Chemosphere 307 (2022) 135654

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Chemosphere
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Biodegradation and optimization of bilge water in a sequencing batch

reactor using response surface methodology
Jianqiang Shi a, *, Guichen Zhang a, Shaojun Zhang b, Run Lu a, Mengwei Chen a
a
College of Merchant Marine, Shanghai Maritime University, Shanghai, 201306, China
b
School of Navigation and Shipping, Shandong Jiaotong University, Weihai, 264200, China

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• A sequencing batch reactor is proposed

to treat bilge water.
• The operating parameters were opti­
mized using the response surface
method.
• The practical performance of
sequencing batch reactor is explored.
• The first-order biodegradation kinetic
model is obtained under the optimal
conditions.

A R T I C L E I N F O A B S T R A C T

Handling Editor: Pau-Loke SHOW Bilge water is a significant source of pollution in the marine environment and has captured widespread inter­
national attention. In this study, a sequencing batch reactor (SBR) combined with strain S2 identified as Bacillus
Keywords: licheniformis was employed to assess the biodegradation of Chemical Oxygen Demand (COD) from bilge water.
Bilge water The influencing variables such as temperature, pH level and inoculum concentration on the performance SBR
COD
system were optimized by utilizing response surface methodology (RSM). The experimental results showed that
Biodegradation
the maximum COD removal of 77.81% was reached at the optimal SBR operation conditions of temperature
RSM
Kinetic 35.44 ◦ C pH 8.13, and inoculum concentration 31.47 mL. In the practical application of SBR, it was found that
the decrease in hydraulic retention time (HRT) was accompanied by a decrease in COD degradation rate. The
biodegradation kinetics of COD in bilge water were well fitted with the first–order equation with a higher R2
value of 0.98106. In conclusion, COD in bilge water can be efficiently biodegraded by SBR under the optimi­
zation of RSM.

1. Introduction
The oily wastewater mainly originated from leakages of pipes and
pumps or the condensation of vapor is called bilge water, which is not
only a mixture of the residuals from diesel oil, grease, and cleaning agent
but also contains seawater, organic compounds, suspended solids, and
heavy metals (Uma and Gandhimathi, 2019; Gryta, 2020). Moreover,
the production and composition of a ship’s bilge water differ signifi­
cantly due to the types and operation management of each ship (Mag­
nusson et al., 2018), but as a rule, all bilge water contaminants are

* Corresponding author.
E-mail address: stoneseaman1228@163.com (J. Shi).

https://doi.org/10.1016/j.chemosphere.2022.135654
Received 29 April 2022; Received in revised form 23 June 2022; Accepted 5 July 2022
Available online 18 July 2022
0045-6535/© 2022 Elsevier Ltd. All rights reserved.
J. Shi et al.
detrimental to the natural ocean environment and simultaneously
constitute enormous potential harm to the marine biodiversity (Gatidou
et al., 2022).
The International Maritime Origination (IMO) had mandatorily
regulated the bilge water discharge standard (less than 15 ppm of oil
concentration). Effective and feasible treatment technology adapted for
bilge water treatment is enforced for both shipboard and pier-side to
meet the relevant requirements. At present, a conventional oily water
separator was widely equipped on-board, which is incapable of purify­
ing the bilge water with the oil droplets smaller than 20 μm since the oil-
in-water emulsions formed by the surfactants derived from the cleaning
agent are chemically stabilized (Church et al., 2017; Eskandarloo et al.,
2018; Vyrides et al., 2018). In addition, physicochemical methods (e.g.,
membrane filtration, coagulation, electrocoagulation, flotation, floccu­
lation) and other integrated systems utilizing two or three treatment
techniques in the deoiling installation, have been studied as well as
partially applied in previous studies (Lu et al., 2018; Soeprijanto et al.,
2019; Gryta, 2020). However, these processes conduce substantially to
handling costs (Ahmad et al., 2018), and therefore it is imperative to
develop a cost-effective shipboard oil-in-water emulsions treatment
system to satisfy the discharge standards, particularly since an enormous
volume of bilge water increases with the intensive development of
marine transportation (Gryta, 2020).
On the other side, environmentally friendly biological treatment
approaches are available for the degradation of emulsified oil in bilge
water, which could convert organic matters into CO2 and H2O eventu­
ally by the metabolic pathway of hydrocarbon-degrading bacteria (Uma
and Gandhimathi, 2020). Under anaerobic or aerobic conditions,
biodegradation of bilge water has increased enormously in recent years.
Emadian et al. (2015a)and Emadian et al. (2015b) investigated a hybrid
up-flow anaerobic sludge blanket (HUASB) bioreactor and up-flow
anaerobic sludge fixed film (UASFF) reactor, respectively. Conse­
quently, both HUASB and UASFF are preferable for the treatment of
low-strength bilge water. Whereas, the real bilge water wasn’t adopted
in their experiments. Various kinds of biofilm reactors have been
established to remove the oil in other researches (Vyrides et al., 2018;
Mazioti et al., 2021), however, the membrane fouling owing to the
presence of compounds consisting of suspended solids, solvents, and
heavy metals in bilge water, retarded the performance of biological
systems (Li et al., 2021a). The SBR exhibited a preferred application in
organic removal from synthetic oily bilge water utilizing bacterial
consortium (Uma and Gandhimathi, 2019, 2020).
The majority of studies were concerned about the impact of various
SBR operational factors including, cycle time (CT) (Uma and Gan­
dhimathi, 2019), HRT (Santiago et al., 2020; Li et al., 2021b), and solids
retention time (SRT) (Sathian et al., 2014; Santiago et al., 2020) on
soluble COD removal efficiency and SBR optimal operation. Neverthe­
less, the other equally essential factors like temperature, pH level, so­
dium chloride concentration, inoculum concentration, and the
concentration of substrate affecting the biodegradation activities of COD
from bilge water are rarely reported. Moreover, the univariate approach
was widely employed in the optimization of SBR operation, nonetheless,
it cannot reflect the synergistic effects among process parameters (Zhang
et al., 2016). Interestingly, the involvement of RSM can overcome this
trouble. RSM is a statistical technique for the evaluation and optimiza­
tion of conditions, which aims to study the influence of multivariable
parameters on the experimental design and to reduce the cost of
expensive analysis in the meantime (Besharati Fard et al., 2021; Guo
et al., 2021). To the author’s best knowledge, there is no report about the
optimization of operational factors (i.e., temperature, pH level, and
inoculum concentration) for COD removal from bilge water in a SBR.
The purpose of this study was to explore the biodegradation of or­
ganics (in terms of COD) from real bilge water in a discontinuous SBR
using hydrocarbon-degrading bacteria isolated from oil-contaminated
sites. Also, RSM is adopted to analyze and optimize the various pro­
cess conditions (i.e., pH, temperature and inoculum concentration) of
2
Chemosphere 307 (2022) 135654
SBR for the treatment of COD. Furthermore, the practical performance
of SBR at various organic loading rate (OLR) as well as biodegradation
kinetics were also explored under optimized conditions.
2. Materials and methods
2.1. Chemicals and medium
All chemicals like petroleum ether, hydrochloric acid, sodium hy­
droxide, sodium chloride and phosphate buffer were purchased from
SINOPHARM(China) of analytical grade for all experiments. The MSM
chose as the selective medium for bacterial isolation was composed of (g
L− 1) Na2HPO4⋅2H2O 3.5, KH2PO4 1.0, (NH4)2SO4 0.5, MgCl2⋅6H2O 0.1,
Ca(NO3)2⋅4H2O 0.05, EDTA 0.5, FeSO4⋅7H2O 0.2, ZnSO4⋅7H2O 0.1,
MnCl2⋅4H2O 0.03, H3BO3 0.3, CoCl2⋅6H2O 0.2, CuCl2⋅2H2O 0.01,
NiCl2⋅6H2O 0.02, H4MoNa2O6 0.03. The trypticase soy agar (TSA)
containing (g L− 1) tryptone 15.0, soytone 5.0, NaCl 5.0, and agar 13.0,
were prepared for the bacterial enrichment and screen. All mediums
were sterilized in an autoclave at 121 ◦ C for 15 min before use.
2.2. Bilge water characteristic
The bilge water was supplied from a bulk carrier berthing at Longkou
port of China, and the initial COD, pH, and conductive of the obtained
bilge water range from 8100 to 9300 mg L− 1, 7.42 to 7.85, and 11.2 to
11.8 mS cm− 1respectively.
2.3. Inoculum and identification of strain
The acquisition of hydrocarbon-degrading bacteria for SBR was
derived from the oil-contaminated soil sample as per the procedure
designed by (Yang et al., 2018). MSM medium with 1% (v/v) diesel oil
as the sole carbon was employed for the selective media of hydrocarbon
degraders. The obtained bacteria were further subcultured and plated on
TSA. After adequate growth, the plates were deposited at 4 ◦ C for the
following experimental use.
To identify the isolated bacteria, the 16 S rRNA gene sequence of
obtained bacteria was amplified with the extracted DNA using universal
primer 27F5′ -AGA GTT TGA TCM TGG CTC AG-3′ and 1492R5′ -AAG
GAG GTG WTC CAR CC-3’. After comparing it with the Nucleotide
database of National Center for Biotechnology Information (NCBI) using
the BLAST method (Guo et al., 2021), the phylogenetic tree will be
generated to describe the phylogenetic relationship of obtained bacteria
by using the MEGA 7.0 software (Tatar, 2021).
2.4. Batch SBR experiments and RSM optimization design
A biodegradation assessment of organics was performed in a SRB
with a working volume of 2.5 L. During the batch experiments, 5 d of
SRT and 24 h of CT were preferable according to the results implied by
Uma and Gandhimathi (2019) and the fresh air was supplied by an air
diffuser at the rate of 10 LPH (Sathian et al., 2014). During each 24 h of a
cycle, the raw bilge water was pumped into the reactor in 10 min
(feeding step). Continuously, the aeration lasted for 23 h (reaction step).
SRT was maintained by sludge wasting for 10 min (wasting step).
Sedimentation was then operated for 30 min after ceasing the aeration
(settling step). Subsequently, the supernatant would be decanted within
10 min (withdraw step). After that, the above cycle operation was
continued starting with the feeding step. After reaction step, a 10 mL of
sample was collected from the wasted mixed liquor for the determina­
tion of relevant parameters.
The optimization of variables like temperature (20, 25, 30, 35, and
40 ◦ C), pH (5,6,7,8 and 9), and inoculum concentration (10, 20, 30, 40,
and 50 mL) was carried out in each batch SBR experiments referring to
the above procedures. In our preliminary screening studies using the
univariate approach, it is proved that temperature (30, 35, and 40 ◦ C),
J. Shi et al. Chemosphere 307 (2022) 135654

pH (7, 8, and 9), and inoculum concentration (20, 30, and 40 mL) had a 372.08 and P-value of less than 0.0001 (less than 0.05) implies the
noticeable impact on the COD removal. Accordingly, the optimal ex­ model is significant, which means there is only a 0.01% possibility that a
periments were conducted with these three factors at three levels model F-value this large could occur because of noise.
(Table S1). Based on the Box-Behnken design (BBD) of Design-Expert V The P-value of A, B, C, AC, A2, B2 and C2 less than 0.05 indicates the
8.0.6 software (Pinheiro et al., 2021), 17 experiments were imple­ model terms are highly significant. Conversely, a model term like BC is
mented to acquire the experimental data on COD degradation rate, insignificant due to its P-values being greater than 0.1. Because the
which was of paramount importance for obtaining a second-order lower the P-value is, the more significant the matching coefficient is.
polynomial equation and Analysis of variance (ANOVA) analysis (Guo Thus, both pH and inoculum concentration was verified as the most
et al., 2021). At the optimized operational condition, the biodegradation significant parameters affecting COD removal with low P-values (less
kinetic of COD from bilge water was inferred. than 0.0001), followed by temperature with P-values of 0.0002.
The lack of fit with the significance P-value of 0.9594 and F-value of
0.094 implies the adequacy and fitness of this model (Ghelich et al.,
2.5. Analytical methods
2019). Additionally, the R2 with a high value of 0.9979 (close to 1) in­
dicates a good aptness between experimental and predicted data
The pH value was measured by a pH meter (Aicevoos, China) and
(Davarnejad et al., 2014). Furthermore, Adj. R2 and Coefficient of
also the digital display thermometer (LIHUADA, China) was used to
Variation (C⋅V.) were introduced to evaluate the precision and reliability
display the real-time temperature. Determination of COD concentration
of this model (Ghelich et al., 2019). On the one hand, the Adj. R2 value of
was executed in accordance with the standard procedure (Rice et al.,
0.9952 is not only in reasonable agreement with the Pred. R2 value of
2012). In this survey, each experiment was operated in triplicates and
0.9948, but also in proper conformity with R2, which denotes that the
the average results were exhibited.
model possesses a high degree of accuracy and reliability (Besharati Fard
et al., 2021). On the other hand, the C.V. with a low value of 0.49%
3. Results and discussion
states the higher reliability of the experimental data (Davarnejad et al.,
2014). Meanwhile, the Adequate Precision (AP) value of 49.519 greater
3.1. Bacterial identification
than 4 suggests that the fitting model with the adequate signal model
can be used to navigate the design space (Davarnejad et al., 2014; Saber
One bacterial strain named strain S2 with distinct morphological
et al., 2014). Thus, according to the contents above, it can be concluded
features and the ability to biodegrade bilge water was isolated eventu­
that this model is adapted to the theoretical prediction of COD removal
ally. By gene identification, it was found that the strain S2 was matching
and the analysis of interactive effects of variables.
with partial sequence 100% query coverage and 100% identity of Ba­
A modified quadratic model of COD removal in the aspect of the
cillus licheniformis (NCBI Accession Number: NR118996.1) (Fig. S1).
actual factors is expressed by the following equation:
Although, Bacillus licheniformis has been widely investigated in the
bioremediation of all kinds of oil contaminates such as crude oil CODs ​ removal = 73.38 + 0.90A + 1.29B + 1.25C − 0.36AB + 1.04AC
(Kavitha et al., 2014; Liu et al., 2021), heavy oil (Khanpour-Alikelayeh + 0.085BC − 5.76A2 − 4.91B2 − 4.61C2 (1)
et al., 2020), engine oil (Nayak et al., 2020). To date, the application of
Bacillus licheniformis for bilge water disposal was virtually unknown. where, A is the temperature (◦ C), B is pH, and C is inoculum concen­
tration (mL).
3.2. ANOVA and model fitting
3.3. Interactive effects of various parameters on COD removal
Based on the BBD-RSM method, 17 assays were arranged to evaluate
the biodegradation of COD that existed in bilge water and the results The 3D surface curves were plotted to illustrate the interactive effects
were presented in Table S2. The COD removal percentage ranged from of any two independent variables while keeping the rest constant. Fig. 1
65.1% to 78.82% under different factors conditions. As shown in shows the interactive effects of temperature and pH on COD removal
Table 1, the results of ANOVA analysis of the quadratic model for COD while the inoculum concentrate is 30 mL. It can be found that the COD
removal by strain S2 were used to verify the statistical significance and removal increases significantly at most temperatures as the pH increases
reveal the mathematical relationship between independent variables from 7.0 to 9.0, while at most pH values there is only a slight increase in
and COD removal. In this study, the model Fisher’s F value (F-value) of COD removal with increasing temperature from 30 ◦ C to 40 ◦ C, which
reiterates the conclusion obtained in Table 1 regarding the significance
Table 1 of pH over temperature. In addition, COD removal achieves a maximum
ANOVA analysis of the quadratic model. of 78.82% at 35 ◦ C and pH 9, and any deviation from this point would
Source Sum of Degree of Mean F-value P-value result in COD removal reduction. Similarly, Fig. 2 clearly indicates that
squares freedom square the effect of inoculum concentration variation on COD removal is
Model 408.96 9 45.44 372.08 <0.0001 significantly greater than that of temperature while the pH is 8. Because
A-Temperature 6.53 1 6.53 53.20 0.0002 increasing the inoculum concentration from 20 to 40 mL leads to a
B-pH 13.29 1 13.29 108.80 <0.0001 significant increase in COD removal at most temperatures with the
C-Inoculum 12.25 1 12.55 102.76 <0.0001 exception of when the inoculum concentration is at a low level of 20 mL,
concentration
AB 0.51 1 0.51 4.19 0.0800
where the COD removal initially increases from 66.87% to approxi­
AC 4.37 1 4.37 35.77 0.0006 mately 72.69% with increasing temperature before decreasing to
BC 0.029 1 0.029 0.24 0.6415 66.51%. Fig. 3 illustrates that the interactive effect of pH and inoculum
A2 139.51 1 139.51 1142.38 <0.0001 concentration on COD removal is insignificant while the temperature is
B2 103.85 1 103.85 850.33 <0.0001
35 ◦ C since the COD removal tends to change flatly with the concurrent
C2 89.36 1 89.63 733.91 <0.0001
Residual 0.85 7 0.12 – – increase in pH and inoculum concentration. It was obvious that inde­
Lack of Fit 0.056 3 0.019 0.094 0.9594 pendent changes in either pH or inoculum concentration have an equally
Pure Error 0.80 4 0.20 – – significant effect on COD removal when the other variable is a constant.
Corrected Total 409.81 16 – – – For instance, the increase in pH enhances COD removal from 66.38% to
Std. Dev. = 0.35, Mean = 71.16, C⋅V.% = 0.49, Press = 2.15, R2 = 0.9979, Adj. 68.85% at a 20 mL inoculum concentration and the increase in inoculum
R2 = 0.9952, Pred. R2 = 0.9948, Adeq. R2 = 45.519. concentration improves COD removal from 66.38% to 68.58% at pH 7.

3
J. Shi et al.
Interestingly, a comparison of COD removal by different factors
suggests that COD removal is reduced at a lower or higher temperature,
pH, or inoculum concentration, according to the consequences obtained
from the 3D surface curves. Jenny et al. (1991) first reported that Ba­
cillus licheniformis had the natural ability to produce a kind of lip­
opeptide biosurfactant. What’s more, surfactants are useful to form
thermodynamically stable oil-in-water emulsions in oily wastewater
(Posocco et al., 2016), enhancing the bioavailability of oily components
(Hwang et al., 2019). A lower or higher temperature has an adverse
influence on biosurfactant production and bacterial activity, resulting in
lower COD biodegradation efficiency. Additionally, pH is crucial to the
biodegradation ability of hydrocarbon-degrading bacteria (Thavasi
et al., 2007). Our experimental results exhibit that Bacillus licheniformis
is superior for COD removal in alkaline conditions than in neutral con­
ditions, which is compatible with the results reported by Khanpour-­
Alikelayeh et al. (2020). Furthermore, an optimal inoculum
concentration could effectively reduce the cost of the COD removal
process and the suitable concentrations for Bacillus licheniformis were in
the range of 1–5% (v/v) (Shajahan et al., 2017; Da Silva et al., 2021). On
the one hand, a higher inoculum concentration leads to a reduction in
biodegradation possibly due to insufficient environmental space and a
high mortality rate. On the other hand, a lower inoculum concentration
also causes less biodegradation mainly due to inadequate biomass.
3.4. Selected optimal conditions
Overall, the maximum COD removal attains 78.5941% in the
selected optimal conditions of 35.44 ◦ C, pH 8.13, and inoculum con­
centration of 31.47 mL. An experiment was implemented to validate the
predicted value, and the COD removal was found to be 77.81% in a SBR,
confirming that the predicted results fitted with the experimental re­
sults. In previous research regarding the COD biodegradation from bilge
water, Vyrides et al. (2018) demonstrated that the highest COD decrease
of 60% was observed by using the Moving Bed Biofilm Reactors with a
filling fraction of 40%. Further, Hwang et al. (2019) improved the COD
removal to 71.2 ± 0.3% in a single chamber microbial fuel cells with
anionic surfactant addition. By comparison, the findings of this study
have a clear advantage in terms of COD biodegradation. It is, however,
lower than 81% COD removal reported by Uma and Gandhimathi
(2019), who adopted a novel mixed bacterial consortium to degrade
4
Chemosphere 307 (2022) 135654
synthetic oily bilge water instead of real bilge water. As a result, making
use of a mixed bacterial consortium to promote COD removal from real
bilge water in a SBR would be a logical strategy for further study.
3.5. Practical performance of SBR in dealing with bilge water
The experimental diagram of SBR used in this study is presented in
Fig. 4. In comparison with the conventional oily water separator widely
equipped onboard, the SBR device has the characteristics of simple
process, stable operation effect, small footprint, and cost-saving, what’s
more, it is proud that it can purify the bilge water with the oil droplets
smaller than 20 μm, which cannot be achieved with oily water separator
equipment. SBR is an ideal batch-activated sludge treatment process,
which is currently in the stage of rapid development and improvement.
Although it has been widely verified in the treatment of industrial
wastewater and domestic sewage, the basic research and engineering
design of the bilge water treatment has just started, and more in-depth
research work is still needed.
The device utilized in this study is set up on the laboratory scale,
which has the shortcomings of low automation and small processing
capacity, but it has initially seen a good effect on the treatment of bilge
water. After improvement and promotion according to the actual
structure and needs of the ship, it is expected to be practically applied in
ships. Before that, it is currently necessary to fully evaluate the actual
processing effect of the device for future commercial scale-up.
To further investigate the implementation feasibility of SBR on ships,
the practical performance of the SBR in the treatment of bilge water is
necessary to explore under the optimal conditions of variables. SBR was
operated at an initial concentration of 8900 mg L− 1 and the results were
exhibited in Fig. 5 and Table S3. In its infancy, the HRT was maintained
at 5 days with an organic loading rate of 1.15 kg COD m− 3 d. During the
start-up stage of SBR, the degradation efficiency of COD is slow, because
it takes time for Bacillus licheniformis to acclimate to the environment
and acclimatization. After the SBR reached steady-state operation, the
maximum degradation rate of COD attained 77.8% after 12 days. The
OLR was varied by changing the HRT while keeping the initial con­
centration constant in order to examine its effect on COD degradation.
From day 13, the OLR was increased from 1.15 kg COD m− 3 d to 1.65 kg
COD m− 3d by reducing the HRT from 5 days to 4 days. The SBR was
monitored and observed that the maximum COD degradation rate of
J. Shi et al.
Fig. 1. Interactive effects of temperature and pH on COD removal.
74% was achieved after the SBR got steady-state within 7 days. After
that, the OLR was increased from 1.65 kg m− 3 d to 2.15 kg COD m− 3 d by
reducing the HRT from 4 days to 3 days on the 20th day. The results
obtained from this stage showed that the maximum degradation rate of
COD achieved 66.2% after 8 days of the SBR operation, which was a
significant decline compared with the COD degradation rate of the
previous stage.
From the results of Table S3, it is evident that the organic loading
rate increases instantaneously while decreasing the HRT, followed by a
decrease in the COD degradation rate. The difference is that when HRT
decreases from 4 days to 3 days, the decline in COD degradation rate is
far greater than that when HRT decreases from 5 days to 3 days. These
results indicate that SBR has an excellent ability to overcome some
interference caused by the change of organic loading rate, which was
well consistent with the outcomes of (Sathian et al., 2014) who imple­
ment experiments regarding decolorization and COD reduction of textile
dye wastewater in a SBR. They also found that with the decrease of HRT,
5
Chemosphere 307 (2022) 135654
the removal efficiency of COD decreased.
3.6. Biodegradation kinetic modeling
The biodegradation kinetic modeling is exceedingly crucial to opti­
mizing the biodegradation course (Sam et al., 2021), especially for the
practical microbiology prediction. In this study, a plot of COD concen­
tration remaining versus time under ideal operation conditions of SBR
after startup and stable operation for 6 days, as well as the fit of the first-
order kinetic equation to COD concentration remaining was depicted in
Fig. 6.
As shown in Fig. 6, the experimental values expressed were well
fitted to the first-order kinetic equation with a higher value of R2
0.98106 (R2 > 0.90). Finally, in a SBR, the obtained COD degradation
kinetic equation was written as equation (2):
Ct = 8900e− 0.10117t
(2)
J. Shi et al.
Fig. 2. Interactive effects of temperature and inoculum concentration on COD removal.
where, Ct is the COD concentration at t time (mg L− 1), t is the degra­
dation time (d), k is the biodegradation rate constant.
4. Conclusions
Using biodegradation to treat bilge water in SBR is a promising
method and may be applied to ships in the future. In this study, the
isolated Bacillus licheniformis was adopted for the biodegradation of
organic pollutants from bilge water in the SBR system. Meanwhile, the
process variables including temperature, pH and inoculum concentra­
tion were optimized by using RSM technology. Consequently, the
maximum COD removal value of 77.81% was achieved under the
optimal conditions. To further investigate the implementability of SBR
on ships, the practical performance of the SBR in the treatment of bilge
water is explored under the optimal conditions of variables, and the
results suggest that with the decrease of HRT, the degradation efficiency
6
Chemosphere 307 (2022) 135654
of COD decreased. Furthermore, the biodegradation process of COD was
well-suited to the first-order kinetic equation. The research data of this
study can provide some reference for treating bilge water by SBR.
Credit author statement
Jianqiang Shi: Methodology, Investigation, Formal analysis, Vali­
dation, Writing – original draft, Writing – review & editing. Guichen
Zhang: Resources, Investigation, Writing – review. Shaojun Zhang:
Resources, Data curation, Writing – review&editing, Validation. Run
Lu: Investigation, Writing – review & editing. Mengwei Chen: Investi­
gation, 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 influence
J. Shi et al. Chemosphere 307 (2022) 135654

Fig. 3. Interactive effects of pH and inoculum concentration on COD removal.

Fig. 4. Experimental diagram of sequential batch reactor.

7
J. Shi et al.
Fig. 5. Practical performance of SBR in COD degradation.
Fig. 6. COD concentration remaining curve and kinetic plot.
the work reported in this paper.
Data availability
No data was used for the research described in the article.
Acknowledgments
The work was funded by the National Natural Science Foundation of
China [grant number 51779136] and Science and Technology Com­
mission of Shanghai Municipality, China [grant number 20dz1206002].
The work was also supported by the Introduction and Education Plan of
Young Creative Talents in Universities of Shandong Province [grant
number 500076] and the Demonstration Base of Joint Cultivation of
Graduate Students in Shandong Province [grant number 2020–1].
Appendix A. Supplementary data
Supplementary data related to this article can be found at https://do
i.org/10.1016/j.chemosphere.2022.135654.
8
Chemosphere 307 (2022) 135654
References
Ahmad, N.A., Goh, P.S., Abdul Karim, Z., Ismail, A.F., 2018. Thin film composite
membrane for oily waste water treatment: recent advances and challenges. Membr.
Biochem. 8, 86.
Besharati Fard, M., Hamidi, D., Alavi, J., Jamshidian, R., Pendashteh, A., Mirbagheri, S.
A., 2021. Saline oily wastewater treatment using Lallemantia mucilage as a natural
coagulant: kinetic study, process optimization, and modeling. Ind. Crop. Prod. 163,
113326.
Church, J., Paynter, D.M., Lee, W.H., 2017. In situ characterization of oil-in-water
emulsions stabilized by surfactant and salt using microsensors. Langmuir 33,
9731–9739.
Da Silva, R.N., Melo, L.F.d.A., Luna Finkler, C.L., 2021. Optimization of the cultivation
conditions of Bacillus licheniformis BCLLNF-01 for cellulase production. Biotechnol.
Rep. 29, e00599.
Davarnejad, R., Mohammadi, M., Ismail, A.F., 2014. Petrochemical wastewater
treatment by electro-Fenton process using aluminum and iron electrodes: statistical
comparison. J. Water Proc. Eng. 3, 18–25.
Emadian, S.M., Hosseini, M., Rahimnejad, M., Shahavi, M.H., Khoshandam, B., 2015a.
Treatment of a low-strength bilge water of Caspian Sea ships by HUASB technique.
Ecol. Eng. 82, 272–275.
Emadian, S.M., Rahimnejad, M., Hosseini, M., Khoshandam, B., 2015b. Investigation on
up-flow anaerobic sludge fixed film (UASFF) reactor for treating low-strength bilge
water of Caspian Sea ships. J. Environ. Health Sci. 13, 23.
Eskandarloo, H., Selig, M.J., Abbaspourrad, A., 2018. In situ H2O2 generation for de-
emulsification of fine stable bilge water emulsions. Chem. Eng. J. 335, 434–442.
Gatidou, G., Samanides, C.G., Fountoulakis, M.S., Vyrides, I., 2022. Microbial electrolysis
cell coupled with anaerobic granular sludge: a novel technology for real bilge water
treatment. Chemosphere 296, 133988.
Ghelich, R., Jahannama, M.R., Abdizadeh, H., Torknik, F.S., Vaezi, M.R., 2019. Central
composite design (CCD)-Response surface methodology (RSM) of effective
electrospinning parameters on PVP-B-Hf hybrid nanofibrous composites for
synthesis of HfB2-based composite nanofibers. Compos. B Eng. 166, 527–541.
Gryta, M., 2020. Bilge water separation by membrane distillation. Separ. Purif. Technol.
37, 116332.
Guo, J., Wei, J., Huang, F., Massey, I.Y., Luo, J., Yang, F., 2021. Optimization of
microcystin biodegradation by bacterial community YFMCD4 using response surface
method. Chemosphere 274, 129897.
Hwang, J.-H., Kim, K.-Y., Resurreccion, E.P., Lee, W.H., 2019. Surfactant addition to
enhance bioavailability of bilge water in single chamber microbial fuel cells (MFCs).
J. Hazard Mater. 368, 732–738.
Jenny, K., KäPpeli, O., Fiechter, A., 1991. Biosurfactants from Bacillus licheniformis:
structural analysis and characterization. Appl. Microbiol. Biotechnol. 36, 5.
Kavitha, V., Mandal, A.B., Gnanamani, A., 2014. Microbial biosurfactant mediated
removal and/or solubilization of crude oil contamination from soil and aqueous
phase: an approach with Bacillus licheniformis MTCC 5514. Int. Biodeterior.
Biodegrad. 94, 24–30.
Khanpour-Alikelayeh, E., Partovinia, A., Talebi, A., Kermanian, H., 2020. Investigation of
Bacillus licheniformis in the biodegradation of Iranian heavy crude oil: a two-stage
sequential approach containing factor-screening and optimization. Ecotoxicol.
Environ. Saf. 205, 111103.
Li, P., Li, K., Xu, P., Liu, X., Pu, Y., 2021a. Treatment of wastewater with high carbon-to-
nitrogen ratio using a waterfall aeration biofilm reactor combined with sequencing
batch reactor: microbial community structure and metabolism analysis. Bioresource
Technol 337, 125450.
Li, Y., Zhang, M., Xu, D., Shan, X., Zheng, P., 2021b. Potential of anammox process
towards high-efficient nitrogen removal in wastewater treatment: theoretical
analysis and practical case with a SBR. Chemosphere 281, 130729.
Liu, Q., Niu, J., Yu, Y., Wang, C., Lu, S., Zhang, S., Lv, J., Peng, B., 2021. Production,
characterization and application of biosurfactant produced by Bacillus licheniformis
L20 for microbial enhanced oil recovery. J. Clean. Prod. 307, 127193.
Lu, D., Liu, Q., Zhao, Y., Liu, H., Ma, J., 2018. Treatment and energy utilization of oily
water via integrated ultrafiltration-forward osmosis–membrane distillation (UF-FO-
MD) system. J. Membr. Sci. 548, 275–287.
Magnusson, K., Jalkanen, J.-P., Johansson, L., Smailys, V., Telemo, P., Winnes, H., 2018.
Risk assessment of bilge water discharges in two Baltic shipping lanes. Mar. Pollut.
Bull. 126, 575–584.
Mazioti, A.A., Koutsokeras, L.E., Constantinides, G., Vyrides, I., 2021. Untapped
potential of moving bed biofilm reactors with different biocarrier types for bilge
water treatment: a laboratory-scale study. Water-Sui. 13, 1810.
Nayak, N.S., Purohit, M.S., Tipre, D.R., Dave, S.R., 2020. Biosurfactant production and
engine oil degradation by marine halotolerant Bacillus licheniformis LRK1. Biocatal.
Agric. Biotechnol. 29, 101808.
Pinheiro, D.R., Neves, R.d.F., Paz, S.P.A., 2021. A sequential Box-Behnken Design (BBD)
and Response Surface Methodology (RSM) to optimize SAPO-34 synthesis from
kaolin waste. Microporous Mesoporous Mater. 323, 111250.
Posocco, P., Perazzo, A., Preziosi, V., Laurini, E., Pricl, S., Guido, S., 2016. Interfacial
tension of oil/water emulsions with mixed non-ionic surfactants: comparison
between experiments and molecular simulations. RSC Adv. 6, 4723–4729.
Rice, E.W., Bridgewater, L., Association, A.P.H., Association, A.W.W., Federation, W.E.,
2012. Standard Methods for the Examination of Water and Wastewater. APHA.
Saber, A., Hasheminejad, H., Taebi, A., Ghaffari, G., 2014. Optimization of Fenton-based
treatment of petroleum refinery wastewater with scrap iron using response surface
methodology. Appl. Water Sci. 4, 283–290.
Sam, S.P., Tan, H.T., Sudesh, K., Adnan, R., Ting, A.S.Y., Ng, S.L., 2021. Phenol and p-
nitrophenol biodegradations by acclimated activated sludge: Influence of
J. Shi et al.
operational conditions on biodegradation kinetics and responding microbial
communities. J. Environ. Chem. Eng. 9, 105420.
Santiago, S.G., Morgan-Sagastume, J.M., Monroy, O., Moreno-Andrade, I., 2020.
Biohydrogen production from organic solid waste in a sequencing batch reactor: an
optimization of the hydraulic and solids retention time. Int. J. Hydrogen Energ 45,
25681–25688.
Sathian, S., Rajasimman, M., Radha, G., Shanmugapriya, V., Karthikeyan, C., 2014.
Performance of SBR for the treatment of textile dye wastewater: optimization and
kinetic studies. Alex. Eng. J. 53, 417–426.
Shajahan, S., Moorthy, I.G., Sivakumar, N., Selvakumar, G., 2017. Statistical modeling
and optimization of cellulase production by Bacillus licheniformis NCIM 5556
isolated from the hot spring, Maharashtra, India. J. King Saud Univ. Sci. 29,
302–310.
Soeprijanto, S., Nury, D.F., Pudjiastuti, L., 2019. Treatment of oily wastewaters from
tanjung perak port by electrocoagulation using aluminum electrodes. IOP Conf. Ser.
Mater. Sci. Eng. 543, 012098.
Tatar, D., 2021. Isolation, phylogenetic analysis and antimicrobial activity of halophilic
actinomycetes from different saline environments located near corum province.
Biologia 76, 773–780.
Chemosphere 307 (2022) 135654
Thavasi, R., Jayalakshmi, S., Balasubramanian, T., Banat, I.M., 2007. Effect of salinity,
temperature, pH and crude oil concentration on biodegradation of crude oil by
Pseudomonas aeruginosa. J. Biol. Environ. Sci. 1, 51–57.
Uma, V., Gandhimathi, R., 2019. Organic removal and synthesis of biopolymer from
synthetic oily bilge water using the novel mixed bacterial consortium. Bioresource.
Technol. 273, 169–176.
Uma, V., Gandhimathi, R., 2020. Effectiveness of ozone pretreatment on bioconversion
of oily bilge water into biopolymer. J. Water Process Eng. 36, 101275.
Vyrides, I., Drakou, E.-M., Ioannou, S., Michael, F., Gatidou, G., Stasinakis, A.S., 2018.
Biodegradation of bilge water: batch test under anaerobic and aerobic conditions
and performance of three pilot aerobic Moving Bed Biofilm Reactors (MBBRs) at
different filling fractions. J. Environ. Manage 217, 356–362.
Yang, F., Massey, I.Y., Guo, J., Yang, S., Pu, Y., Zeng, W., Tan, H., 2018. Microcystin-LR
degradation utilizing a novel effective indigenous bacterial community YFMCD1
from Lake Taihu. J. Toxicol. Environ. Health, Part A 81, 184–193.
Zhang, Y., Wei, H., Xin, Q., Wang, M., Wang, Q., Wang, Q., Cong, Y., 2016. Process
optimization for microcystin-LR degradation by Response Surface Methodology and
mechanism analysis in gas–liquid hybrid discharge system. J. Environ. Manage. 183,
726–732.