Journal Pre-proofs

Black odorous water concentrating by forward osmosis (FO) with aquaporin

biomimetic membranes: pollutants concentrating and membrane fouling char‐
acteristics

Yong Zhang, Kai-Tong Wang, Wei-Li Jiang, Jia-Ying He, Hong Wang, Bing
Li, Ming Gao

PII: S1385-8947(20)32066-0
DOI: https://doi.org/10.1016/j.cej.2020.125938
Reference: CEJ 125938

To appear in: Chemical Engineering Journal

Received Date: 11 April 2020

Revised Date: 13 June 2020
Accepted Date: 16 June 2020

Please cite this article as: Y. Zhang, K-T. Wang, W-L. Jiang, J-Y. He, H. Wang, B. Li, M. Gao, Black odorous
water concentrating by forward osmosis (FO) with aquaporin biomimetic membranes: pollutants concentrating
and membrane fouling characteristics, Chemical Engineering Journal (2020), doi: https://doi.org/10.1016/j.cej.
2020.125938

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Black odorous water concentrating by forward osmosis (FO) with

aquaporin biomimetic membranes: pollutants concentrating and

membrane fouling characteristics

Yong Zhang a,b,d,e,f, Kai-Tong Wang a, Wei-Li Jiang b, Jia-Ying He a, Hong Wang a,*, Bing Li c,

Ming Gao c,**

a
School of Environment, Nanjing Normal University, Nanjing 210023, China

b
Jiangsu Provincial Key Laboratory of Environmental Engineering, Nanjing 210000, China

c
Jiangsu Academy of Environmental Industry and Technology, Nanjing 210000, China

d
Key Laboratory of Virtual Geographic Environment (Nanjing Normal University), Ministry

of Education, Nanjing 210023, China

e
State Key Laboratory Cultivation Base of Geographical Environment Evolution (Jiangsu

Province), Nanjing 210023, China

f
Jiangsu Center for Collaborative Innovation in Geographical Information Resource

Development and Application, Nanjing 210023, China

* Corresponding author. Contact address: School of Environment, Nanjing Normal University,

1 Wenyuan Road, Nanjing 210023, China. Tel: +86 25 85891490.

** Corresponding author.

E-mail address: hongwang@njnu.edu.cn (Wang H.); shy_minggao@163.com (Gao M.).

1
Abstract: This study aimed to investigate pollutant concentration and membrane fouling

characteristics of a forward osmosis (FO) process using aquaporin biomimetic membrane

(ABM) for concentrating black odorous water. The membrane cells were operated in active

layer facing feed solution (ALFS) mode with 2 M NaCl solution as the draw solution. The

system was continuously performed for 64 batch cycles, and each cycle duration was 24 h. At

the end of each cycle, physical cleaning with deionized water was employed as the membrane

recovery strategy. The results showed that the rejection ratios of chemical oxygen demand

(COD), total phosphorus (TP) and nitrate (NO3--N) could reached 97.2%, 98.0%, and 85.0%,

respectively, while most NH4+-N penetrated into the draw solution due to cation exchange.

The total nitrogen (TN) rejection ratio was largely dependent upon the NH4+-N/TN ratio. At

high NH4+-N/TN ratio, the successions of ammonia oxidizing bacteria (AOB) communities

enriched in the biofouling layer in different experimental stages would affect the

transformation degree of NH4+-N, and thus lead to much fluctuations of TN rejection. The

average initial water flux reached 9.84 L/(m2·h), and the average water flux of each cycle kept

stable especially in the later stage of the experiment. In the biofouling layer, polysaccharides

enhanced while proteins decreased in the later period. P, Mn, Fe and Na also accumulated on

the surface. Norank_f__Reyranellaceae, Erythrobacter, SM1A02, Pirellula, and

Hydrogenophaga were the predominant genera enriched in the fouling layer, which would

lead to complex pollutants transformations, especially nitrogen transformation, during the FO

process.

Keywords: Black odorous water; Forward osmosis; Aquaporin biomimetic membrane

(ABM); Water flux; Nitrogen; Bacteria community

2
1. Introduction

Black odorous water is a typical pollution phenomenon which always occurred in urban

water bodies caused by human activity, domestic and untreated industrial wastewater drainage,

agricultural runoff, etc. [1]. Black odorous water has been reported in many developed and

developing countries [2], where both the water resource security and the water environment

quality were severely threatened. In China, nearly 2,000 water bodies were reported to be

classified as black odorous water by the end of February 2017, in which rivers accounted for

85.7% and lakes accounted for 14.3%, respectively [3]. Several physical, chemical and

biological methods have been proposed to remediate the severely polluted surface waters, e.

g., aeration [4], coagulant dosing [5], chemical inactivation [6]. However, these methods

attempted to control the pollution by means of changing the chemical forms of the pollutants,

and the secondary pollution is a common problem for these methods. Sediment dredging,

even extensively used in China, is still contradictory whether it is a suitable treatment for

black odorous water [7]. Although ecological restoration is an effective and environmental

friendly method for improving the structure and the function of the river ecosystem [8], it is

difficult for rapid restoration of the black odorous water.

Forward osmosis (FO), an emerging membrane technology, has attracted considerable

attention in recent years [9]. Compared to conventional pressure driven membrane processes,

FO employs osmotic pressure as the driving force of the membrane filtration, with the result

of many advantages, including low energy consumption, low membrane fouling propensity,

and high rejection of pollutants, etc. [10]. Dozens of literatures have compared the

performance of the FO membrane materials, e. g., the asymmetric cellulose triacetate (CTA)

3
membrane and the thin film composite (TFC) membrane [11-13]. Recently, the aquaporin

biomimetic membrane (ABM) has attracted attentions due to its advantages of high water

permeability and excellent solute selectivity [14,15]. FO has been reported to be employed in

many kinds of wastewater treatments. Generally, one important original intention of FO

application in wastewater is to concentrate the pollutant to a higher concentration for more

effective treatment and/or energy recovery, such as anaerobic digestion of low-strength

wastewater [16] and even greywater concentration by FO-pretreatment [17]. Although the

water indexes of the black odorous water could be temporarily improved by changing the

chemical forms of the pollutants via the conventional methods above mentioned, thoroughly

removal of these pollutants from the water body is of long-term significance. Therefore, an

efficient concentration of the polluted water to a considerable small volume is urgent for

further effective treatments. However, little work has been conducted about the feasibility of

concentrating the black odorous water by FO.

Pollutant leakage and membrane fouling are always concerned by researchers in FO

process [18,19]. Previous studies have reported the leakage of small organic compounds,

ammonium, and phosphate from the feed solution to the draw solution [10]. The pollutant

layer is consisted of inorganic/organic foulants and microorganisms presents a gradual

increasing trend on the FO membrane surface with the extension of operation time [20].

Different wastewaters would lead to different fouling degree of membrane. For example,

Volpin et al. [19] reported that under the same conditions, the flux decline degree was

negligible, mild and severe with the feed solution of the FO process being secondary effluent,

biologically treated wastewater and primary effluent, respectively. Biofouling mitigation

4
methods include rinsing or flushing, air-sparging, cleaning with different chemicals [21], etc.

It is noteworthy that the adsorption of some metal ions on the FO membrane surface would

also lead to membrane fouling [22]. Regarding the characteristics of the black odorous water,

and also for the convenience of membrane fouling study, this work employed a single

physical flushing with deionized water at the end of each cycle as the membrane recovery

strategy.

The objective of this study was to investigate the feasibility of the FO technology on the

concentration of the black odorous water. Lab-scale FO systems using ABM were constructed

for real black odorous water treatment. The concentrating effects of the organic matters

denoted by chemical oxygen demand (COD), nitrogen, total phosphorus (TP) were observed,

and the membrane fouling was analyzed by the flux reduction combined with

micromorphology changing of the membrane surface. The dynamic changes of the structures

of the ammonia-oxidizing bacteria (AOB) community and the total bacterial community in

membrane fouling layer were also investigated by Illumina sequencing technology. This work

intended to serve as a reference for the feasibility of FO technology in the effective treatment

of the black odorous water.

2. Materials and methods

2.1. Experimental materials

The black odorous water was taken from a eutrophic pond located in Nanjing, China with

the concentrations of the main indices as follows: 7.33-7.70 of pH, 23-60 mg/L of COD,

4.4-8.2 mg/L of total nitrogen (TN), 2.1-4.3 mg/L of ammonia nitrogen (NH4+-N), and

5
0.19-0.73 mg/L of TP.

2.2. Reactors and experimental design

The experiments were carried out in four identical bench-scale cross-flow membrane

cells running in active layer facing feed solution (ALFS) mode. Each cell is comprised of two

symmetric flow channels which were separated by the ABM with the effective area of 50 cm2.

The ABM was bought from Aquaporin A/S located in Kongens Lyngby, Denmark. The

membrane specifications provided by the producers could be found in Table S1 in the

supplementary materials. The size of the channel was 100 mm in length, 50 mm in width and

3 mm in depth. All experiments were conducted in batch mode. Initially, 1L black odorous

water was used as feed solution and 1 L 2 M NaCl as draw solution. The two solutions were

kept circulating at the flow rate of 0.6 L/min through the FO membrane by two variable-speed

gear pumps (Runze fluid control equipment Co., Ltd, Nanjing, China). With the operation of

the device, the increasing quantity of the draw solution was weighted by a digital balance and

regularly recorded by a computer for water flux calculation. With the water permeating from

the feed side to the draw side, the black odorous water was concentrated while the draw

solution was diluted, leading to gradual decline of the water flux. At the end of the cycle, the

feed and draw solutions were both replaced by deionized water for physical cleaning of the

ABM with the flushing speed of 1.5 L/min for 30 min. Afterwards, the device was reset for

the next cycle. The system was performed for 64 cycles with each cycle of 24 h. The

experimental temperature was controlled at 23-25℃. The concentrated black odorous water

and the diluted draw solution were collected at the end of each cycle for further determination,

6
and the membrane samples at the end of the 18th, the 36th, the 48th and the 64th cycles were

taken for membrane fouling analysis.

2.3. Analytical methods

COD, BOD5 (biochemical oxygen demand), NH4+-N, NO3--N, TN and TP were assayed

according to the standard methods [23]. The water flux (Jw) in this research was calculated

according to Eq. (1). The average water flux ( ) of each cycle was calculated using Eq. (2):

(1)

(2)

Where, ΔV and ΔVT are the increasing volumes of the water at time T=Δt and T=24h,

respectively. A is the effective area of the ABM, and Δt is the time interval between two

adjacent recorded quality of the draw solution (30 min).

Concentration multiple (N) of each cycle was calculated using Eq. (3), and rejection ratio of

pollutants (R) was calculated using Eq. (4):

(3)

(4)

Where, Ct and C0 are the concentrations of the pollutants at time t=t and t=0, respectively.

Vt and V0 are the volumes of the feed solution at time t=t and t=0, respectively.

T-test was conducted to analyze the significance of the results and p＜0.05 was considered

to be statistically significant.

7
2.4. Membrane fouling analysis

2.4.1 Morphological analysis of membrane fouling layer

The confocal laser scanning microscopy (CLSM, MRC-1024, BIO-RAD Co., USA)

observation about the proteins and α-d-glucopyranose polysaccharides in the membrane

fouling layer was conducted after the membrane surface was sequentially stained with

fluorescein isothiocyanate (FITC) (Molecular Probes, Eugene, OR) and Concanavalin A (Con

A, Molecular Probes, Eugene, OR). The staining method was according to the method

reported by Chen et al. [24].

Microscopic membrane surface morphology was observed by a scanning electron

microscope (JSM-7600F, Japan). Dynamic changes of the elemental compositions of the

membrane fouling layer during the experimental process were detected by an X-ray

energy-dispersive spectrometry (Oxford, INCA X-act, UK) [25].

2.4.2 Microbial population diversity of biofouling layer

Four identical membrane cells were parallel running for regular collection of samples of

the fouling layer on the membrane surface for surface morphology and bacterial sequencing

analysis, as shown in Fig. S1. "M1" was the sample taken at the end of the 16th cycle, "M2"

of the 32nd cycle, "M3" of the 48th cycle, and "M4" of the 64th cycle. For comparison, 1 L of

black odorous water was filtered with 0.45 μm membrane and the filter cake was taken as the

sample to represent the microbial community in the influent, named "M0".

DNA was extracted from the five samples using the E.Z.N.A.® soil DNA Kit (Omega

Bio-tek, Norcross, GA, U.S.) according to manufacturer’s protocols. Two sets of primers were

designed for further PCR amplifications, e. g., primers cd3aF

8
(5'-GTSAACGTSAAGGARACSGG-3') and R3cd (5'-GASTTCGGRTGSGTCTTGA-3') to

investigate the bacterial structure [26], and primers bamoA1F

(5'-GGGGTTTCTACTGGTGGT-3') and bamoA2R (5'-CCCCTCKGSAAAGCCTTCTTC-3')

for AOB community analysis [27].

PCR procedure was: 95 °C for 3 min, followed by 32 cycles of 30 s at 95 °C, 30 s at 55 °C,

45 s at 72 °C and a final extension of 10 min at 72 °C. PCR reactions were performed in 20

μL mixture containing 4 μL of 5 × FastPfu Buffer, 2 μL of 2.5 mM dNTPs, 0.8 μL of each

primer (5 μM), 0.4 μL of FastPfu Polymerase, 0.2μL of BSA and 10 ng of template DNA.

Amplicons were sequenced on the Illumina MiSeq platform (PE300) at Shanghai Majorbio

Bio-Pharm Technology Co., Ltd (Shanghai, China). The phylogenetic affiliation of each 16S

rRNA gene sequence was analyzed by RDP Classifier (http://rdp.cme.msu.edu/) against the

silva (SSU115) 16S rRNA database with a set confidence threshold of 70% [28].

The raw reads have been deposited into the National Center for Biotechnology

Information (NCBI) Sequence Read Archive (SRA) database. The accession numbers for

bacterial 16S rRNA gene sequences are SRR10418893 - SRR10418897, and for AOB

functional gene are SRR10418773 - SRR10418777.

3. Results and discussion

3.1. Pollutant concentration

The dynamic changes of COD, TN and TP of the feed solutions with the increasing of the

concentration multiple are shown in Fig. 1. The red lines in the figure represent the theoretical

values when the rejection ratios of the pollutants are 100%. The average COD rejection ratio

9
could reach 97.2%, and it kept high at every concentration multiple. The results were

consistent with many former studies reporting that FO membrane could effectively intercept

organic matters in wastewater, especially macromolecular organics [13,15,29,30]. Madsen et

al. [31] even found that the aquaporin membrane has significantly higher rejection values for

three trace organics than the cellulose acetate membrane. A variety of organic contaminants

contribute to the organic pollution of the black odorous water. For example, an investigation

about the microbial functional gene by Xu et al. [32] showed that most detected genes were

involved in carbon compound degradation, including starch, pectin, hemicelluloses, cellulose,

aromatic, chitin and lignin, etc. Liang et al. [33] reported that the biorecalcitrant humus,

resistant to microbial degradation, accounts for over 40% of total organic matter in urban

rivers. The biodegradability of the black odorous water before and after concentrating was

further determined by the ratio of BOD5 and COD (B/C), as shown in supplementary Fig. S2.

No significant difference of the biodegradability occurred (p＞0.05), and the average B/C

ratios of the water before and after FO concentrating were 0.26 and 0.27, respectively. The

results indicated that even FO could concentrate the black odorous water to a relative high

COD concentration, anoxic hydrolysis or other pretreatments should be necessary to enhance

the biodegradability for further biological treatment.

(Fig. 1)

The changes of TP concentration with the concentrating multiple presented the similar

laws as COD (Fig. 1(c)). The rejection ratio kept high during the whole process, and the

10
average rejection ratio reached as high as 98.0% (Fig. 1(d)). Actually, good retention of

phosphate and TP has been demonstrated in integrated FO-biological processes due to the

excellent retention ability of the FO membrane [30]. Melin et al. [34] also reported an

outstanding high rejection ratio of phosphorus compounds of 99.7% by FO. Consequently,

effective phosphate recovery based on FO technology has also been proposed by some other

researchers [35].

The rejection ratio of TN was significantly lower than those of COD and TP (Fig. 1 (b)).

Furthermore, statistical results (Fig. 1 (d)) show that the TN rejection ratios fluctuated

violently (25.4%-88.0%) during the 64-cycle experimental duration. Dozens of literatures

have reported the nitrogen rejection characteristics of FO membrane, most of which focused

on CTA and TFC membranes. For example, Zou et al. [36] employed commercial all-purpose

fertilizers as draw solution, and found that about 50% of TN diffused through the CTA

membrane from the draw side to the feed side within 24 h. Linares [37] reported a rejection

ratio of TN in scope from 56% to 59% by a CTA membrane. Wang et al. [38] reported that,

when concentrating municipal wastewater by a CTA membrane with 0.5M NaCl solution as

draw solution, the rejection ratio of TN was 67.8% ± 7.3%. In general, TN rejection ratios in

most studies changed between 50%-80%. With regard to the ABM, Liu et al. [39] developed

an experiment in a microbial fuel cell combined with ABM FO membrane, and found that the

rejection ratio of TN was only 29.4%, which was still in the scope in Fig. 1(d).

Further experiments were designed to figure out the retention abilities of ammonia and

nitrate using ABM, with 5 mg/L (NH4+-N) ammonia chloride solution and 5 mg/L (NO3--N)

potassium nitrate solution as the feed solution, respectively, as shown in Fig. 2(a). After a

11
running period of 20 h, the rejection ratio of ammonia by the ABM is very low, and most

ammonia was transferred from the feed side to the draw side. On the contrary, the average

rejection ratio of nitrate reached up to 85.0 %. The results were consistent with Gu et al. [29]

reporting that ammonia was the main form of nitrogen permeating through the CTA FO

membrane, while other forms of nitrogen could be well rejected. However, there are also

some inconsistent results of high rejection ratios of NH4+-N when CTA membranes [11,40] or

TFC membranes [12] was used. The study by Liu et al. [41] even shows that CTA FO

membrane has better rejection for ammonium than for TOC (total organic carbon) and TN. On

the whole, the inconsistent results illustrated that the nitrogen retention ability of FO

membrane should be affected by many factors, which deserve further research in the future.

(Fig. 2)

It's interesting that after 20 h filtration, the NH4+-N concentration in the draw solution

was significantly higher than that in the feed solution (Fig. 2(a)). This phenomenon could be

well explained by the cation exchange theory raised by Lu et al. [42]. With TFC membrane,

when inorganic electrolyte solution was employed as draw solution, more cations in the

solution were transferred to the feed side than anions [18], due to the negatively charged

membrane properties of the TFC membrane [43]. Thus NH4+-N in the feed solution was

exchanged from the feed side to the draw side to keep the draw solution neutral, which was

the main impetus of the NH4+-N penetration. It is noteworthy that compared to TFC, ABM

possesses higher negative Zeta potential [15]. Therefore, the significant higher concentrations

12
of NH4+-N in the draw solution after 20 h concentrating process demonstrated the existence of

the cation exchange phenomenon.

The relationship between the NH4+-N/TN ratio and the TN rejection ratio was shown in

Fig. 2(b). The data were collected during the whole experimental period. It was obvious that

TN rejection ratio decrease with the increase of NH4+-N/TN, which was consistent with the

result in Fig. 2(a), and also with the long error bar of TN rejection in Fig. 1(d). The results

indicated that, to improve the TN rejection of the black odorous water by ABM with NaCl

solution as the draw solution, a simple method is to transfer the ammonia to the nitrate before

the FO process. This could be achieved by pre-aeration of the black odorous water, or by a

combination of the biodegradation and the FO, e. g., the osmotic membrane bioreactor

(OMBR).

Much fluctuation of TN rejection was observed at high NH4+-N/TN value, as shown by

the arrows in Fig. 2(b). This may be due to the different NH4+-N/NO2--N conversion rates by

AOB in the biofouling layer on the membrane surface. During the whole experimental

process, AOB communities in the biofouling layer were regularly detected. The α-diversity,

including the community richness and the community diversity, could reflect the coexistence

of species within a community competing for resources or using the same habitat [44]. Both

the richness (indicated by the index ace/chao) and the diversity (indicated by the index

shannon/simpson) of AOB in the biofouling layer decreased over running cycles, shown in

Table S2 in the supplementary materials. Totally 13 AOB genera were detected, and the

number of genera also decreased (Fig. S3; Fig. S4). The Venn diagram of the AOB at genus

level (Fig. S3) shows that all the detected AOB genera in the fouling layer originated from the

13
influent (M0). At the end of the experiments, only two genera (Nitrosomonas,

norank_o_environmental samples_c_Betaproteobacteria) in the heatmap (Fig. S4) were

abundant in sample M4, whose sequencing numbers were above 4900. Since the data about

TN rejection ratio in Fig. 2(b) were collected at different stages, the differences of AOB

richness and diversity in the biofouling layer would bring about different NH4+-N/NO2--N

conversion rates, which would consequently lead to dramatic fluctuation of the rejection ratio

of TN.

3.2. Membrane fouling characteristics

3.2.1 Water flux decline

The dynamic changes of water flux in the 5 cycles at the interval of uniform time were

shown in Fig. 3(a). The average initial water flux of the five cycles was 9.84 L/(m2·h), higher

than 6-7 L/(m2·h) (ABM, 2 M draw solution) reported by Singh et al. [15] and 5.62 L/(m2·h)

(ABM, 0.5 M draw solution) reported by Liu et al. [39]. The water flux presented a decrease

trend over time in each cycle. It could be explained by the narrowed gap of osmotic pressure

between the draw side and the feed side, where osmotic pressure of feed solution increased

due to concentration while osmotic pressure of draw solution decreased due to the dilution by

the penetrated water [37]. The dots of each cycle overlap well, which meant that there were

no significant differences among the fluxes of the 5 cycles. The average flux of each cycle

was also calculated and shown in Fig. 3(b). Except for some degree of fluctuation in the early

stage of experiment, the average flux in the later period was very stable.

14
(Fig. 3)

It was noteworthy that during the continuous operation of the 64 cycles, 30 min physical

cleaning with deionized water at the end of each cycle was only employed as the membrane

recovery strategy. Generally, substantial efforts have been conducted to minimize the

detrimental effects of biofouling, including membrane surface modification, rinsing or

flushing, air-sparging, osmotic backwashing and chemical cleaning [21]. For example, for a

molasses distillery wastewater concentrating with ABM, besides the physical cleaning,

chemical cleaning with NaOH solution was also needed for every three cycles to keep the flux

constant [15]. The stable average flux in Fig. 3(b) indicated that the simple physical cleaning

was suitable for black odorous water concentrating with ABM.

3.2.2 Surface morphology analysis

The SEM and EDS images of the membrane at different stages were shown in Fig. 4. The

SEM image of the active layer of the new membrane (Fig. 4(a2)) showed that protein vesicles

were densely sprinkled on the surface, similar to the results of Singh et al. [15]. A biofouling

layer gradually formed and thickened with the proceeding of the experiments under current

membrane recovery strategy with physical cleaning, which could be clearly revealed by

comparing Fig. 4 (b1) and (c1). However, the microstructures of the biofouling layers of the

32nd cycle (Fig. 4 (b2)) and the 64th cycle (Fig. 4 (c2)) were similar, presenting a large

number of cocci embedded on the biofilm. The EDS images showed the dynamic changes of

the most abundantly presented elements on the active layer during the whole experiments.

15
The relative weight percentages were also shown in Table S3, presenting a decrease of the

element C combined with obvious increases of the elements P, Mn, Fe and Na in the

biofouling layer.

(Fig. 4)

The distribution of the proteins (stained by FITC) and α-d-glucopyranose polysaccharides

(Stained by Con A) in the membrane fouling layer was further observed by CLSM. The

scanned regime was 1.4 ×1.4 mm2 in aera, and 120 μm in depth. The results were shown in

Fig. 5. Polysaccharides and proteins are two main components of the extracellular polymeric

substances (EPS) of biological treating system [45,46], the distribution of which could well

instruct the biofouling layer characteristics. Fig. 5 shows that, compared to the biofouling

layer on the 32nd day, the red light representing the α-d-glucopyranose polysaccharides

increased (Fig. 5(b2)) while the green light representing the proteins decreased (Fig. 5(b3)),

and the total light intensity increased (Fig. 5(b2)). The results were consistent with those in

the thicker fouling layer in Fig. 4(c1). The higher fraction of polysaccharides as compared to

proteins in membrane fouling layer has also been found in other FO systems [39].

(Fig. 5)

The increases of the inorganic elements P, Mn, Fe and Na shown in Table S3 could be

explained by the characteristics of forward osmosis process. As shown in Fig. 1, TP, the

16
essential element needed for the metabolism of bacterial in the biofouling layer, could be

effectively rejected by the ABM. Therefore, it was not difficult to understand its accumulation

in the fouling layer. The increasing weight percentage of Na was due to the inevitable reverse

salt flux [18]. Fe and Mn are two typical major blackening metals in black-odorous urban

rivers which could form black chelates with humic substances in the water [33]. These

chelates as macromolecular organics could be effectively retained by the FO membrane and

thus accumulated in the biofouling layer [13]. It should be emphasized that the accumulation

of these inorganic elements did not lead to the water flux decline during the whole process

(Fig. 3).

Generally, biofouling will develop over time on the membrane surface regardless of the

pretreatment technologies [21]. The fouling layer of FO membrane was reported to be helpful

for improving pollutant retention ability of the membrane, especially for hydrophobic

compounds [40]. With regard to the water flux, FO biofouling, however, has a significantly

lower impact on permeate water flux than the biofouling in pressurized membrane processes

[47]. A numerical model raised by Bucs et al. [48] even demonstrated that a highly permeable

biofilm could reach a thickness as high as above 100 μm without affecting permeated water

flux. The results of this study showed that, although a simple physical cleaning was employed,

both high pollutant rejection ratio and constant water flux could be achieved when employing

the ABM membrane for black odorous water concentrating.

3.3. Microbial population structure in the bio-fouling layer

Table 1 shows the estimators denoting the α-diversity of the microbial communities of the

biofouling layer at different running stages. The sample M0 referred to the community status

17
of the eutrophic pond. The coverage of each samples was higher than 0.998, illustrating

adequate sequencing depth. It could be found that among the different experimental stages,

the biofouling layer of the 16th cycle (M1) had the lowest bacterial richness (as shown by the

indexes "ace" and "chao"), while the highest richness occurred in the 64th cycle (M4), which

was consistent with that of the progressive thickening biofouling layer (Fig .4, Fig. 5). There

was no obvious difference among the diversities of the samples (as denoted by the indexes

"shannon" and "simpson"), fluctuating around the sample M0. The Venn diagram among

samples M0, M1 and M4 at phylum level (Fig. S5) also showed that totally 14 phyla were

shared by the three samples, and the numbers of the specific phyla in sample M0, M1and M4

were only 4, 1 and 2, respectively, which was consistent with the indexes in Table 1.

Table 1. Bacterial richness and diversity of the 5 samples (Cutoff = 0.03).

Sample ace chao shannon simpson coverage

M0 447 447 3.88 0.0485 0.998647
M1 476 486 3.20 0.1189 0.998253
M2 520 525 4.11 0.0470 0.998577
M3 485 498 3.96 0.0463 0.997804
M4 594 586 3.60 0.0780 0.997517

The relative abundances of taxon assignments at phylum and genus levels and the

function prediction about the bacterial communities of the samples are shown in Fig. 6.

(Fig. 6)

At phylum level, totally 27 phyla were observed, and the 9 most abundant phyla were

shown in Fig. 6 (a). There were similar community structures among the samples M1, M2,
18
M3 and M4 at phylum level, consistent with the study results of Ji et al. [49]. The most

abundant phyla were Proteobacteria, Planctomycetes, Bacteroidetes, Actinobacteria, and

Cyanobacteria, with relative abundances in the sample M4 of 67.5%; 16.3%, 7.2%, 4.6% and

2.5%, respectively. Proteobacteria was also the most dominant phylum (63.2%) in the

influent (M0), the abundance of which was positively correlated with TP and COD [50]. It

was reported to be the largest phylum in most lake or river sediments [51]. Bacteroidetes are

common aerobic bacteria which could promote the degradation of organic matter [3].

Actinobacteria was also found to be abundant in wastewater treatment system [52], drinking

water biofilms [53] and seawater [54], contributing to carbon substrate use. The abundance of

Cyanobacteria, the most widely distributed group of photosynthetic prokaryotes, is due to its

ability of rapid proliferation in eutrophic water [55]. It's noteworthy that the phylum

Planctomycetes only accounted for 0.4% in M0, while its relative abundance increased

sharply in the biofouling layer. The phylum was widely found in aquatic environment [56],

mainly characterized by an attached lifestyle and mostly dominates biofilms [57].

At genus level, there are 420 genera detected totally, in which 63 genera were specific in

M0. Fig. 6(b) exhibits the 26 most abundant genera in the five samples. The dominant genera

with relative percentages above 2.0% in samples M0 and/or M4 were shown in Table 2. Three

genera were abundant in both samples, accounting for 9.4% in M4 and 22.8% in M0. Five

genera accounted for 56.8% in M4, while their total accounting percentage was only 0.041%

in M0. On the other hand, 8 genera dominant in M0 reached a total percentage of 47.6%,

while their total content in M4 only accounted for 1.6%. The results indicated that the

dominant bacterial community in the fouling layer was quite different from that in the black

19
odorous water. The genera norank_f__Reyranellaceae boosted rapidly from 0.0027% in M0

to 23.5% in M4. This genus belongs to the class Alphaproteobacteria, the dominance of

which in M4 may be related to the concentrating organic and nutrient pollutants of the feed

solution [50]. Erythrobacter is reported to be one of the most abundant groups in eutrophic

coastal environments, capable of degrading refractory carbon [58]. The enrichment of this

genus in M4 was due to both substrates concentrating and sodium accumulation reversely

penetrating from the draw side (Table S3). SM1A02 was thought to be associated with

nitrogen removal, possessing the function of anammox [59]. Pirellula was also a typical

anaerobic ammonium oxidizing bacteria strongly adapted to the saline environment, and its

predominant in M4 may be also related to the sodium accumulation in the biofouling layer

[60]. Sphingopyxis was reported to have the capacity of degrading petroleum hydrocarbons

[61]. Hydrogenophaga are facultative autotrophs contributing to hydrogenotrophic

denitrification process [62]. They are abundant in samples M0 and M4, simultaneously. In

general, compared to sample M0, the boost of these genera in M4 may be due to either

concentrating pollutants in the feed solution or salt accumulation in the biofouling layer.

These genera would lead to a complex pollutant transformation (especially nitrogen

transformation) during the FO process, which still need further studies.

Community structures of the four membrane fouling layer samples were also compared.

The Venn diagram of the bacterial communities at genus level among samples M1, M2, M3

and M4 was shown in Fig. S6. Up to 167 genera were shared by the 4 samples. Only 7, 19, 9

and 24 genera were specific in M1, M2, M3 and M4, respectively The top 10 abundant genera

in the four membrane fouling layer samples were listed in Table S4. Among these dominant

20
 genera, 2 genera (Hydrogenophaga and Gordonia) were detected in all samples, 4 genera

(unclassified_f__Burkholderiaceae, norank_f__Reyranellaceae, Erythrobacter and

Sphingopyxis) appeared in at least 3 samples, and 3 genera (Marinobacter, Pirellula and

norank_o__Chloroplast) in at least 2 samples. These results indicated that the bacterial

community structures of the biofouling layer at different stages were similar, which was

consistent with the bacterial diversity in Table 1.

Table 2. The most abundant genera with relative percentages above 2.0% in sample M0 and/or

M4.

Percentage (%)
Phylum Genus
M4 M0

Abundant Proteobacteria Hydrogenophaga 5.0 6.2

in both Proteobacteria unclassified_f__Burkholderiaceae 2.2 13.1
samples Cyanobacteria norank_o__Chloroplast 2.2 3.5
Proteobacteria norank_f__Reyranellaceae 23.5 0.0027

Proteobacteria Erythrobacter 17.1 0

Abundant
Planctomycetes SM1A02 9.9 0
only in M4
Planctomycetes Pirellula 4.2 0.038
Proteobacteria Sphingopyxis 2.1 0
Proteobacteria C39 0.62 16.4
Actinobacteria hgcI_clade 0.17 5.3

Proteobacteria norank_f__Hydrogenophilaceae 0.023 5.2

Abundant Actinobacteria norank_f__Sporichthyaceae 0.033 5.2

only in M0 Proteobacteria Polynucleobacter 0.18 4.9
Actinobacteria unclassified_f__Sporichthyaceae 0.095 3.9
Proteobacteria norank_f__Methylophilaceae 0.074 3.7
Bacteroidetes norank_o__OPB56 0.38 3.0

21
 PICRUSt functional predictive analysis about the 4 biofouling layer samples was further

conducted based on the sequencing data (Fig. 6(c)). The functional annotation of COG and

KEGG of OTU was carried out by the greengene ID corresponding to each OTU, which has

been standardized by PICRUSt software package. It could be found that the bacterial

communities in the membrane fouling layer were associated with 25 functional gene families.

The abundance information of each function in different samples was very similar, which was

consistent with the Venn diagram (Fig. S6). Detailed abundance values of genes assigned in

different functional categories of sample M4 were shown in Fig. S7. The top 6 most dominant

COG functional groups were Amino acid transport and metabolism, General function

prediction only, Energy production and conversion, Cell wall/membrane/envelope biogenesis,

Signal transduction mechanisms, and Inorganic ion transport and metabolism. These dominant

predicted functions were similar with those of the bacterial communities in a municipal

wastewater treatment plant [59].

4. Conclusions

ABM was employed for black odorous water concentrating in this study. Most pollutants

in the black odorous water, including organic matters (COD), TP and nitrate, could be

effectively rejected by the FO membrane. Due to cation exchange, NH4+-N penetrated from

the feed side to the draw side when NaCl was employed as the draw solution. The TN

rejection characteristic, to a large extent, depended on the NH4+-N/TN ratio. TN rejection

could be evidently improved by transferring ammonia in the black odorous water to nitrate

before the FO process. Succession of the AOB communities enriched in the biofouling layer

22
of the membrane surface would also affect the transformation of NH4+-N and thus lead to

fluctuations of TN rejection ratio especially at high NH4+-N/TN ratio. With a physical

cleaning at the end of each cycle, the FO process could achieve a constant water flux during

the 64-cycle continuous operation. CLSM showed that the concentration of polysaccharides in

the biofouling layer increased while the concentration of proteins decreased at the end of the

experiments. The accumulation of the elements P, Mn, Fe and Na was also observed in the

fouling layer. Bacterial richness of the biofouling layer increased over the running period. The

most dominant genera in M4 included norank_f__Reyranellaceae, Erythrobacter, SM1A02,

Pirellula, Hydrogenophaga, etc. The boost of these genera in the biofouling layer would lead

to a complex pollutant transformation (especially nitrogen transformation) during the FO

process.

23
Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal

relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

This work was financially supported by the Open Research Fund of Jiangsu Province

Key Laboratory of Environmental Engineering (KF2018003), the Natural Science Foundation

of Jiangsu Province of China (No. BK20171478), and the Priority Academic Program

Development of Jiangsu Higher Education Institutions (No. 164320H116).

24
References

[1] H. Yin, P. Yang, M. Kong, Effects of nitrate dosing on the migration of reduced sulfur in

black odorous river sediment and the influencing factors, Chem. Eng. J. 371 (2019)

516-523.

[2] M. Hamzeh, B. Ouddane, M. Daye, J. Halwani, Trace metal mobilization from surficial

sediments of the seine river estuary, Water Air Soil Pollut. 225 (2014) 1878.

[3] W.H. Wang, Y. Wang, P. Fan, L.F. Chen, B.H. Chai, J.C. Zhao, L.Q. Sun, Effect of

calcium peroxide on the water quality and bacterium community of sediment in

black-odor water, Environ. Pollut. 248 (2019) 18-27.

[4] M. Pan, J. Zhao, S. Zhen, S. Heng, J. Wu, Effects of the combination of aeration and

biofilm technology on transformation of nitrogen in black-odor river, Water Sci. Technol.

74 (2016) 655-662.

[5] D. Copetti, K. Finsterle, L. Marziali, F. Stefani, G. Tartari, G. Douglas, K. Reitzel, B.M.

Spears, I.J. Winfield, G. Crosa, Eutrophication management in surface waters using

lanthanum modified bentonite: a review, Water Res. 97 (2016) 162-174.

[6] Z. Zhou, T. Huang, B. Yuan, Nitrogen reduction using bioreactive thin-layer capping

(BTC) with biozeolite: a field experiment in a eutrophic river, J. Environ. Sci. 42 (2016)

119-125.

[7] C. Liu, Q. Shen, Q. Zhou, C. Fan, S. Shao, Precontrol of algae-induced black blooms

through sediment dredging at appropriate depth in a typical eutrophic shallow lake, Ecol.

Eng. 77 (2015) 139-145.

25
[8] B. Pan, J. Yuan, X. Zhang, Z. Wang, J. Chen, J. Lu, W. Yang, Z. Li, N. Zhao, M. Xu, A

review of ecological restoration techniques in fluvial rivers, Int. J. Sediment Res. 31

(2016) 110-119.

[9] S. Kim, K.H. Chu, Y.A.J. Al-Hamadani, C.M. Park, M. Jang, D.H. Kim, M. Yu, J. Heo,

Y. Yoon, Removal of contaminants of emerging concern by membranes in water and

wastewater: A review, Chem. Eng. J. 335 (2018) 896-914.

[10] A.J. Ansari, F.I. Hai, W.E. Price, J.E. Drewes, L.D. Nghiem, Forward osmosis as a

platform for resource recovery from municipal wastewater - A critical assessment of the

literature, J. Membr. Sci. 529 (2017) 195-206.

[11] L. Chen, Y.S. Gu, C.Q. Cao, J. Zhang, J.W. Ng, C.Y. Tang, Performance of a submerged

anaerobic membrane bioreactor with forward osmosis membrane for low-strength

wastewater treatment, Water Res. 50 (2014) 114-123.

[12] W. Luo, H.V. Phan, M. Xie, F.I. Hai, W.E. Price, M. Elimelech, L.D. Nghiem, Osmotic

versus conventional membrane bioreactors integrated with reverse osmosis for water

reuse: biological stability, membrane fouling, and contaminant removal, Water Res. 109

(2017) 122-134.

[13] L. Chekli, S. Phuntsho, J.E. Kim, J. Kim, J.Y. Choi, J.S. Choi, S. Kim, J.H. Kim, S. Hong,

J. Sohn, H.K. Shon, A comprehensive review of hybrid forward osmosis systems:

Performance, applications and future prospects, J. Membr. Sci. 497 (2016) 430-449.

[14] Z. Li, R.V. Linares, S. Bucs, L. Fortunato, C. Hélix-Nielsen, J.S. Vrouwenvelder, N.

Ghaffour, T. Leiknes, G. Amy, Aquaporin based biomimetic membrane in forward

osmosis: Chemical cleaning resistance and practical operation, Desalination 420 (2017)

26
 208-215.

[15] N. Singh, I. Petrinic, C. Hélix-Nielsen, S. Basu, M. Balakrishnan, Concentrating

molasses distillery wastewater using biomimetic forward osmosis (FO) membranes,

Water Res. 130 (2018) 271-280.

[16] X. Wang, T. Hu, Z. Wang, X. Li, Y. Ren, Permeability recovery of fouled forward

osmosis membranes by chemical cleaning during a long-term operation of anaerobic

osmotic membrane bioreactors treating low-strength wastewater, Water Res. 123 (2017)

505-512.

[17] T. Xiao, P. Dou, J. Wang, J. Song, Y. Wang, X.M. Li, T. He, Concentrating greywater

using hollow fiber thin film composite forward osmosis membranes: Fouling and process

optimization, Chem. Eng. Sci. 190 (2018) 140-148.

[18] W. Cheng, X. Lu, Y. Yang, J. Jiang, J. Ma, Influence of composition and concentration of

saline water on cation exchange behavior in forward osmosis desalination, Water Res.

137 (2018) 9-17.

[19] F. Volpin, E. Fons, L. Chekli, J.E. Kim, A. Jang, H.K. Shon, Hybrid forward

osmosis-reverse osmosis for wastewater reuse and seawater desalination: Understanding

the optimal feed solution to minimise fouling, Process Saf. Environ. 117 (2018) 523-532.

[20] Y. Sun, J. Tian, L. Song, S. Gao, W. Shi, F. Cui, Dynamic changes of the fouling layer in

forward osmosis based membrane processes for municipal wastewater treatment, J.

Membr. Sci. 549 (2018) 523-532.

[21] A. Bogler, S. Lin, E. Bar-Zeev, Biofouling of membrane distillation, forward osmosis and

pressure retarded osmosis: Principles, impacts and future directions, J. Membr. Sci. 542

27
 (2017) 378-398.

[22] S. You, J. Lu, C.Y. Tang, X. Wang, Rejection of heavy metals in acidic wastewater by a

novel thin-film inorganic forward osmosis membrane, Chem. Eng. J. 320 (2017)

532-538.

[23] SEPA, Water and Wastewater Monitoring Methods, fourth ed., Chinese Environ-mental

Science Publishing Press, Beijing, China, 2002.

[24] M.Y. Chen, D.J. Lee, Z. Yang, X.F. Peng, J.Y. Lai, Fluorecent staining for study of

extracellular polymetic substances in membrane biofouling layers, Environ. Sci. Technol.

40 (2006) 6642-6646.

[25] Z.J. Wang, Z. Wang, K. Xu, L. Chen, Z.Z. Lin, Y.L. Liu, Performance evaluation of

adsorptive removal of Ni(II) by treated waste granular-activated carbon and new

granular-activated carbon, Desalin. Water Treat. 161 (2019) 315-326.

[26] M.Z.M. Yusoff, A. Hu, C. Feng, T. Maeda, Y. Shirai, M.A. Hassan, C.P. Yu, Influence of

pretreated activated sludge for electricity generation in microbial fuel cell application,

Bioresour. Technol. 145 (2013) 90-96.

[27] Y. Mao, A.C. Yannarell, R.I. Mackie, Changes in N-transforming archaea and bacteria in

soil during the establishment of bioenergy crops, PLOS One 6 (2011) e24750.

[28] K.R. Amato, C.J. Yeoman, A. Kent, N. Righini, F. Carbonero, A. Estrada, H.R. Gaskins,

R.M. Stumpf, S. Yildirim, M. Torralba, M. Gillis, B.A. Wilson, K.E. Nelson, B.A. White,

S.R. Leigh, Habitat degradation impacts black howler monkey (Alouatta pigra)

gastrointestinal microbiomes, ISME J. 7 (2013) 1344-1353.

[29] Y. Gu, L. Chen, J.W. Ng, C. Lee, V.W.C. Chang, C.Y. Tang, Development of anaerobic

28
 osmotic membrane bioreactor for low-strength wastewater treatment at mesophilic

condition, J. Membr. Sci. 490 (2015) 197-208.

[30] N.D. Viet, J. Cho, Y. Yoon, A. Jang, Enhancing the removal efficiency of osmotic

membrane bioreactors: A comprehensive review of influencing parameters and hybrid

configurations, Chemosphere 236 (2019) 1-18.

[31] H.T. Madsen, N. Bajraktari, C. Hélix-Nielsen, B. van der Bruggen, E.G. Søgaard, Use of

biomimetic forward osmosis membrane for trace organics removal, J. Membr. Sci. 476

(2015) 469-474.

[32] M. Xu, Q. Zhang, C. Xia, Y. Zhong, G. Sun, J. Guo, T. Yuan, J. Zhou, Z. He, Elevated

nitrate enriches microbial functional genes for potential bioremediation of complexly

contaminated sediments, ISME J. 8 (2014) 1932-1944.

[33] Z. Liang, M. Siegert, W. Fang, Y. Sun, F. Jiang, H. Lu, G.H. Chen, S. Wang, Blackening

and odorization of urban rivers: a bio-geochemical process, FEMS Microbiol. Ecol. 94

(2018) fix180.

[34] T. Melin, B. Jefferson, D. Bixio, C. Thoeye, W. de Wilde, J. de Koning, T. Wintgens,

Membrane bioreactor technology for wastewater treatment and reuse, Desalination 187

(2006) 271-282.

[35] Z. Wu, S. Zou, B. Zhang, L. Wang, Z. He, Forward osmosis promoted in-situ formation

of struvite with simultaneous water recovery from digested swine wastewater, Chem.

Eng. J. 342 (2018) 274-280.

[36] S. Zou, Z. He, Enhancing wastewater reuse by forward osmosis with self-diluted

commercial fertilizers as draw solutes, Water Res. 99 (2016) 235-243.

29
[37] R.V. Linares, Z. Li, M. Abu-Ghdaib, C.H. Wei, G. Amy, J.S. Vrouwenvelder, Water

harvesting from municipal wastewater via osmotic gradient: an evaluation of process

performance, J. Membr. Sci. 447 (2013) 50-56.

[38] Z. Wang, J. Zheng, J. Tang, X. Wang, Z. Wu, A pilot-scale forward osmosis membrane

system for concentrating low-strength municipal wastewater: performance and

implications, Sci. Rep. 6 (2016) 21653.

[39] J. Liu, X. Wang, Z. Wang, Y. Lu, X. Li, Y. Ren, Integrating microbial fuel cells with

anaerobic acidification and forward osmosis membrane for enhancing bio-electricity and

water recovery from low-strength wastewater, Water Res. 110 (2017) 74-82.

[40] N.C. Nguyen, S.S. Chen, H.Y. Yang, N.T. Hau, Application of forward osmosis on

dewatering of high nutrient sludge, Bioresour. Technol. 132 (2013) 224-229.

[41] Q. Liu, C. Liu, L. Zhao, W. Ma, H. Liu, J. Ma, Integrated forward osmosis-membrane

distillation process for human urine treatment, Water Res. 91 (2016) 45-54.

[42] X. Lu, C. Boo, J. Ma, M. Elimelech, Bidirectional diffusion of ammonium and sodium

cations in forward osmosis: role of membrane active layer surface chemistry and charge,

Environ. Sci. Technol. 48 (2014) 14369-14376.

[43] D.X. Hou, L. Lu, Z.J. Ren, Microbial fuel cells and osmotic membrane bioreactors have

mutual benefits for wastewater treatment and energy production, Water Res. 98 (2016)

183-189.

[44] E. Xiao, Y. Zhou, D. Xu, R. Lu, Y. Chen, Q. Zhou, Z. Wu, The physiological response of

Arundo donax and characteristics of anodic bacterial community in BE-CW systems:

Effects of the applied voltage, Chem. Eng. J. 380 (2020) 122604.

30
[45] H.L. Zhang, W.L. Jiang, R. Liu, Y. Zhou, Y. Zhang, Organic degradation and

extracellular products of pure oxygen aerated activated sludge under different F/M

conditions, Bioresour. Technol. 279 (2019) 189-194.

[46] Y.Q. Hu, W. Wei, M. Gao, Y. Zhou, G.X. Wang, Y. Zhang, Effect of pure oxygen aeration

on extracellular polymeric substances (EPS) of activated sludge treating saline

wastewater, Process Saf. Environ. 123 (2019) 344-350.

[47] S.E. Kwan, E. Bar-Zeev, M. Elimelech, Biofouling in forward osmosis and reverse

osmosis: measurements and mechanisms, J. Membr. Sci. 493 (2015) 703-708.

[48] S.S. Bucs, R.V. Linares, J.S. Vrouwenvelder, C. Picioreanu, Biofouling in forward

osmosis systems: an experimental and numerical study, Water Res. 106 (2016) 86-97.

[49] Y. Ji, R. Angel, M. Klose, P. Claus, H. Marotta, L. Pinho, A. Enrich-Prast, R. Conrad,

Structure and function of methanogenicmicrobial communities in sediments of

Amazonian lakes with different water types, Environ. Microbiol. 18 (2016) 5082-5100.

[50] T. Zhang, M. Shao, L. Ye, 454 Pyrosequencing reveals bacterial diversity of activated

sludge from 14 sewage treatment plants, ISME J. 6 (2012) 1137-1147.

[51] D. Savio, L. Sinclair, U.Z. Ijaz, J. Parajka, G.H. Reischer, P. Stadler, A.P. Blaschke, G.

Blöschl, R.L. Mach, A.K.T. Kirschner, A.H. Farnleitner, A. Eiler, Bacterial diversity

along a 2600 km river continuum, Environ. Microbiol. 17 (2015) 4994-5007.

[52] L. Zhang, G. Fu, Z. Zhang, Simultaneous nutrient and carbon removal and electricity

generation in selfbuffered biocathode microbial fuel cell for high-salinity mustard tuber

wastewater treatment, Bioresour. Technol. 272 (2019) 105-113.

[53] Y. Chao, Y. Mao, Z. Wang, T. Zhang, Diversity and functions of bacterial community in

31
 drinking water biofilms revealed by high-throughput sequencing, Sci. Rep. 5 (2015)

10044.

[54] A. Belila, J. El-Chakhtoura, N. Otaibi, G. Muyzer, G. Gonzalez-Gil, P.E. Saikaly, M.C.M.

van Loosdrecht, J.S. Vrouwenvelder, Bacterial community structure and variation in a

full-scale seawater desalination plant for drinking water production, Water Res. 94 (2016)

62-72.

[55] L.J. Wang, G. Pan, W.Q. Shi, Z.B. Wang, H.G. Zhang, Manipulating nutrient limitation

using modified local soils: a case study at Lake Taihu (China), Water Res. 101 (2016)

25-35.

[56] X.C. Ma, X.K. Li, X.W. Wang, G.G. Liu, J.L. Zuo, S.T. Wang, K. Wang, Impact of

salinity on anaerobic microbial community structure in high organic loading purified

terephthalic acid wastewater treatment system, J. Hazard. Mater. 383 (2020) 121132.

[57] M.M. Bengtsson, L. Øvreås, Planctomycetes dominate biofilms on surfaces of the

Kelplaminaria hyperborea, BMC Microbiol. 10 (2010) 261-270.

[58] S. Vanucci, F. Guidi, R. Pistocchi, R.A. Long, Phylogenetic structure of bacterial

assemblages co-occurring with Ostreopsis cf. ovata bloom, Harmful Algae 55 (2016)

259-271.

[59] L. Zhang, Z. Shen, W. Fang, G. Gao, Composition of bacterial communities in municipal

wastewater treatment plant, Sci. Total Environ. 689 (2019) 1181-1191.

[60] Z. Xia, Q. Wang, Z. She, M. Gao, Y. Zhao, L. Guo, C. Jin, Nitrogen removal pathway

and dynamics of microbial community with the increase of salinity in simultaneous

nitrification and denitrification process, Sci. Total Environ. 697 (2019) 134047.

32
[61] A.L. Angelim, S.P. Costa, B.C.S. Farias, L.F. Aquino, V.M.M. Melo, An innovative

bioremediation strategy using a bacterial consortium entrapped in chitosan beads, J.

Environ. Manage. 127 (2013) 10-17.

[62] W. Xing, J. Li, P. Li, C. Wang, Y. Cao, D. Li, Y. Yang, J. Zhou, J. Zuo, Effects of residual

organics in municipal wastewater on hydrogenotrophic denitrifying microbial

communities, J. Environ. Sci. 65 (2018) 262-270.

33
Figure captions

Fig. 1. Concentrating effects of COD (a), TN (b), TP (c), and the rejection ratios of the three

pollutants in the black odorous water (d).

Fig. 2. Nitrogen Concentrating characteristics: (a) Concentrations of NH4+, NO3- in the feed

solution and the draw solution after FO filtration. The initial feed solutions were NH4Cl

solution (with the NH4+-N concentration of 5 mg/L) and KNO3 solution (with the NO3--N

concentration of 5 mg/L), respectively. (b) The relationship between the rejection ratios of TN

and the percentage of NH4+-N in TN of the black odorous water.

Fig. 3. (a) Dynamic changes of water flux in the 1st cycle, the 16th cycle, the 32nd cycle and

the 64th cycle, (b) the average water flux over the 64 cycles duration of the experiment.

Fig. 4. SEM images of the active layer of the new membrane surface (a) and the pollutant

layers of the 32nd cycle membrane surface (b) and the 64th cycle membrane surface (c), in

which a1, b1 and c1 were observed at 100×, a2, b2 and c2 were observed at 5000×, a3, b3

and c3 were the EDS images of the new membrane, the pollutant layer of the 32nd cycle

membrane surface and the 64th cycle membrane surface, respectively.

Fig. 5. CLSM images of the fouling layers of the 32nd membrane surface (a1-a4) and the 64th

membrane surface (b1-b4), respectively. (a1) and (b1): combined 3D images of

α-polysaccharides and proteins, (a2) and (b2): section images of α-polysaccharides (Con A,

red), (a3) and (b3): section images of proteins (FITC, green), (a4) and (b4): combined section

images of α-polysaccharides and proteins.

Fig. 6. Bacterial communities of the 5 samples at the phylum level (a) and genus level (b),

respectively, and the function prediction about the bacterial communities in the fouling layer

of the FO membrane by PICRUSt software package (c).

34
35
36
Declaration of interests

37
☒ The authors declare that they have no known competing financial interests or personal relationships
that could have appeared to influence the work reported in this paper.

☐The authors declare the following financial interests/personal relationships which may be
considered as potential competing interests:

Highlights:

 Black odorous water was concentrated by aquaporin biomimetic membrane for further

treatment.

 TN rejection ratio was largely dependent upon the NH4+-N/TN ratio.

 AOB community affected NH4+-N transformation and caused TN rejection fluctuation.

 The elements P, Na and the blackening metals Fe and Mn accumulated in the fouling

layer.

 Water flux decline was not observed with physical cleaning only.

38