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Quantitative Evaluation of In-Situ Bioremediation of Compound Pollution of Oil and Heavy Metal in Sediments From The Bohai Sea, China
Quantitative Evaluation of In-Situ Bioremediation of Compound Pollution of Oil and Heavy Metal in Sediments From The Bohai Sea, China
A R T I C LE I N FO A B S T R A C T
Keywords: Owing to the semi-enclosed environment of the Bohai Sea, the ecological effects caused by an oil spill would be
In-situ bioremediation significant. A typical in- situ bioremediation engineering project for of oil-spilled marine sediments was per-
PAHs formed in the Bohai Sea and a quantitative assessment of the ecological restoration was performed. The bior-
Heavy mental emediation efficiencies of n-alkane and PAHs in the sediment are 32.84 ± 21.66% and 50.42 ± 17.49% after
Sediment
70 days of bioremediation, and 60.99 ± 10.14% and 68.01 ± 18.60% after 210 days, respectively. After
Evaluation
210 days of bioremediation, the degradation rates of two- to three ring PAHs and four-ring PAHs are
84.44 ± 23.03% and 26.62 ± 43.76%, respectively. In addition, the concentrations of the heavy metals first
increased by 6.00% due to oil spill degradation and release, and then decreased by 72.60% with the degradation
of oil caused by bioremediation or vertical migration. According to the continuous tracking monitoring, the
composition of the microbial community in the restored area was similar to that in the control area and the clean
area in Bohai Sea after 210 days of bioremediation. These results may provide some theoretical and scientific
data to understand the degradation mechanism and assessing the ecological remediation efficiency for oil spills
in open sea areas.
1. Introduction become the main pollutants in the Bohai Sea after nitrogen and phos-
phorus (SOAC, 2015). Polycyclic aromatic hydrocarbons (PAHs) are an
Driven by the development of agriculture and industry, as well as important class of persistent organic pollutants with carcinogenic,
the population explosion in China, the problem of marine environment mutagenic, and degradation-resistant properties that are of particular
pollution is getting increasingly serious (Wang et al., 2017). The Bohai concern (Yunker et al., 2002; Lima et al., 2005; Sofowote et al., 2008).
Sea (117°32′ E–122°08′ E, 37°07′ N–40°55′ N) is the primary area for Heavy metals are another kind of prevalent contaminant in the surface
ocean petroleum exploitation in China. Of the four offshore areas of sediments of the Bohai Sea. Unlike petroleum hydrocarbons, heavy
China, the oil platforms and submarine pipelines in the Bohai Sea are metals cannot be biologically or chemically degraded. In general, sev-
the most numerous and dense. The expansion of offshore oil and gas eral kinds of pollutants, such as PAHs and heavy metals, are always
exploitation has increased the risk of oil spill disasters. For example, an coexistent in the marine environment. The content of heavy metals in
oil spill accident occurred in the Penglai 19-3 oilfield of the central surface sediments in the central Bohai Sea is low (Xu et al., 2019),
Bohai Sea on June 11, 2011. The accident resulted in 840 km2 of pol- which meets the requirements of Class I standard for marine sediment
luted seawater area, which is worse than fourth-class according to the quality (Table 1; GB 18668-2002). In the typical Bay of Bohai Sea, Cd is
“Standards of water quality in the People's Republic of China (GB 3097- the most dangerous heavy metal followed by Hg, and the other five
1997)”. The highest oil content in the seawater was 1280 μg/l, which heavy metals, Pb, Cu, As, Cr, Zn, are of low ecological hazard (Xu et al.,
was 53-times higher than the background value. According to China 2017). At the polluted area of Penglai 19-3 oil spill accident, Pb, Cr, Ni,
Marine Environmental Quality Bulletin, the petroleum hydrocarbons Cu and Cd all reach the Class II standard of marine sediment quality,
⁎
Corresponding author at: Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, China.
E-mail address: cywang@yic.ac.cn (C. Wang).
https://doi.org/10.1016/j.marpolbul.2019.110787
Received 24 June 2019; Received in revised form 23 November 2019; Accepted 25 November 2019
0025-326X/ © 2019 Published by Elsevier Ltd.
Please cite this article as: Chuanyuan Wang, et al., Marine Pollution Bulletin, https://doi.org/10.1016/j.marpolbul.2019.110787
C. Wang, et al. Marine Pollution Bulletin xxx (xxxx) xxxx
Table 1
Concentration of heavy metal in sediment (mg/kg dw) and sediment quality guidelines of China for metal.
Item Hg Cd Pb Zn Cu Cr As
a
Standard for sediment quality Class I 0.20 0.50 60.0 150.0 35.0 80.0 20.0
Class II 0.50 1.50 130.0 350.0 100.0 150.0 65.0
Class III 1.00 5.00 250.0 600.0 2000 270.0 93.0
Bohai Seab Nd 0.06–0.22 8.7–32.3 28.7–61.2 6.7–34.6 Nd Nd
Bohai Seac Nd 0.23–0.44 42.8–73.6 Nd 38.1–61.9 89–219.1 20.6–253
2
C. Wang, et al. Marine Pollution Bulletin xxx (xxxx) xxxx
Fig. 2. Study area and the location of sampling sites in the Bohai Sea.
before the bioremediation engineering project was applied. Prior to authentic standards (nC10-40, Sigma). Alternately, individual PAHs were
conducting the ecological bioremediation, the surface sediment quantified based on the retention time and the m/z ratio of an authentic
(0–5 cm) was collected in August 2015 using an Ekman style stainless PAH mixed standard (Sigma). The relative abundance was calculated
steel grab sampler (0.1 M3). The location of the sampling station is from the respective mass chromatogram peak areas. Five surrogate
shown in Fig. 2. BH21 is non-oil and gas acquisition area and was used standards (naphthalene-d8, phenanthrene-d10, dibenzothiophene-d10,
as the control station without bioremediation. In this study, only some fluoranthene-d10, and pyrene-d10) were added to all the samples to
representative stations were located near oil platforms (the PL4, PL5, monitor the matrix effects. The average recoveries of surrogate stan-
PL6, PL11 and PL17 stations), and these were selected for quantitative dards varied from 81.6% to 92%. For the detailed analytical procedure,
evaluation of in-situ bioremediation based on a remediated region of conditions, and quality control refer to previous studies (Hu et al.,
0.76 km2. After 70 and 210 days of bioremediation, the surface sedi- 2011; Wang et al., 2017).
ment samples in the same stations near oil platforms and control area
were collected from the polluted area, and the oil content was de- 2.3.3. Instrumental analysis of heavy metals
termined to evaluate the ecological bioremediation efficiency. The samples were freeze-dried, crushed, and then sieved through a
100-mesh sieve. Approximately 0.1 g dried surface sediment samples
2.3. Analytical procedures were digested in 5 mL aqua regia (HF: HNO3: HClO4 = 5:2:1) in Toflon
cans using closed microwave digestion. They were then heated at
2.3.1. Extraction and fractionation 180 °C until the samples turned grey. Then 4 mL HNO3 was added into
Sediment samples were freeze-dried, extracted, and analyzed after the cooled samples. The residues were diluted with deionized water to
the biodegradation experiment. Detailed information on the sample 50 mL to determine the concentrations of the heavy metals. The con-
treatment and analysis had also been reported in previous studies (Hu tents of the heavy metals in the sample solutions were determined using
et al., 2011; Wang et al., 2011, 2017). Briefly, extraction was performed inductively coupled plasma mass spectrometry (ICP-MS; erkin Elmer
using the Soxhlet method with dichloromethane and methanol (93:7) as NexION 300Q, USA). The heavy metal concentrations were obtained
the solvent mixture for 72 h. The aliphatic hydrocarbons, aromatic using an average of three repeated measurements for each sample.
hydrocarbons and resins were separated by column chromatography
using activated alumina and silica gel, and gradient solvents as eluent: 2.4. Data analysis
ligarine, ligarine/dichloromethane (3:9, v/v) and MeOH, respectively.
Recoveries of crude oil groups varied from 89.54% to 93.75%. In ad- In this study, the average measurement value of the triplicate
dition, the recovery of crude oil components increased by 4.16% and samples found using GC-Ms and ICP-MS was used to develop figure and
was affected by the addition of Tween-20. table to show the regulation of data change. To evaluate the precision of
the analysis data, three replicates of each sample were tested every five
2.3.2. Instrumental analysis of the oil spill samples. Repeat values of the sample were presented as the mean ±
The instrumental analysis was performed on a Hewlett-Packard standard deviation (SD). The relative standard deviation (RSD) of the
6890 gas chromatograph interfaced with a Hewlett-Packard 5973 mass- replicates was 5.0%. For this sample, the average value was used for the
selective detector made by Agilent (USA). Helium was used as the convenience of drawing.
carrier gas. n-Alkane and PAHs were identified using the GC–MS in full The single factor index method was used to classify the water
scan mode. Sample extracts were injected in a splitless mode onto a H-5 quality by comparing the measured data with a standard, and to select
fused capillary column (30 m × 0.2 mm i.d) coated with 0.25 μm thick. the worst type of water quality as the evaluation result (Liu et al.,
Individual n-alkanes were identified based on the retention time of the 2017). This method was also used to evaluate the pollution status of
3
C. Wang, et al. Marine Pollution Bulletin xxx (xxxx) xxxx
petroleum hydrocarbons in the sediments in this study. The single 0d 70d 210d BR-70d BR-210d
3.1. Environmental quality assessment of petroleum hydrocarbons pollution 0d 70d 210d BR-70d BR-210d
(b)
The most abundant fraction in crude oil is alkanes, which are more
biodegradation, the bioremediation efficiency of n-alkane in the sedi-
likely to be utilized by microbes and easier to degrade (Nikolopoulou
ment were 32.84 ± 21.66% and 60.99 ± 10.14% after bioremedia-
et al., 2013; Wang et al., 2016). The oily sediment samples showed a
tion for 70 days and 210 days, respectively. Compared to long-chain n-
typical non-biodegraded profile, having a full suite of n-alkanes. How-
alkanes, compounds with low- to middle-molecular weight are gen-
ever, the samples (70 days and 210 days after bioremediation) showed
erally transformed and biodegraded at higher rates by the microbes.
significantly lower abundance of n-alkanes. The residual rate of total n-
Thus, the low molecular weight to high molecular weight n-alkane ratio
alkanes (T-ALK) in the sediment from the bioremediation area varied
(LMW/HMW) decreased greatly (Fig. 5a). Since the disappearance of
from 33.87% to 80.36% (avg. 62.11%, 70 days) and from 29.15% to
light components occurred primarily during the initial stage of de-
70.12% (avg. 39.00%, 210 days), respectively (Fig. 4a). However, the
gradation, the change in the LMW/HMW ratio further decreased, with
residual rate of n-alkane of the control station (BH21) without bior-
little magnitude after 210 days. In addition, the degree of oil biode-
emediation was very little. In the biodegraded oil residue, there was a
gradation was also assessed using the Pr/n-C17 (Didyk and Simoneit,
significant increase in the relative abundance of unresolved complex
1989; Lemkau et al., 2010). According to the already established pat-
mixtures (UCM). According to the analysis of hydrocarbon
tern, the higher Pr/n-C17 ratio for biodegraded oils (BR-70d, BR-210d)
than their non-biodegraded counterparts (0d) was observed (Fig. 5b).
Oil Platform
1000 45 Compared with the background survey, it was found that the con-
TPH(ppm)
4
C. Wang, et al. Marine Pollution Bulletin xxx (xxxx) xxxx
2.00 evaporation, but the loss of other PAHs was most likely due to biode-
0d gradation. The results further supported that the degree of the PAHs
BR-70d degradation decreased with an increasing in the number of aromatic
LMW/HMW ALK
3.4. Relationship between the heavy metal distribution and oil spill
(a) bioremediation
1.00
Heavy metals such as Cu, Zn, Ni, Cr, Fe, and Mn are essential mi-
0.50 cronutrients for various organisms including microorganisms, plants
and animals. The concentrations of heavy metals changed significantly
after remediation. From the variety of heavy metal concentrations, it
0.00 can be seen that the concentration of heavy metals increased from
600.13 mg/kg on a dry wt. basis (dw) to 635.70 mg/kg dw in the
BH21 PL17 PL5 PL6 PL11
bioremediation project after 70 days. Then a sharp decrease occurred
Station ID from 635.70 mg/kg dw to 174.15 mg/kg dw after 210 days of in-situ
bioremediation work. However, the Cu and Pb concentration showed a
1.50 continuous decrease with a relatively stable change (Fig. 7). Compared
0d with the environmental background value of the Bohai sediment (Li
BR-70d et al., 1994), the content of Pb, Cu and Cd in the area was higher than
BR-210d that of its background value prior to the bioremediation project. Among
1.00 them, concentrations of Pb were approximately three to five times the
Pr/n-C17
5
C. Wang, et al. Marine Pollution Bulletin xxx (xxxx) xxxx
Fig. 6. Relative degradation efficiency of PAHs with different numbers of rings with the bioremedition time.
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C. Wang, et al. Marine Pollution Bulletin xxx (xxxx) xxxx
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