shannon
index kept increasing with the operation time from 5 to 14d in the
concentrating mode, while the index in non-concentrating mode
dropped to 1.11 at 9d, and then followed by a significant increment to
3.78 at 14d. The change of Shannon index reflected the selective
pressure of temperature and the microbial community succession, as
many bacterial died and was decomposed due to the sudden change of
feed temperature in the initial stage, and the bacteria which can survive
better in MD system was gradually enriched from 5 to 14d, resulting a
higher microbial diversity. Additionally, the lowest Shannon index in
sample 9d-NC-M might be attributed to the significantly high abun-
dance of Rhodobacteraceae which played an important role in the for-
mation of initial biofilm (detailedly discussed in supplementary SI-6).
3.2.2. The shift in microbial community structure
The succession of the microbial community usually plays a key role
in the membrane biofouling process [31], and the bacterial community
compositions at various taxonomic levels (phylum, class, and family)
are shown in Fig. 4A-C. At phylum level, the community structure in
raw water and MD samples displayed significant difference, as over
96% of bacterial sequences was related to six different phyla for the raw
water, including Proteobacteria (56.3%), Bacteroidetes (29.4%), Parcu-
bacteria (5.4%), Actinobacteria (2.2%), Verrucomicrobia (2.1%) and Cy-
anobacteria (1.2%), while over 98% of bacterial sequences was related
to five phyla for the 6 MD samples, which were Proteobacteria
(41.7%–94.6%), Firmicutes (1.9%–37.0%), Bacteroidetes (1.2%–23.4%),
Planctomycetes (0.8%–14.7%) and Chlamydiae (0.1%–2.2%). The dif-
ferent bacteria community structures were resulted from the thermal-
effect, the concentrating-effect, and the biofilm formation and ma-
turation.
Proteobacteria was an important phylum as it took the largest pro-
portion in all the samples, and Alphaproteobacteria was the major class
in this phylum. As shown in Fig. 4B, after an increment from 38.0% to
82.5% on 5d, the abundance of Alphaproteobacteria kept decreasing
until the end of operation in concentrating mode with the abundance of
6
Desalination 485 (2020) 114457
35.7%. In the non-concentrating mode, the abundance of Alphaproteo-
bacteria increased to 89.1% at 9d, then followed by a decrease to 61.5%.
This changing trend might be associated with the biofilm formation in
different stages. Tracing the data to the family level, Rhodobacteraceae
was considered to be an important family in class Alphaproteobacteria,
and it has been reported that Rhodobacteraceae was the key member of
the microbial community for the initial biofilm [32]. Planctomycetes
was another important phylum that enriched in concentrating mode, as
its abundance increased from 0.8% to 14.7% during the 9d operation,
and then followed by a decrease to 8.0% at the end of operation, which
showed similar trend with Alphaproteobacteria. It was reported that
Planctomycetes also played an essential role in early stages of biofilm
formation, as Planctomycetes carried genes related to pili and flagella
[33]. Compared with Proteobacteria and Planctomycetes, the phylum
Bacteroidetes showed an opposite trend, as the abundance of Bacter-
oidetes decreased from 29.4% to less than 6.7%, and finally recovered to
13.0% and 23.4% at 14 d for concentrating and non-concentrating
modes, respectively. This trend was resulted from the formation of
mature biofilm [32].
Combined with the Circos analysis, unclassified_f_Rhodobacteraceae
and SM1A02, which belonged to Proteobacteria and Planctomycetes, re-
spectively, played important roles in the formation of initial biofilm,
while Bacteroidetes was essential for biofilm maturation. Additionally,
some Proteobacteria, for example Hyphomonas and henriciella were
predominant genus after 14d operation, because they played important
roles in the maturation of biofilm, and can produce different types of
EPS, which enhanced the adhesion of surface and is helpful for the
formation of biofilm matrix [34,35].
Firmicutes was another noteworthy phylum, as Firmicutes was a
dominant bacteria in most MD samples with the proportion of
1.9%–37.0% compared to the raw water of less than 0.1%. This phe-
nomenon was mainly caused by the higher temperature of 50 °C com-
pared to the 10 °C raw water, and researchers have found that Firmicutes
was usually a dominant bacteria in a hot environment, such as hot
spring [36,37]. At the end of operation, a significant higher abundance
of Firmicutes was observed in the concentrating mode with the pro-
portion of 37.0%, which was much higher than it in non-concentrating
mode (2.2%). Tracing the reasons to the genus, almost all the Firmicutes
were Bacillus, which can form spores which was a cell type that has a
great resistance to heat, hypertonic, alkali, acid and radiation [38],
leading to a survival advantage in the concentrating mode. At species
level, alveayuensis took the proportion of over 99% in Bacillus, and it
was reported that alveayuensis can grow in the environment with up to
13% (wt/vol) NaCl and 80 °C [39].
L. Jiang, et al.
Fig. 4. Microbial community structure at (A) Phylum (B) Class (C) Family level, and (D) Circos analysis at genus level.
3.2.3. The Fisher's exact test and the environmental variables analysis
To further characterize the differences between non-concentrating
and concentrating modes in each phase, Fisher's exact test was per-
formed based on DP: Asymptotic. As shown in Fig. 5A–C, the bacteria
with most significant difference in concentrating mode after 14d op-
eration was Bacillus. The difference between proportions of the Un-
classified_f_Rhodobacteraceae was 13.3% and 29.9% at 5d and 9d, re-
spectively, while this value decreased to −1.1% at 14d. At the end of
operation, the difference between proportions of Hyphomonas and
Ponticaulis was 10.5% and 7.8%, respectively, while no significant dif-
ference was observed at 5d and 9d. The fisher's exact test revealed that
Unclassified_f_Rhodobacteracea played an important role during the early
stage of biofouling, and survived better in non-concentrating mode.
Hyphomonas and ponticaulis were representative genus in non-con-
centrating mode at the end of operation, while Bacillus was largely
enriched in concentrating mode.
Additionally, CCA was conducted at genus level, to further reveal
the relationship between the variables (operation time and salinity) and
the community succession. As shown in Fig. 5D, acute angles emerged
among operation time and salinity, indicating that these variables had a
synergetic impact on the microbial community. 9 of 28 genus were
positively correlated with concentration, and 18 of 28 genus were po-
sitively correlated with the operation time. The raw water sample and
membrane samples were clustered separately, indicating that the high
temperature took effect within a relatively short time, leading to an
obvious shift of microbial community structure. Comparatively, 5d-C-
M, 5d-NC-M, 9d-C-M and 9d-NC-M, were closely clustered, which
7
Desalination 485 (2020) 114457
means the concentrating-effect can remarkably change the community
structure only when the salinity was high enough. The samples of 14d-
C-M and 14d-NC-M were grouped separately, and showed large dis-
tances from other MD samples, indicating that a significant shift of
microbial community structure was accompanied with the maturation
of biofilm in a short time.
3.3. The fouling pattern of the MD desalination process
In this study, non-concentrating mode was influenced by the high
temperature, while the combined effect of temperature and salinity
occurred in concentrating mode. The comparison between non-con-
centrating mode and raw water may reveal the thermal-effect, and the
comparison between two operation modes was also need to distinguish
concentrating-effect from thermal-effect. The influence of temperature
is mainly manifested in the selection of thermophilic bacteria, as it was
hard for other bacteria to survive in the environment of over to 50 °C,
and thus dead in a relatively short time. Meanwhile, the high tem-
perature might also induce the release of organics from suspended so-
lids and dead cells, and accelerating the formation of conditioning film.
The concentrating-effect was typically reflected in three aspects, in-
cluding the occurrence of concentrating polarization, the formation of
inorganic scaling, and the selection of halotolerant bacteria.
Concentrating mode was commonly used in seawater desalination, and
its fouling behavior was more complex than non-concentrating mode.
As shown in Fig. 6, to propose the fouling process during seawater MD
desalination clearly, it was better to reveal the fouling pattern in non-
L. Jiang, et al.
Fig. 5. Fisher's exact test bar plot on Genus level for samples after (A) 5d, (B) 9d and (C) 14d operation; (D) CCA analysis of microbial community and environmental
variables.
concentrating mode firstly, and then taking the concentrating-effect
into consideration.
For the non-concentrating mode, the fouling process can be divided
into 3 stages. Phase I: The formation of conditioning film and thermal-
effect. During this period, the suspended particles, colloids and dis-
solved organic matters would attach to the membrane surface and
microorganism might reach the membrane with the suspended solids or
colloids. The dissolved organic foulants and secreted SMP/EPS might
accumulate on the membrane surface under the action of permeate drag
force and gravity force, forming a nanometer-thin “conditioning film”
[18,26]. Commonly, the conditioning film can enhance the attachment
of bacteria, showing a better adhesion than virgin PTFE membrane
[12], leading to the formation of initial biofilm in Phase II. Ad-
ditionally, the sudden change of temperature from 10 to 50 °C might
inhibit the bacteria, for example NS3a_marine_group and Sulfitobacter,
resulting a sharp decrease of bacteria diversity. Phase II: The formation
of initial biofilm. In this stage, the attachment of some bacteria on the
membrane was occurred, but the environment on the membrane sur-
face cannot support the growth of most bacteria, as these bacteria (for
example Bacteroidetes) require a relatively higher level of organics and
nutrients [40]. Comparatively, Rhodobacteraceae was positively selected
in the low nutrient environment [41], and played an important role in
the formation of initial biofilm via the secretion of extracellular factors
[24]. During this period, the permeability sites were covered by the
8
Desalination 485 (2020) 114457
rapid growing initial biofilm, leading to a significant flux drop. Phase
III: Biofilm maturation. With the biofilm formation and metabolism, the
living condition was gradually changed, and some bacteria in the bio-
film, for example Hyphomonadaceae, showed a significant increase of
abundance. These bacteria can release different types of EPS, and
therefore the structure of mature biofilm is thicker and more compact
than the initial biofilm [34]. Moreover, the dense structure might lead
to a temperature polarization and protect the bacteria in the biofilm
from the hot environment in bulk phase. In such conditions, the mature
biofilm which contained various bacteria was gradually formed, leading
to a lower abundance of Rhodobacteraceae and a higher diversity of
microbial community.
Considering the fouling pattern in non-concentrating mode, the
concentrating mode can also be divided into 3 stages. Phase I: The
formation of conditioning film and thermal-effect. As the salinity was
not very high during this period, the fouling process was similar to the
non-concentrating mode. Phase II: The formation of the initial biofilm
and initial scaling. In this stage, some Alphaproteobacteria early colo-
nized on the membrane, and gradually formed the initial biofilm with
other bacterial, such as Planctomycetes which was a phylum only greatly
enriched in concentrating mode. Additionally, inorganic foulants were
gradually precipitated on the membrane surface, especially on the
biofouling layer, as the foulants have higher hydrophilia than virgin
membrane, leading to a lower critical activation energy for nucleation
 L. Jiang, et al.

9
Fig. 6. Two fouling patterns during MD operation.
Desalination 485 (2020) 114457
L. Jiang, et al.
[19]. Phase III: Concentrating-effect and severe scaling. During the final
phase, biofilm was further developed and large amount of inorganic
scaling were formed on the membrane surface. The microbial com-
munity structure was quite different from non-concentrating mode, as
halotolerant bacteria was positively selected in concentrating mode due
to the increase of salinity and the effect of concentration polarization.
Additionally, the inorganic scaling would greatly increase the thickness
of fouling layer, and the secreted EPS from mature biofilm will fill the
interstice among crystals, resulting in a significant flux drop.
4. Conclusion
This research devoted efforts to characterize the fouling process of
DCMD system using seawater, with an especial focus on the interaction
between concentrating process and biofouling. The fouling behavior
was systematically analyzed and the results led to the following con-
clusions:
(1) The flux decline in MD system was mainly attributed to the mem-
brane scaling and biofouling. Although the high temperature and
salinity might inhibit some bacteria during MD process, biofouling
still exist after a long-term operation, and might accelerate the
formation of inorganic scaling.
(2) Some thermophilic and halotolerant bacteria, for example Bacillus,
can be positively selected by the hot and hypertonic environment,
and these bacteria would cause significant flux drop.
(3) Some Proteobacteria, for example Rhodobacteraceae, played an im-
portant role in the development of initial biofilm, while
Bacteroidetes contributed to the maturation of biofilm. Additionally,
Hyphomonadaceae which belonged to Proteobacteria, were enriched
in the later phase of MD operation, and was related to formation of
mature biofilm matrix.
(4) Increasing operating temperature and salinity still has limitations
when developing anti-biofouling MD technology due to the tem-
perature polarization and microbial community succession, which
should be focus in further research.
CRediT authorship contribution statement
Longjie Jiang:Conceptualization, Formal analysis, Investigation,
Writing - original draft.Lin Chen:Funding acquisition, Writing - re-
view & editing, Methodology.Liang Zhu:Funding acquisition,
Writing - review & editing.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgements
This study was mainly financially supported by National Natural
Science Foundation of China (grant number: 51508153), Fundamental
Research Funds for the Central Universities (grant number:
B200202107) and a project funded by the Priority Academic Program
Development of Jiangsu Higher Education Institutions (grant number:
BK20150813).
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.desal.2020.114457.
10
Desalination 485 (2020) 114457
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