Marine Pollution Bulletin 156 (2020) 111211

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Marine Pollution Bulletin

journal homepage: www.elsevier.com/locate/marpolbul

Membrane bioreactor and rapid sand filtration for the removal of T

microplastics in an urban wastewater treatment plant
⁎
Javier Bayo , Joaquín López-Castellanos, Sonia Olmos
Department of Chemical and Environmental Engineering, Technical University of Cartagena, Paseo Alfonso XIII 44, E-30203 Cartagena, Spain

A R T I C LE I N FO A B S T R A C T

Keywords: This paper discusses about the role of two different wastewater treatment technologies in the abatement of
Microplastics microplastics (MPs) from the final effluent of an urban wastewater treatment plant (WWTP); i.e., membrane
Wastewater treatment plant bioreactor technology (MBR) and rapid sand filtration (RSF). For this purpose, a WWTP with these two tech-
MBR nologies was monitored for 18 months. The average microplastic concentration was 4.40 ± 1.01 MP L−1 for the
RSF
influent, 0.92 ± 0.21 MP L−1 for MBR, and 1.08 ± 0.28 MP L−1 for RSF, without statistically significant
differences for MPs removal between both technologies (F-test = 0.195, p = 0.661). The main MP forms isolated
in our study were fibers (1.34 ± 0.23 items L−1), followed by films (0.59 ± 0.24 items L−1), fragments
(0.20 ± 0.09 items L−1), and beads (0.02 ± 0.01 items L−1). All of them probed to be statistically significant
reduced after both technologies, but without statistically significant differences between them. The MP removal
efficiency was 79.01% and 75.49% for MBR and RSF, respectively, although higher for microplastic particulate
forms (MPPs), 98.83% and 95.53%, than for fibers, 57.65% and 53.83% for MBR and RSF, respectively, dis-
playing a selective removal of particulate forms against microfibers. Fourteen different plastic polymers were
identified in the influent, only persisting low-density polyethylene (LDPE), nylon (NYL), and polyvinyl (PV) in
RSF effluent, and melamine (MUF) after MBR treatment. The MP size ranged from 210 μm, corresponding to NYL
fragment form in the influent, to 6.3 mm, corresponding to a red microfiber also from the influent. The max-
imum MP average size significantly decreased from MBR (1.39 ± 0.15 mm), to RSF (1.15 ± 0.08 mm) and
influent (1.05 ± 0.05 mm) (F-test = 4.014, p = 0.019), exhibiting the fiber selection carried out by these
advanced technologies for wastewater treatment.

1. Introduction
Microplastics as a global water pollutant was identified long time
ago, since Carpenter and Smith (1972) first described the presence of
tiny plastic particles floating in the Sargasso Sea, a region in the North
Atlantic Ocean without land boundaries, although the term “micro-
plastic” was first introduced in scientific references by Thompson et al.
(2004). MP pollution is a global threaten affecting, of course, the
marine environment, but also surface waters (Eriksen et al., 2013),
sediments (Ballent et al., 2016), air (Dris et al., 2016), soils (He et al.,
2018), food (Renzi et al., 2018) or drinking water (Barboza et al.,
2018). The sources of MPs include both fragmentation of larger plastic
items (secondary microplastics) and also direct input of micro-sized
plastic particles from personal care products or pre-production pellets
(Napper and Thompson, 2016).
Several sources have been pointed out as responsible for MPs in
WWTPs, both domestic and non-domestic ones. Domestic washing
machines are prone to release fibers from synthetic textiles. Browne
et al. (2011) reported a production of 1900 fibers per wash by a single
garment, Almroth et al. (2018) found average concentrations of 7360
fibers m−2 L−1 in polyester fleece fabrics and 110,000 fibers per gar-
ment and wash for PET fleece, and De Falco et al. (2018) reported an
average of 6,000,000 fibers from typical 5 kg load of polyester fabrics.
Besides MPs released from personal care products (Bayo et al., 2017),
paint scraps, pellets from plastic industries, manufactured fiber, tire
wreckage or microparticles from plastics consumer goods indeed reach
the sewage treatment, causing the WWTP to act as both a source and
sink for these micropollutants (Bayo et al., 2016; Bayo et al., 2020).
Wastewater is part of the urban water cycle, and its treatment is a
critical step that must be taken into account for the proper management
of such a scarce resource, especially in drought areas. For that reason,
the presence of MPs in wastewater effluents has gained more and more
attention during the last years (Talvitie et al., 2015; Michielssen et al.,
2016; Murphy et al., 2016; Ziajahromi et al., 2017; Simon et al., 2018;

⁎
Corresponding author.
E-mail address: javier.bayo@upct.es (J. Bayo).

https://doi.org/10.1016/j.marpolbul.2020.111211
Received 16 March 2020; Received in revised form 22 April 2020; Accepted 22 April 2020
Available online 04 May 2020
0025-326X/ © 2020 Elsevier Ltd. All rights reserved.
J. Bayo, et al.
Edo et al., 2020). Microplastics in WWTPs have been recognized only
by visual identification (Michielssen et al., 2016), with spectrometric
methods; i.e., Raman spectroscopy (Lares et al., 2018), FTIR (Bayo
et al., 2020), or with a combination of both methodologies, analyzing
all isolated microparticles (Bayo et al., 2020) or just a part of them
(Lares et al., 2018; Edo et al., 2020).
Laboratory experiments with MPs have shown various effects on
marine animals, such as swimming behavior (Choi et al., 2018), in-
creased mortality (Leung and Chan, 2018), reduced filtration rates,
DNA damage and neurotoxicity in bivalves (Ribeiro et al., 2017; Woods
et al., 2018) or malformations (Rist et al., 2019). Their potential tox-
icological effect, acting as vectors of both organic and inorganic ad-
ditives and chemicals (Bayo et al., 2018) are also important reasons for
their removal in the environment. This means an added risk for the
main predators, such as humans, that consume species contaminated
with microplastics or with chemicals released from these particles after
ingestion.
The efficiency of different technologies for the treatment of waste-
water effluents polluted with MPs has also been reported, although
some of them on a pilot scale or with short analysis periods, that could
not reveal actual seasonal variations in MP counts (Lares et al., 2018).
Membrane bioreactor technology (MBR) has proven to be an estab-
lished process for the treatment of both municipal and industrial was-
tewaters, with a high mixed-liqueur suspended solids concentration
that may benefit nitrifiers and other slow-growing microorganisms
(Dvořák et al., 2013). This technology has been tested for MP removal,
both in a real WWTP (Talvitie et al., 2017) or in a pilot-scale bioreactor
(Lares et al., 2018). On the other hand, rapid sand filtration (RSF) as a
tertiary treatment in WWTPs has also been reported as a wastewater
treatment technology for MP abatement. Hidayaturrahman and Lee
(2019) reported the smallest rate of MP removal by RSF when com-
pared with the use of ozone as a strong oxidant or a membrane disc-
filter. WWTPs are complex systems with chemical, physical and biolo-
gical processes taking place simultaneously, in order to achieve a high-
quality final effluent to be reused in agriculture or returned to the en-
vironment. In this paper we discuss about the role of two wastewater
treatment technologies; i.e., MBR and RSF, stages of a parallel process
differentiated and included in the same WWTP, located in Southeast
Spain.
2. Materials and methods
2.1. WWTP characteristics and sampling collection
The sewage treatment plant (EDAR Águilas) is a full-scale WWTP
treating both domestic and industrial wastewater, located in the Region
of Murcia (Southeast Spain) (37°25′29”N, 01°34′46”W), with a design
flow of 12,000 m3 d−1 and serving about 29,777 equivalent habitants
(Fig. S1). It consists of a double-flow plant, with an anoxic tank, a
biological reactor and a membrane filtration tank with a submerged
MBR unit. This membrane unit has a volume of 315 m3 with 10 flat-
sheet membrane units (EK-400, KUBOTA Corporation, Japan), com-
prising a total of 4000 membranes and an effective surface of 3560 m2.
The design flow is 1800 m3 d−1, and treated permeate water is directly
delivered to farmers for agricultural uses. The other flow is a conven-
tional activated sludge process with two biological reactors of a total
volume 3132 m3 and an advanced tertiary system consisting of three
rapid gravity sand filters installed in parallel. The hydraulic loading
rate is 6.68 m s−1, the effective grain size of the siliceous sand 0.9 mm,
the effective surface for each filter is 29.96 m2, and the allowed filtered
water for backwashing is 1.5% (Fig. S2). The average hydraulic reten-
tion time (HRT) for the conventional line is 17.28 h and 13.20 h for
winter and summer (July and August), respectively, and 17.76 h and
13.68 h for MBR, also for winter and summer (July and August), re-
spectively. Similarly, the sludge retention time (SRT) for the conven-
tional line is 9.75 d and 7.75 d for winter and summer (July and
2
Marine Pollution Bulletin 156 (2020) 111211
August), respectively, with similar values for MBR, although less dif-
ferences between winter and summer. A more detailed description of
both lines and the removal rates for the main water quality parameters
have been included as Supplementary Materials (Tables S1, S2, and S3).
Wastewater samples were collected at three stages of the treatment
process (Fig. S1); i.e., from the influent after grit and grease removal
(INF), in order not to block or clog filters and pumps with large debris,
after membrane bioreactor (MBR), and from tertiary treatment after the
rapid sand filtration (RSF). A total of 221.48 L of wastewater were
collected for this study, distributed in 73.45 L, 75.29 L, and 72.74 L for
INF, MBR and RSF, respectively. A total of 53 grab samples were pro-
cessed between February 14th 2018 and July 18th 2019, comprising 18
samples from INF, 17 from MBR, and 18 from RSF. Sample volumes
were precisely measured in each experiment, ranging from 3.20 to
5.37 L for INF (mean ± standard error) (4.08 ± 0.15 L), 3.40 to
9.20 L for MBR (4.43 ± 0.33 L), and 3.33 to 5.35 L for RSF
(4.04 ± 0.14 L), without statistically significant differences among
them (F-test = 0.921, p = .405). Samples were always grabbed in glass
bottles with metallic lid, and acquired in the morning, between 9 and
11 a.m. Samples from MBR and RSF were directly vacuum filtered
through a Büchner funnel using a paper filter (Prat Dumas, Couze-St-
Front, France, diameter 110 mm, pore size 0.45 μm) and MP contained
in wastewater samples from INF were previously isolated by means of a
density separation with a salt-saturated solution of NaCl, in a metho-
dology thoroughly reported in Bayo et al. (2020). All experiments were
carried out at room temperature (293 K).
The risk of contamination was also monitored. Only clothes made of
natural fabric, clean cotton lab coats and nitrile gloves were worn by
the analysts. The use of plastic lab devices was limited to the maximum,
although it could not be entirely avoided. All glassware was thoroughly
washed with tap water and twice with deionized water after each ex-
periment, covering it with aluminum foil to mitigate contamination. All
used containers were examined twice during the sampling campaign as
procedural blanks, by vacuum filtering 1.5 L of deionized water to
determine any potential microplastic contamination. No MP were iso-
lated from blank samples.
2.2. Microplastic analysis and dataset
2.2.1. Visual identification
Samples with possible MP were examined under a digital optical
trinocular microscope (Olympus SZ-61TR Zoom, Olympus Co., Tokyo,
Japan) coupled to a Leica MC190 HD digital camera and an image
capturing software Leica Application Suite (LAS) 4.8.0 (Leica
Microsystems Ltd., Heerbrugg, Switzerland), used for the analysis and
recording of color, shape and size of microparticles in their largest di-
mension. Once the images were captured, particles were successfully
isolated with tweezers in a 40-mm glass Petri dish for further testing by
FTIR. Microparticles were visually classified as: bead, a spherical piece;
fragment, an irregular shaped piece; fiber, a strand or filament piece;
and film, a thin sheet or membrane-like piece (Crawford and Quinn,
2017). We also followed a classification based on the size; i.e., mini-
microplastics (smaller than 1 mm) and microplastics (between 1 and
5 mm) (Crawford and Quinn, 2017). Eventually, microplastic forms
were also subdivided into microplastic fibers (MFBs), filament pieces
with a length at least five times its width, and microplastic particles
(MPPs) (Lares et al., 2018).
2.2.2. FTIR analysis
FTIR was used for the identification of functional groups and mo-
lecular composition of polymeric surfaces. Samples were compressed in
a diamond anvil compression cell, and spectra were acquired with a
Thermo Nicolet 5700 Fourier transformed infrared spectrometer
(Thermo Nicolet Analytical Instruments, Madison, WI, USA), provided
with a deuterated triglycine sulfate, DTGS, detector and KBr optics. The
spectra collected were an average of 20 scans with a resolution of
J. Bayo, et al.
16 cm−1 in the range of 400–4000 cm−1. Spectra were controlled and
evaluated by the OMNIC software without further manipulations, and
polymers were identified by means of different reference polymer li-
braries, containing spectra of all common polymers; i.e., Hummel
Polymer and Additives (2011 spectra), Polymer Additives and
Plasticizers (1799 spectra), Sprouse Scientific Systems Polymers by ATR
Library (500 spectra), and Rubber Compounding Materials (350
spectra). All statistical analyses were carried out with SPSS 26.0 sta-
tistic software (IBM Co. Ltd., USA). The critical value for statistical
significance was set at p < .050.
3. Results and discussion
3.1. General considerations
A total of 626 microparticles were isolated from all wastewater
samples, with an average concentration of 2.85 ± 0.49 items L−1, and
minimum and maximum values corresponding to 0 and 19.37 items
L−1 for INF, 0 and 2.67 items L−1 for MBR, and 0 and 5.28 items L−1
for RSF, with average concentrations of 5.71 ± 1.14 items L−1,
1.21 ± 0.22 items L−1, and 1.54 ± 0.35 items L−1 for INF, MBR, and
RSF, respectively (F-test = 12.486, p = .000). Every single isolated
particle was analyzed by FTIR, and 76.68% of these microparticles; i.e.,
480 items, were identified as microplastics, with an average con-
centration of 2.16 ± 0.42 MP L−1. This result emphasizes the im-
portance of additional spectroscopic approaches to successfully differ-
entiate MPs from other organic or inorganic non-plastic microparticles,
isolated during the extraction process. These mainly included calcium
stearate, glycerine and lipid mediators from solidified soap and other
washing liquids (Ziajahromi et al., 2017), as well as silicates, chipboard
fragments and animal fur. Gies et al. (2018) confirmed that just 32.4%
of the suspected MP were really plastic polymers, in samples from a
WWTP in Vancouver (Canada). Nevertheless, there is a difficult in
matching with high percentages because of the weathered and polluted
surfaces of microparticles. This was especially important for fibers
identification, where practical experience and criteria previously re-
ported were used (Song et al., 2015).
Although wastewater samples collected during 2019 proved to
display a higher average concentration of microplastics (2.84 ± 0.53
MP L−1) than samples collected during 2018 (1.71 ± 0.59 MP L−1),
there were no statistically significant differences (F-test = 1.785,
p = .187). We could not also verify statistically significant differences
by seasons (F-test = 0.865, p = .466). Statistically significant differ-
ences were observed between the average concentration of micro-
plastics collected in INF (4.40 ± 1.01 MP L−1) versus MP collected in
MBR (0.92 ± 0.21 MP L−1) and RSF (1.08 ± 0.28 MP L−1) (F-
test = 9.953, p = .000), indicating a clear removal of these micro-
pollutants with both advanced technologies. The average concentration
of microplastics in the incoming influent was lower than that detected
in major WWTPs located in Canada (31.1 ± 6.7 MP L−1) (Gies et al.,
2018) or in China (79.9 ± 9.3 MP L−1) (Liu et al., 2019);
(12.03 ± 1.29 MP L−1) (Yang et al., 2019), but higher than that re-
ported in a WWTP of our region with activated sludge process
(3.20 ± 0.67 MP L−1) (Bayo et al., 2020).
The removal percentage for MBR was 79.01%, higher than for RSF
with a 75.49%, although there were no statistically significant differ-
ences between both processes (F-test = 0.195, p = .661). These re-
moval rates were similar to that reported by Leslie et al. (2017) (72%)
in seven WWTPs located in Netherlands and by Liu et al. (2019)
(64.4%) in a sewage treatment plant in China, although lower than
other studies conducted in the United States (99.9%) (Carr et al., 2016),
United Kingdom (98.41%) (Murphy et al., 2016), Finland (98.3%)
(Lares et al., 2018) or China (95.16%) (Yang et al., 2019) (Table S4).
Although both MPPs and MFBs proved to decrease through the WWTP,
they displayed different removal rates. MPPs statistically significant
decreased from INF (2.26 ± 0.70 MPP L−1) to MBR (0.03 ± 0.02
3
Marine Pollution Bulletin 156 (2020) 111211
MPP L−1) and RSF (0.10 ± 0.05 MPP L−1) (F-test = 9.454, p = .000),
accounting for a 98.83% and 95.53% removal, respectively. In contrast,
changes in MFBs through the WWTP were smaller, from INF
(2.12 ± 0.55 MFB L−1) to MBR (0.90 ± 0.21 MFB L−1) (57.61%
removal) and RSF (0.98 ± 0.27 MFB L−1) (53.83% removal), still
statistically significant (F-test = 3.214, p = .049). Lares et al. (2018)
also reported different removal efficiencies for MPPs (89.9%) than for
MFBs (99.1%), and Talvitie et al. (2017), although with better removal
efficiencies, only detected two synthetic textile fibers after the MBR
technology.
In this study, although it is clear that a good performance of MBR
and RSF for MPPs removal is achieved, MFBs still bypass MBR and
escape into the aquatic environment, particularly because of the high
pressure applied in this system (Leslie et al., 2017), giving a global
lower yield than a conventional activated sludge process (Bayo et al.,
2020). A direct air pollution with fibers derived from apparel articles
and household dust can also be released in open-air sewage treatment
plants (Zobkov and Esiukova, 2017). Besides, the small size and mor-
phology of fibers also enables them to longitudinally pass through the
RSF (Talvitie et al., 2017; Ziajahromi et al., 2017).
3.2. Distribution of MPs in wastewater samples
3.2.1. Size, shape and color distribution
Four shapes were identified in all wastewater samples; i.e., fibers
(1.34 ± 0.23 items L−1, 61.09%), films (0.59 ± 0.24 items L−1,
31.5%), fragments (0.20 ± 0.09 items L−1, 6.7%), and beads
(0.02 ± 0.01 items L−1, 0.6%), clearly showing that fibers were the
most abundant form type isolated in this study. These results proved to
be different to those previously reported in a WWTP from our Region,
with a conventional activated sludge process, where fragments were the
most abundant type of microplastics identified (46.9%) (Bayo et al.,
2020), but similar to that described by Lares et al. (2018) in a pilot-
scale membrane bioreactor operated in a municipal WWTP. In fact, the
ratio of MFBs and MPPs versus total microplastics was 48.09% and
51.91% in the INF, increasing after the MBR (96.72% and 3.28%) and
the RSF (90.79% and 9.21%), respectively.
Fibers were the dominant form type in all seasons, decreasing
during the Summer (0.94 ± 0.24 items L−1) with respect to Winter
(1.69 ± 0.63 items L−1), Autumn (1.58 ± 0.56 items L−1) and Spring
(1.21 ± 0.36 items L−1), although without statistically significant
differences (F-test = 0.516, p = .673). The release of important
amounts of synthetic fibers from washing machines, depending on
textile properties, washing conditions, type of detergent, softener and
garment weathering, has been previously reported (Browne et al., 2011;
Almroth et al., 2018; De Falco et al., 2018; Prata, 2018). Moreover,
fibers were by far the most common type in both size groups, mini-
microplastics (< 1 mm) and microplastics (1–5 mm), accounting for
54.2% and 70.7%, respectively. Most fragments were categorized as
opaque (71.88%), and most films as transparent (70.86%). These re-
sults are similar to that reported by Leslie et al. (2017) in WWTPs from
The Netherlands. In the case of film form, the average concentration
decreased from INF (1.70 ± 0.64 items L−1) to RSF (0.06 ± 0.03
items L−1), totally disappearing in MBR effluent (F-test = 6.596,
p = .003).
According to the size, 58.90% of MPs isolated from all samples were
smaller than 1 mm, being 30.72% between 1 and 2 mm. The minimum
detected size was 210 μm for a nylon fragment isolated in INF. As de-
picted in Fig. 1, the percentage of each size was different depending on
the stage in the WWTP. The main size range in all wastewater samples
was between 1 and 2 mm; i.e., 24.33% for INF, 37.29% for MBR and
36.84% for RSF. There was no stage with microplastics lower than
200 μm. Average microplastic size reported a statistically significant
increase from INF (1.05 ± 0.05 mm), to RSF (1.15 ± 0.08 mm) and
MBR (1.39 ± 0.15 mm) (F-test = 4.014, p = .019) indicating the fiber
selection made by advanced treatment technologies previously
J. Bayo, et al.
Fig. 1. Size classification at three monitored sampling points, according to Spanish Environmental Ministry.
discussed.
A total of 12 different colors were detected in all samples, sug-
gesting a number of different sources for these microplastics Fig. 2. Blue
was the most common color for MFBs (83.80%), while MPPs were
mostly white or uncolored (56.99%). These results are similar to those
reported by our research group in another WWTP (Bayo et al., 2020).
Colored microplastics are the most dominant in water samples, ac-
counting for 50.4–86.9% (Chen et al., 2020). In our study, 76.15% were
colored ones, being blue, grey, green, yellow, brown, orange, pink and
red the most common colors (Fig. 4). The obvious presence of dyed
fibers is also a well-known fact in wastewater samples (Edo et al.,
2020). The ability of predators for ingesting microplastics with colors
resembling their prey has been thoroughly described (Boerger et al.,
2010), exhibiting different inhibition effects according to the color
(Chen et al., 2020).
3.2.2. Polymer types
A total of 14 different polymer types were identified in all waste-
water samples, as presented in Table 1, being low-density polyethylene
(LDPE) the most common one (70.61%), with an average concentration
of 0.59 ± 0.22 LDPE L−1 and a maximum value of 10.80 LDPE L−1 in
an INF sample. This polymer family was followed by high-density
polyethylene (HDPE) (5.16%), acrylate (AC) (4.97%), polypropylene
(PP) (4.95%), polystyrene (PS) (4.02%), nylon (NYL) (3.01%), metha-
crylate (MCR) (1.67%), polyvinyl (PV) (1.63%), poly(ethylene:propy-
lene) (EPM) (1.01%), and melamine (MUF) (1.01%), the last one only
isolated in a MBR wastewater sample. Melamine is recalcitrant and
toxic to bacteria in conventional activated sludge systems, although low
mass ratio of toxic melamine to biomass in the MBR proved not to affect
its performance (Xu et al., 2018). Samples from RSF revealed the pre-
sence of LDPE (2.47%), NYL (0.58%), and PV (1.16%).
Mintenig et al. (2017) also reported 14 different polymers, poly-
ethylene being the most common one, and Akarsu et al. (2020) found
polyethylene in 51% of plastic particles obtained from the effluent of
three WWTPs in Turkey, similar to that reported by our research group
in another WWTP with a conventional sludge process (52.4%) (Bayo
4
Marine Pollution Bulletin 156 (2020) 111211
et al., 2020). As presented in Fig. 3a, LDPE compounds have distinct
absorption bands for CH and CH2 groups between 2930 and 2850 cm−1
wavenumbers, due to an asymmetric vibration of CH2,
1470–1450 cm−1 for bending CeC bond between methylene carbons,
and 730–720 cm−1 band due to rocking in plane of CH2 groups (Castillo
et al., 2016). Analysis of variance indicated that there were statistically
significant differences in LDPE from the INF (1.69 ± 0.59 LDPE L−1)
to the RSF (0.06 ± 0.04 LDPE L−1) (F-test = 7.634, p = .001), totally
disappearing after the MBR.
Several acrylate polymers were also identified in wastewater sam-
ples; i.e., poly(lauryl acrylate) (Fig. 3b), poly(cyclohexyl acrylate), or
poly(11-bromoundecyl acrylate), among others. They are widely used
in many daily applications, including commercial shower gels, peelings,
waterproof sunscreen, or as a gelling agent in lipsticks and paint par-
ticles (Liebezeit and Dubaish, 2012; Chae et al., 2015), and all of them
were removed after MBR or RSF.
Polypropylene (Fig. 3c) has absorption bands at 2950–2838 cm−1
for CeH stretch, 1455–1377 cm−1 for CH2 bend, and 997–972 cm−1 for
CH3 rock, among others (Jung et al., 2018). The weak spectral band
around 1650 cm−1 (marked with an arrow) identifies the carbonyl
group (C=O), indicating, to some extent, oxidation of PP favored by
our warm and arid climate, as previously reported by Castillo et al.
(2016) in samples from Qatar. Microplastics of PP totally disappeared
after MBR and RSF (F-test = 12.657, p = .000). Polystyrene (dicarboxy
terminated) (Fig. 4a) showed vibration modes between 3050 and
2850 cm−1 because the aromatic ring = CH stretch, 1600–1500 cm−1
for the aromatic C]C stretch, and the ring band at 750–600 cm−1
(Chae et al., 2015; Jung et al., 2018). As for polypropylene, PS micro-
plastics were completely removed after both wastewater treatments;
i.e., MBR and RSF (F-test = 3.454, p = .039). Other polymer types,
reported in relatively high number in beach sediments, because is its
use in shipbuilding (PV) and packaging (EPM), only represented 1.63%
and 1.01% in wastewater samples, respectively. Styrene-butadiene
rubber (SBR) (Fig. 4c) was hardly isolated (0.009 items L−1), despite
being a potential marker for the identification of tread wear particles
from car-tire abrasion. It is a high density compound (between 1.7 and
J. Bayo, et al. Marine Pollution Bulletin 156 (2020) 111211

Fig. 2. Microplastics in different stages of the WWTPs identified by FTIR: (a) low density polyethylene fragment (LDPE) (RSF/14th.Feb.2018); (b) polypropylene
fiber (PP) (MBR/14th.Feb.2018); (c) polystyrene bead (PS) (INF/14th.Feb.2018); (d) polystyrene fragment (INF/14th.Feb.2018); (e) Acrylate film (ACRYL) (INF/
14th.Feb.2018); (f) unknown fiber (MBR/10th.May.2018); (g) melamine fragment (MUF) (MBR/7th.Jun.2018); (h) polyamide film (PA) (RSF/1st.Aug.2018); (i) low
density polyethylene (LDPE) (RSF/13th.Sep.2018); (j) nylon (NYL) (INF/13th.Sep.2018); (k) polyester (PEST) (INF/11th.Oct.2018); (l) styrene-butadiene copolymer
(SBR) (INF/18th.Jul.2019).

Table 1 associated microplastic particles is expected to be found in the runoff

Polymer types detected in wastewater samples from EDAR Águilas (INF, MBR from the road and road verge generated during rainfall events. Floods
and RSF), expressed as percentage (%) of MPs concentration. produced after extreme hydrological events in the Mediterranean area
Polymer families INF (%) MBR (%) RSF (%) Total (%) cause the sweeping of these microplastics and their immediate exit from
the treatment plant, making detection also difficult.
Acrylate (AC) 4.97 – – 4.97 The other polymer types found in wastewater sample were biopo-
Biopolymer (BPL) 0.59 – – 0.59
lymer (BPL) (0.59%), polyester (PEST) (0.53%), teflon (PTFE) (0.42%),
High-density polyethylene (HDPE) 5.16 – – 5.16
Low-density polyethylene (LDPE) 68.14 – 2.47 70.61 and polyisobutylene (PIB) (0.42%), being the last one a rarely identified
Melamine (MUF) – 1.01 – 1.01 plastic type, also reported in oysters from the French Atlantic coast
Methacrylate (MCR) 1.67 – – 1.67 (Phuong et al., 2018).
Nylon (NYL) 2.43 – 0.58 3.01
Poly(ethylene:propylene) (EPM) 1.01 – – 1.01
Polyester (PEST) 0.53 – – 0.53
4. Conclusions
Polyisobutylene (PIB) 0.42 – – 0.42
Polypropylene (PP) 4.95 – – 4.95 The present study showed that microplastics represented a 76.68%
Polystyrene (PS) 4.02 – – 4.02 of total microparticles isolated in wastewater samples from the INF,
Polyvinyl (PV) 0.47 – 1.16 1.63
MBR and RSF, emphasizing the need for a specific discrimination
Teflon (PTFE) 0.42 – – 0.42
technique for their identification. The removal efficiency was 79.01%
for MBR and 75.49% for RSF, not confirming that wastewater advanced
2.5 g cm−3) and difficult to be quantified by FTIR due to the carbon technologies are better to retain microplastics than conventional one;
black added as a filler (Haave et al., 2019). Besides this fact, rainfall i.e., activated sludge processes. Fibers were the most abundant type,
scarcity is a distinctive climate characteristic of this area and, as re- increasing from INF (48.09%) to RSF (90.79%) and MBR (96.72%).
ported by Vogelsang et al. (2019), the major fraction of road dust- They were also the dominant form type in all seasons, decreasing from
Summer to Winter, although without statistically significant

5
J. Bayo, et al. Marine Pollution Bulletin 156 (2020) 111211

95 (a)
Microplastic
90 (match: 95.15%)
85
80 Reference LDPE
75
70
65
60
%Transmittance

55
50
45
40
35
30
25
20
15
10
4000 3500 3000 2500 2000 1500 1000 500
Wavenumbers (cm-1 )

Reference AC
95
(b)
90
85
80
75
70
65 Microplastic
%Transmittance

60 (match: 79.13%)
55
50
45
40
35
30
25
20
15
10
4000 3500 3000 2500 2000 1500 1000 500
Wavenumbers (cm-1 )

Reference PP
95
(c)
90
Microplastic
85 (match: 91.34%)
80
75
70
65
%Transmittance

60
55
50
45
40
35
30
25
20
15
10
4000 3500 3000 2500 2000 1500 1000 500
Wavenumbers (cm-1 )

Fig. 3. FTIR spectra with references polymers and microplastics: (a) Fragment of poly(ethylene) (low density) INF 26th.Mar.2019 (95.15% match) (Sprouse Scientific
Systems Polymers by ATR Library); (b) Film of poly(lauryl acrylate) INF 11th.Apr.2018 (79.13% match) (Sprouse Scientific Polymers by ATR Library). (c) Film of
polypropylene, isotactic INF 5th.Jul.2018 (91.34% match) (Hummel Polymer and Additives).

differences. Average microplastic size increased from throughout the (4.95%).

WWTP, being 58.90% of microplastics smaller than 1 mm.
Microparticles were made of 14 different polymer types, with the ma-
jority identified as LDPE (70.61%), HDPE (5.16%), AC (4.97%), and PP

6
J. Bayo, et al. Marine Pollution Bulletin 156 (2020) 111211

Reference PS
95
90 (a)
85
Microplastic
80 (match: 93.40%)
75
70
65
%Transmittance

60
55
50
45
40
35
30
25
20
15
10
4000 3500 3000 2500 2000 1500 1000 500
Wavenumbers (cm-1 )

102 Reference EPM

100
98
(b)
96 Microplastic
(match: 86.09%)
94
92
90
88
%Transmittance

86
84
82
80
78
76
74
72
70
68
4000 3500 3000 2500 2000 1500 1000 500
Wavenumbers (cm-1 )

Reference SBR
95
90 (c)
85 Microplastic
80 (match: 82.15%)

75
70
65
%Transmittance

60
55
50
45
40
35
30
25
20
15
10
4000 3500 3000 2500 2000 1500 1000 500
Wavenumbers (cm-1 )

Fig. 4. FTIR spectra with references polymers and microplastics: (a) Fragment of poly(styrene), dicarboxy terminated INF 14th.Feb.2018 (93.40% match) (Sprouse
Scientific Polymers by ATR Library); (b) Film of poly(ethylene:propylene) (60% ethylene) INF 14th.Feb.2019 (86.09% match) (Sprouse Scientific Polymers by ATR
Library); (c) Fragment of styrene-butadiene copolymer INF 18th.Jul.2019 (82.15% match) (Polymer Additives and Plasticizers).

7
J. Bayo, et al.
CRediT authorship contribution statement
Javier Bayo:Methodology, Formal analysis, Writing - original
draft, Investigation, Data curation, Visualization.Joaquín López-
Castellanos:Conceptualization, Methodology, Supervision,
Funding acquisition, Investigation, Resources.Sonia
Olmos:Investigation, Resources, Supervision, Visualization.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgements
This work was financed by Project 5245/18IQA (Cetenma and
Hidrogea). Analyses carried out by S. Olmos were supported by a grant
from Fundación Séneca (20268/FPI/17). Authors gratefully acknowl-
edge the work and cooperation of personnel of the WWTP of Cartagena
with wastewater samples collection.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.marpolbul.2020.111211.
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